Argyrophilic nucleolar organizer regions in breast cancer: prognostic significance. 109 cases of breast cancer were examined in order to assess correlation between the number and size of argyrophilic nucleolar organizer regions (AgNORs) on one hand and prognosis of the disease on the other hand. Results indicate that an indirect correlation exists between AgNOR counts and survival rate of patients. Cases with short survival were observed having significantly frequent AgNORs larger than 5 microns.


[Combined small cell and "adenoid cystic" carcinoma of the uterine cervix]. Cervical carcinoma in a 49-year-old woman was resected and found to spread into parametrium. It consisted of a unique combination of adenoid cystic, squamous and small cell neuroendocrine structures. Small celled part revealed positivity with chromogranin A and neuron-specific enolase, neuroendocrine granules were argyrophilic and about 130 nm in diameter. Adenoid cystic and squamous parts were lacking of unambiguous neuroendocrine or myoepithelial differentiation. The patient died in 27 months due to generalization but only a small cell carcinoma without any heterogeneity was found in autopsy. Small cell and adenoid cystic carcinomas are rare and more malignant than usual cervical carcinomas.


Zinc triflate-promoted glycosidation: synthesis of lipid A disaccharide intermediates. Zinc triflate was found to be superior to the heavy metal salts as a promoter in the Koenigs-Knorr type glycosidation reaction in the synthesis of lipid A disaccharide intermediates. It readily promoted the reaction of a complex glycosyl bromide with a reducing sugar moiety and gave the disaccharide with beta-selectivity in good yield. This method would be suitable for the bulk preparation of lipid A disaccharide intermediates.


Malignant biliary obstruction: treatment with self-expandable stainless steel endoprosthesis. Metal endoprostheses of the Wallstent type were successfully inserted percutaneously and endoscopically in 80 consecutive patients with malignant obstructive biliary stenoses, who were followed for up to 18 months. The indication for treatment was jaundice due to malignant biliary obstruction. Repeat radiological investigations were performed if the patient had symptoms suggesting stent occlusion. After stent implantation, 88% of patients demonstrated a serum bilirubin decrease by more than 50%. We observed a 15% rate of serious complications, including a 10% rate of cholangitis with septicemia. There were no cases of stent migration or occlusion due to encrustation of bile. Recurrent jaundice occurred in 17.5% of patients due to progressive tumor growth after 3-10 months. In 5 of these patients, tumor overgrowth was redilated and/or restented. Of the 80 patients, 34% are alive after 2-12 months (mean: 242 days); of these, two-thirds are free of jaundice. Sixty-six percent of patients died between 3 days and 1.5 years (mean: 133 days). Although autopsy investigations revealed the possibility of tumor growth onto the inner surface of the stent, through the mesh of the endoprosthesis, no stent occlusion by tumor ingrowth into the lumen occurred. Self-expandable stainless steel endoprostheses provide good palliation in patients with malignant obstructive jaundice.


[Immunohistochemical and histochemical study on colorectal adenomas and polyps]. Immunohistochemical staining and histochemical approach for mucin were applied in studying 58 cases of colorectal adenomas. Among them, there were 13 cases of multiple adenomas (polyps), 5 cases of inflammatory polyps and 5 cases of juvenile polyps. The results indicated that positive expression of both McAb MC5 and Ulex europaeus agglutinin-1(UEA-1) were correlating with the degree of dysplasia, the histological type and the size of adenomas. The positive expression of peanut agglutinin (PNA) was correlating with the degree of dysplasia. Moderate and heavy stainings were mainly seen in adenomas accompanying with moderate and severe dysplasia, as well as adenomas with early carcinomatous changes. PAT-KOH-PAS method could sensitively reflect the occurrence of dysplasia and malignancy of the adenomas. These findings support the concept of an adenoma(dysplasia)-carcinoma sequence. In comparing multiple adenomas (number of adenomas > 100) with the solitary ones, a notable difference was obtained in the expression of McAb MC5. It's concluded that combined use of certain immunohistochemical and histochemical stainings is well be useful in detecting the malignant potentiality of adenoma.


Inhibitory effect of bis(2-aminohexyl) disulfide and bis(2-amino-3-phenylpropyl) disulfide on several mouse inflammations. The anti-inflammatory profile of the analogues of bis(2-aminopropyl) disulfide dihydrochloride with butyl (compd. II) and phenyl (compd. III) instead of the methyl group was studied in several mouse models related to phagocyte functions. The test samples were administered 2-3 h before the inflammatory stimulation or the peak of inflammation. Subcutaneously administered, compds. II and III significantly inhibited serotonin-induced paw edema in a dose-dependent manner (50% inhibitory dose values: 10 and 5 mg/kg, respectively), when orally administered at 25 mg/kg, these compounds were significantly effective, but their potencies were weaker. Neither compound had any irritant activity when administered at a dose of 12.5 micrograms/5 microliters/paw into the paw. In a sheep red blood cells (SRBC)-induced delayed-type hypersensitivity (DTH) reaction model, compd. II (25 mg/kg, s.c.) significantly inhibited the DTH responses when administered at two different times in relation to the time of challenge. However, there was only slight inhibition by compd. III (25 mg/kg, s.c.) on paw edema formation when administered 14 h after secondary immune response. In a model of experimental acute hepatic failure induced by successive injections of Propionibacterium acnes and lipopolysaccharide, both compounds increased mouse survived, compared with the control mice, and kept the serum levels of components involved in hepatic failure to nearly normal levels. These results demonstrate that compds. II and III possess an inhibitory effect on inflammation related to phagocytes.


Metallic stents for the treatment of biliary obstruction: a report of 100 cases. The results of the first 100 patients to receive Gianturco-Rösch "Z" stents is presented along with recommendations for their use. The patient population was comprised of 57 men and 43 women, age range 17-85 years (mean 65 years). Fifty-four of the patients had benign obstruction and 46 had malignant obstruction. Of the benign lesions, 11 had sclerosing cholangitis and the remainder had postoperative strictures. Thirty-one of the malignant obstructions were secondary to cholangiocarcinoma with the majority of the others secondary to metastases from various sources. All but one had multiple systems involved. Patients with benign postoperative strictures were all initially treated with balloon angioplasty; if this failed, stents were inserted. In patients who had stents in place for greater than 1 year, the occlusion rate was 13%. The overall occlusion rate in the 43 patients was 7%. Patients with sclerosing cholangitis did less well. In those with sclerosing cholangitis secondary to intraarterial chemotherapy, the occlusion rate was 77%, and we no longer use the metallic "Z" stent in these patients. The stent was not used for malignant common duct obstruction. All patients had hilar involvement. In the patients with malignant obstruction, 17% re-obstructed prior to their death. The patients with cholangiocarcinoma did well with a mean survival time of 14 months and a re-obstruction rate of 16%. All late obstructions were secondary to tumor overgrowth either proximal or distal to the stents. We conclude that the "Z" stent is an effective form of treatment in patients with benign postoperative strictures and those with malignant obstruction involving the hilum. We do not recommend it as a replacement for conventional stents, but rather as an additional device that allows treatment of some of the more difficult causes of obstruction.


[An immunohistochemical and ultrastructural study of 243 gliomas with reference to the prognosis]. The localization of S-100 protein, glial fibrillary acidic protein (GFAP) and vimentin as well as the relations between immunohistochemical reactions, grading and prognoses in 243 gliomas were investigated. In astrocytomas, both GFAP and S-100 protein were decreased along with the increase of tumor grades while the vimentin content was coincidently increased. The intensity of GFAP were negatively related to that of vimentin. Typical neoplastic oligodendroglial cells were known to be devoid of glial filaments and negative to GFAP and vimentin, anyhow, in the sporadic tumor cells, positive reaction to both GFAP and vimentin were identified, and these cells were considered to be either the astrocytes or the transitional cells between astrocytes and oligodendrocytes. Difference of the survival rates in cases with various gradings or intensities against S-100 protein and GFAP stainings were noticed. The results suggested that vimentin and GFAP may exist either independently or coexist synchronously in the astrocytomas as the markers, expressing the anaplasia stages of astrocytes. S-100 protein is considered to be an important indicator for both differentiation and prognosis of this tumor.


[Epithelial metaplasia in gastric or intestinal type and malignant transformation in mucinous ovarian tumors]. By using HE, histochemistry and immunohistochemistry stainings, 64 cases of mucinous tumors of ovaries including 24 benign, 29 borderline and 11 malignant cases were studied. The epithelial lining was divided into 4 types, namely: cervical, intermediate, gastric and intestinal types. According to the morphological and histochemical characteristics the intermediate type epithelium might be a transitional form between the cervical type and the gastric or intestinal metaplastic type. Argyrophil cells appeared only in the gastric or intestinal type epithelium, and malignant transformation was also to be detected in these 2 types. The constituents and amounts of mucin showed obvious difference among these four types of epithelium. The amount of neutral mucin was prominently decreased during the process of malignant transformation of the intestinal type epithelium and the expression of sulphomucin was also slightly predominant than the sialo-mucin's, but these changes were not noticed during the malignant change of gastric type epithelium.


Reversible kinase and phosphatase regulation of brain amino acid receptors in postnatal development. In a previous study [Shaw, C., Pasqualotto, B. and Lanius, R.A., Mol. Neuropharmacol., in press] we have shown that phosphorylation and dephosphorylation actions of protein kinase and alkaline phosphatase lead to decreases or increases in the number of GABAA and AMPA receptors in adult rat neocortex. Using the same in vitro cortical slice preparation, we have now examined the role of these enzymes in regulating GABAA and AMPA receptors at different stages of postnatal development. GABAA receptors were labelled with [3H]SR95531 [Shaw, C. and Scarth, B.A., Mol. Brain Res., 11 (1991) 273-282]; AMPA receptors were labelled with [3H]CNQX [Lanius, R.A. and Shaw, C., Mol. Brain Res., 15 (1992) 256-262]. At postnatal day 14, GABAA receptors showed a decrease in binding in response to alkaline phosphatase treatment as opposed to an increase in binding observed in response to protein kinase treatment. Similar effects were observed for AMPA receptors at 20 days of age. The direction of regulation following the enzyme treatments were opposite to those observed in the adult cortex for both receptor populations. These fundamental changes in the enzymatic nature of regulation for such key inhibitory and excitatory receptor populations in cortex may signal an important role for age-dependent kinases and phosphatases in the events leading to modifications in neuronal function during postnatal development.


Morphine regulates DNA synthesis in rat cerebellar neuroblasts in vitro. The effects of morphine on DNA synthesis by external granular layer (EGL) neuroblasts was examined in whole-mount organotypic cultures isolated from 10-day-old rat cerebella using bromodeoxyuridine (BrdU). After 24 h in vitro, explants were treated for 24 h with 10 nM, 1 or 100 microM morphine, morphine plus 30 nM, 3 or 300 microM of the opiate antagonist naloxone, respectively, or those concentrations of naloxone alone. BrdU was added during the last 4 h of drug treatment. EGL neuroblasts were unambiguously identified by size and morphology, location and by protein kinase C II immunocytochemistry. The proportion of EGL neuroblasts incorporating BrdU was significantly reduced in the presence of 1 microM morphine, while 100 microM morphine had little additional effect. The concentration of morphine predicted to cause a half-maximal reduction in BrdU labeling index was 22.5 nM. Morphine's ability to reduce BrdU incorporation by EGL neuroblasts was concentration dependent and was prevented by concomitant treatment with naloxone, implicating the involvement of opioid receptors. The results suggest that morphine can directly regulate the growth of the developing cerebellum by inhibiting neuroblast proliferation within the EGL.


An enzymatic method for measuring serum mannitol and its use in hemodialysis patients. We evaluated two chemical methods for quantifying mannitol in serum, based on the oxidation of mannitol by periodate, and measurement of the formaldehyde formed with chromotropic acid (colorimetry) or acetylacetone (fluorometry). We found interference in these methods by serum glycerol. Additionally, a high-performance liquid chromatography (HPLC) method was evaluated and found to be specific but impractical for routine use. We therefore, developed an enzymatic fluorometric procedure, based on the oxidation of mannitol by beta-NAD to fructose and NADH, in the presence of the enzyme mannitol dehydrogenase (MD). MD is not commercially available and was partially purified from cultures of Leuconostoc mesenteroides. This new method is specific, sensitive, simple, and accurate and is proposed as the method of choice for measuring mannitol in the serum of patients who received this sugar alcohol during routine hemodialysis treatment.


Clinical significance of serum 7S collagen in various liver diseases. Serum type IV collagen fragment (7S collagen domain) was measured in 30 controls and 152 liver disease patients with a radioimmunoassay using a polyclonal antibody to human placenta 7S collagen. The serum concentrations of 7S collagen (mean +/- SD) were 4.2 +/- 0.9 ng/mL in controls, 5.1 +/- 2.0 ng/mL in acute hepatitis, 6.5 +/- 2.5 ng/mL in chronic inactive hepatitis, 9.5 +/- 3.8 ng/mL in chronic active hepatitis, 14.4 +/- 7.5 ng/mL in liver cirrhosis, and 14.4 +/- 6.9 ng/mL in hepatocellular carcinoma. In acute hepatitis, 7S collagen was slightly increased, whereas type III procollagen N-peptide and prolyl hydroxylase were markedly increased. In chronic liver disease, 7S collagen concentrations increased with the severity of the disease, and also reflected the degree of fibrosis. The serum 7S collagen concentrations were significantly correlated with those of type III procollagen N-peptide and prolyl hydroxylase in all subjects. These results suggest that serum 7S collagen concentration is a useful diagnostic aid for determining hepatic collagen metabolism in liver diseases.


Peritumoral cerebral edema in meningiomas: the role of the tumor-brain interface. We investigated the role of the tumor-brain interface in the production of peritumoral cerebral edema in meningiomas by analysing the size, shape, histological type and location of the tumor, and radiological and operative findings. Our results suggest that changes of the boundary zone between the tumor and the brain such as the disappearance of the subarachnoid space, cortical thinning or loss, and possibly partial loss of the arachnoid membrane are the most important factors in edema production, and that multiple other factors determine the morphological changes which occur at the tumor-brain interface.


Headache and the greater occipital nerve. This paper examines the clinical features of 500 patients with idiopathic headache. Of the 383 patients diagnosed as migraine, it was found that 184 (48%) were suffering from headaches due to irritation of the greater occipital nerve (GON). Such headaches could be arrested by injecting the ipsilateral greater occipital nerve (GON) with local anaesthetic, prevented for up to 4 weeks by injecting 'Depomedrol' into the region of the nerve and for several months by surgical division of the nerve. It is suggested that such patients are not suffering from typical migraine but from headaches due to neural irritation, which, for want of a better name, have been called 'occipital neuralgia'.


Myasthenia gravis developing after discovery of thymoma. We report 4 cases of myasthenia gravis in which myasthenic symptoms developed after a thymoma was known to be present. They include 3 invasive thymomas and one thymoma. In 3 of them, myasthenic symptoms developed 2-7 months after removal of the thymoma; in 1 case, myasthenic symptoms developed 11 years after the thymoma was known to be present. We also discuss here the possible pathogenetic mechanism of post-thymomectomic myasthenia gravis, and propose that it is due to the natural course of the disease.


Increased neurotoxicity of arsenic in methylenetetrahydrofolate reductase deficiency. A 16-year-old girl from Surinam presented with mental deterioration and severe paraparesis with areflexia and bilateral Babinski signs. Laboratory examination showed a hyperhomocysteinemia that was caused by 5,10-methylene-tetrahydrofolate reductase (MTHFR) deficiency. In addition, urine samples contained large amounts of arsenic. An open bag with the pesticide copper acetate arsenite was found to be the source of exposure. In remethylation defects such as MTHFR deficiency, the concentration of methyldonors is severely reduced. As arsenic is detoxified by methylation, we suggest that the MTHFR deficiency in this girl might explain the fact that of all family members exposed to arsenic, only she developed severe clinical signs and symptoms of arsenic poisoning.


Angiotropic lymphoma (intravascular large cell lymphoma) presenting with cauda equina syndrome. A 50-year-old man developed cauda equina syndrome of unknown etiology that was stable for 20 months. Two months prior to sudden death, he experienced new back pain, confusion, seizures, and multiple cranial nerve palsies. Neuropathologic examination revealed angiotropic lymphoma without parenchymal involvement or infarcts in the brain, spinal cord, and muscle. In addition, nerve roots in the cauda equina contained angiotropic lymphoma and infarcts of various ages. Angiotropic lymphoma should be considered as a cause of cauda equina syndrome and of disorders that affect the central and peripheral nervous systems concurrently.


Progressive dementia, without cerebral hemorrhage, in a patient with hereditary cerebral amyloid angiopathy. A now 58-year-old female patient, carrier of the point-mutation in the beta-amyloid gene on chromosome 21 which causes hereditary cerebral hemorrhage with amyloidosis - Dutch type, developed progressive dementia after the age of 55 years. She never suffered from a cerebral hemorrhage. Dementia has been described as a feature of hereditary amyloid angiopathy before, but only in patients who also suffer from strokes. The clinical manifestation of the patient described here underlines the relation between the Dutch type of hereditary amyloid angiopathy and (familial) Alzheimer's disease.


Macro-aneurysm in the basal ganglia region. A 59-year-old female patient suddenly developed vomiting and gait disturbances followed by decreasing consciousness. CT scans revealed a hemorrhage within the left basal ganglia region with rupture into the ventricles and consecutive hydrocephalus. On angiography an aneurysm in the region of the caput nuclei caudati was shown to be the source of the bleeding. On repeat-angiography 4 months later the aneurysm was no longer visualized, probably due to thrombosis. This is an extraordinary case of a basal ganglia aneurysm comparable with the aneurysms of Willis' circle, but located in a region where generally microaneurysms--mostly combined with hypertension or moyamoya disease--can be found.


[Floating intracardiac metastasis].

[Floating intracardiac metastasis]. Two years after the removal of an embryonic testicular tumour from a now 26-year-old man, his general health deteriorated and a holosystolic murmur was heard over the tricuspid valve area. A chest X-ray film demonstrated lung metastases. Biochemical tests were within normal limits, but the human chorionic gonadotrophin level was elevated to 81,030 mU/ml. Echocardiography revealed mild tricuspid regurgitation and a floating mass between the right atrium and ventricle, in contact with the tricuspid valve. The described findings did not support the diagnosis of embolus or inflammatory vegetation. Because of the danger of embolization and obstruction of the pulmonary outflow tract the mass was removed surgically. Enzyme-histological examination identified the tissue as a metastasis of the testicular tumour. The patient's condition at first improved, but he soon died of bleeding from the lung which was infiltrated by trophoblast-containing metastases of the testicular tumour.


An inhibitor of sweet taste response modulates glucose-induced phosphoinositide breakdown in rat pancreatic islets. We studied the effect of a specific-competitive inhibitor of the sucrose taste response, p-nitrophenyl-D-glucopyranoside (PNP-Glu) on insulin release and phosphoinositide metabolism in rat pancreatic islets. The alpha-anomer, but not the beta-anomer, of PNP-Glu at a concentration of 5 mM inhibited insulin release induced by 10 mM glucose. Islets were labeled by exposure for 2 h to 10 uCi of myo-[2-3H] inositol solution supplemented with 2.8 mM glucose. Forty islets were then incubated in the presence of 10 mM LiCl, 1 mM inositol and 10 mM glucose with or without the anomers of PNP-Glu. [3H] radioactivity in the incubation medium remained significantly greater in the presence of the alpha-anomer of PNP-Glu than in the presence of glucose alone after 5- and 20-min incubation. The inositol monophosphate levels in the islets incubated with glucose alone were increased more than in the islets with alpha-anomer. The beta-anomer of PNP-Glu did not change either glucose-induced insulin release or phosphoinositide breakdown. A patch-clamp study revealed that neither anomer affected the glucose-dependent ATP-sensitive K(+)-channels. These results indicate that the anomeric preference for glucose in insulin release in the pancreatic islets is closely associated with phosphoinositide breakdown.


A comparison of commercially available monoclonal antibodies for direct and indirect immunofluorescence culture confirmation and direct detection of parainfluenza viruses. Two commercially available immunofluorescence monoclonal antibody (MAB) reagents (Bartels, Baxter Healthcare, Issaquah, WA; and Symex, Broken Arrow, OK) were evaluated as a means for detecting parainfluenza virus (PIV) both in shell-vial cultures and directly in clinical specimens. Bartels reagents are used in an indirect immunofluorescence assay (IFA) format and exist as MABs reactive with all three PIV serotypes, individually and in a pool. Symex reagents, also available individually and in a trivalent pool, are used in a direct immunofluorescence assay (DFA) format. Among a total of 299 respiratory specimens, 24 yielded PIV. In a shell-vial culture confirmation test, both the individual and pooled monoclonal antibody reagents from both Bartels and Symex detected all 24 PIV isolates. There were three apparent false-positive results with the Bartels pooled IFA reagents. Of the 299 specimens, 160 were also tested directly for the presence of PIV. There were 13 positive specimens among these 160. The Bartels and Symex monoclonal antibody reagents detected similar percentages of positive samples when used for direct detection (that is, 78.6-85.7). No false-positive results were obtained with any of the reagents in the direct-detection format.


A rapid culture alternative to the shell-vial method for the detection of herpes simplex virus. The rapid test for detection of herpes simplex virus (HSV) in clinical specimens based on infection of cells in suspension (SI test) was compared to the shell-vial culture (SVC) method and conventional culture. Mink lung cells were used throughout the study. Detection of HSV was not significantly different whether using SI or SVC. The sensitivity of SI in detecting HSV, when compared with conventional culture, was 93.0% using 0.1 ml inocula and 98.3% using 0.5 ml inocula. The time to obtain a final result with both SI and SVC was 1 day compared with 1-7 days by conventional culture. The SI method detected both HSV type-1 and HSV type-2 clinical isolates. The SI technique is a simple method for the rapid detection of HSV and can yield diagnostic results with a minimum of technical manipulations.


Effects of fluoroquinolone antimicrobials alone and in conjunction with theophylline on seizures in amygdaloid kindled rats. Mechanistic and pharmacokinetic study. The influence of three fluoroquinolone (FQ) antimicrobial drugs (ciprofloxacin (CP), norfloxacin (NF), enrofloxacin (EF)) on seizure parameters in amygdaloid kindled rats was investigated. CP and NF (100 mg/kg i.p.) did not modify seizure parameters while EF induced a decrease in seizure activity. Since clinical data indicate a seizure enhancing interaction between FQ and theophylline (THEO) we studied the influence of concurrent FQ-THEO administration in kindled rats. CP and NF, but not EF given concurrently with a non-seizure modulating dose of THEO (10 mg/kg i.p.) caused increases in seizure activity and aggressiveness in the animals. The CP-THEO induced seizure enhancement was antagonized by 2-chloroadenosine and diazepam. Pharmacokinetic studies demonstrated that THEO serum levels and elimination were not altered by concurrent CP administration. We conclude that coadministration of FQ-THEO can aggravate amygdala kindled seizures and that this aggravation may involve centrally mediated mechanisms.


Laboratory diagnosis of latent human papillomavirus infection. The etiologic association of human papillomavirus (HPV) with uterine cervical cancer has prompted the need for improved laboratory diagnosis of this virus. The application of conventional hybridization technology, including filter in situ hybridization (FISH) and Southern-blot analysis, has revealed that the detection and typing of the virus is inconsistent between sequential specimens from the same individual. To determine whether the polymerase chain reaction (PCR) can be used to provide a more accurate assessment of infection status, two exfoliated cervical cell specimens obtained sequentially from a cohort of 30 women without clinical evidence of cervical abnormalities were analyzed in parallel by FISH and PCR at 6-month intervals. Neither of the procedures provided consistent findings with two sequential specimens suggesting that multiple analyses are necessary to assess infection accurately. However, PCR was less subjective in interpretation and demonstrated greater specificity than did FISH. With the increased sensitivity inherent to PCR, our findings indicated that PCR is more likely to identify latent HPV infection with a single specimen.


Carcinoma in the third testis in a case of polyorchidism and persistent müllerian structure syndrome. Polyorchidism is a rare anomaly and neoplasia arising in a supernumerary testis is even rarer. We herein describe a case of tumor in a supernumerary abdominal testis in an adult male with unrecognized persistent müllerian structure syndrome.


Leukotriene B4 level in stimulated blood neutrophils and alveolar macrophages from healthy and asthmatic subjects. Effect of beta-2 agonist therapy. Leukotriene B4 levels were measured after stimulation by calcium ionophore A23187: (i) in peripheral, neutrophils (PMN) from allergic asthmatics, rhinitis and healthy subjects; (ii) in macrophages collected by bronchoalveolar lavage. LTB4 levels in PMNs were significantly higher in non-treated allergic asthmatics and non-treated subjects with rhinitis compared to controls. Beta-2 agonist-treated asthmatics showed a significantly decreased LTB4 production which was not different from those of controls. In vitro, LTB4 production decreased significantly after PMN incubation with Salbutamol (10(-6) mol l-1). LTB4 produced by AM collected by BAL was measured in non-treated (n = 5) and treated (n = 11) asthmatics with inhaled beta-2 agonist. AM collected from all controls and non-treated asthmatics produced LTB4. By contrast, no production of LTB4 was observed in the treated group. LTB4 production decreased when normal AM were incubated in vitro with Salbutamol (10(-8) mol l-1). These results suggest that biochemical differences occur in PMN and macrophages from subjects treated with beta-2 agonist, presumably in changing the 5-lipoxygenase pathway.


NG-methyl-L-arginine decreases contractility, cGMP and cAMP in isoproterenol-stimulated rat hearts in vitro. NG-Methyl-L-arginine (NMA), an inhibitor of nitric oxide synthesis by vascular endothelium, depresses cardiac function and causes systemic vasoconstriction in vivo. The mechanism of cardiac depression is unclear. Since cGMP inhibits one isoform of myocardial phosphodiesterase (PDE), we hypothesized that a decrease in cGMP might increase PDE activity and lower myocardial cAMP levels, resulting in decreased contractility. Experiments were conducted in isolated, paced, Langendorff-perfused (constant flow) rat hearts under control or isoproterenol-stimulated conditions. In non-stimulated hearts, a 15 min infusion of 30 microM NMA had no effect on cAMP content or on left ventricular dP/dt; however, myocardial cGMP content was decreased. Infusion of 0.01 microM isoproterenol caused dP/dt to increase and caused coronary resistance to fall; myocardial cAMP levels increased while cGMP remained unchanged by isoproterenol. In this stimulated condition, infusion of 30 microM NMA decreased dP/dt and myocardial cGMP and cAMP concentrations. NMA caused coronary resistance to increase to similar maximal values in isoproterenol-stimulated and non-stimulated hearts. Although coronary flow was kept constant during NMA administration, NMA depressed cardiac contractility in isoproterenol-stimulated hearts, but not in non-stimulated hearts, and the depressed contractility in isoproterenol-treated hearts was associated with a decrease in myocardial content of cGMP and cAMP. Therefore, these results are consistent with the hypothesis that NMA may decrease myocardial contractility by decreasing cGMP which leads to increased PDE activity and decreased cAMP.


Nitric oxide donors increase mucus gel thickness in rat stomach. Instillation of the nitric oxide (NO) generator isosorbide dinitrate (0.1-1 mM) into the rat gastric lumen in vivo produced a dose-related increase in mucus gel thickness that was prevented by coadministration of oxyhaemoglobin (10 microM). Isosorbide dinitrate did not induce epithelial cell damage. S-Nitroso-N-acetyl-penicillamine (0.3 mM) and dibutyryl cyclic GMP (1 mM) also increased mucus thickness. These findings, along with the presence of NO synthase in the gastric mucosa, imply a role for NO in vivo in mediation of gastric mucus release.


Influence of temperature on the effects of mu-, delta- and kappa-opioid receptor agonists in the guinea-pig ileum myenteric plexus. Electrically stimulated guinea-pig ileum myenteric plexus-longitudinal muscle was used to determine if changes in temperature alter the inhibitory effects of DAGO ([D-Ala2,N-MePhe4,Gly5-ol]enkephalin, mu-agonist), DPDPE ([D-Pen2,-Pen5] enkephalin, delta-agonist) and U-50,488H (trans-3,4-dichloro)-N-methyl-N-[2-(1- pyrrolidynyl)cyclohexyl]benzeneacetamide methane sulfonate, kappa-agonist). The potency (expressed as the concentration which produces 50% inhibition, IC50) of DAGO and DPDPE was significantly (P < 0.05) decreased at 30 degrees C (8.8 +/- 2.7 x 10(-9) and 8325.2 +/- 1070 x 10(-9) M), when compared to the potency at 37 degrees C (3.8 +/- 0.3 x 10(-9) and 6298.6 +/- 320 x 10(-9) M). Higher temperature (40 degrees C) did not modify the potency of DAGO or DPDPE compared to that at 37 degrees C. However, the potency of U-50,488H was significantly (P < 0.01) increased at 40 degrees C (0.7 +/- 0.0 x 10(-9) M) versus 37 degrees C (2.4 +/- 0.9 x 10(-9) M) or 30 degrees C (2.5 +/- 0.3 x 10(-9) M). The kappa-agonist was more potent than DAGO or DPDPE at 30 or 40 degrees C. These data demonstrate that changes in temperature can alter the potency of opioid agonists.


Reduced contractile responses to forskolin and a cyclic AMP analogue in myocytes from failing human ventricle. Myocytes were isolated from the right or left ventricles of failing and non-failing human hearts. Contractile responses to increasing concentrations of Ca2+, isoprenaline, forskolin and dibutyryl cyclic AMP (a lipophilic analogue of cyclic AMP) were determined. Responses were correlated with the age of the patient, and the severity of failure as defined by New York Heart Association class of symptoms (NYHA), left ventricular ejection fraction (LVEF), left ventricular end diastolic pressure (LVEDP) and dose of diuretics prescribed (diuretic class). The maximum contraction amplitude in high Ca2+ did not change with either age or severity of failure (n = 31-40 patients). Responses to isoprenaline (relative to Ca2+ in the same cell, isoprenaline/calcium ratio) decreased with increasing age (P < 0.001, n = 38), and increasing severity of disease (NYHA, P < 0.001, n = 38; LVEF, P < 0.001, n = 34; LVEDP, P < 0.001, n = 30; diuretic class, P < 0.01, n = 36). The decrease in forskolin/calcium ratio also correlated with age (n = 17, P < 0.005) and increasing severity (NYHA, P < 0.002, n = 17; LVEF, P < 0.05, n = 15; LVEDP, P < 0.02, n = 14; diuretic class, P < 0.05, n = 15). Multiple regression indicated that the contribution of age was greater than that of disease severity for both isoprenaline and forskolin responses. The dibutyryl cyclic AMP/calcium ratio did not change significantly with the age of the patient (P > 0.1, n = 13), or severity as defined by LVEDP (P = 0.05-0.1, n = 12) but did decrease with increasing NYHA class (P < 0.01, n = 13) or diuretics (P < 0.02, n = 12) or with low LVEF (P < 0.002, n = 12). Overall, neither forskolin nor dibutyryl cyclic AMP produced maximum responses greater than isoprenaline in myocytes from failing hearts. Where the response to isoprenaline was not limited by the appearance of arrhythmias, forskolin or dibutyryl cyclic AMP could give a significant (but small) increase in contraction over that with isoprenaline alone. These results provide evidence for a post-receptor defect in addition to beta-adrenoceptor desensitisation in myocytes from failing human heart.


Pulmonary actions of LY255283, a leukotriene B4 receptor antagonist. The actions of LY255283, a leukotriene (LT) B4 receptor antagonist, were examined on guinea pig lung. LTB4 and LY255283 displaced [3H]LTB4 from its binding site on lung membranes with pKi values of 9.9 and 7.0, respectively. In the functional correlate of the binding studies, LY255283 competitively reduced contractile responses of lung parenchyma to LTB4 (pA2 = 7.2). LTB4 produced airway obstruction which was reduced by LY255283 administered i.v. (ED50 = 2.8 mg/kg) or orally (ED50 = 11.0 mg/kg). Contractile responses to histamine, LTD4 and the thromboxane mimetic, U46619, were not reduced by LY255283. The compound also did not inhibit cyclooxygenase or 5-lipoxygenase enzymes. We conclude that LY255283 selectively antagonized pharmacologic responses to LTB4 on lung tissue and appears to be a useful tool to investigate the role of LTB4 in pulmonary disease.


Monoarthritis induces complex changes in mu-, delta- and kappa-opioid binding sites in the superficial dorsal horn of the rat spinal cord. Recently, an experimental model of monoarthritis was described in the rat induced by injection with Freund's adjuvant of the tibio-tarsal joint of one hindlimb. After injection, the clinical and behavioural signs of arthritis are stable from weeks 2 to 6 post-injection. Our purpose was to study the regulation of mu-, delta- and kappa-opioid binding sites in the superficial layers (laminae I-II) of the lumbar and cervical enlargements of the spinal cord 2, 4 and 6 weeks post-injection. Using quantitative receptor autoradiography and highly selective opioid ligands, we found complex changes consisting of a bilateral increase in specific [3H]DAMGO (Tyr*-D-Ala-Gly-NMe-Phe-Gly-ol) and [3H]pCl-DPDPE (Tyr*-D-Pen-Gly-Cl-Phe-D-Pen) binding at 2 weeks post-injection and a bilateral decrease in [3H]U-69593 ((5 alpha,7 alpha,8 beta)-(-)-N-methyl-N-[7-(1-pyrrolidinyl)-1- oxaspiro(4,5)dec-8-yl]) specific binding at 4 weeks post-injection. These changes were restricted to the lumbar level. At 6 weeks post-injection, there was a bilateral increase in [3H]pCl-DPDPE specific binding at both lumbar and cervical levels. Altogether, these results suggest that, after probable local changes in endogenous opioid peptides, the three types of opioid binding sites are differentially involved in the development of the pathological process. These results contrast with the lack of significant modification in mu-, delta- and kappa-opioid binding classically reported at various levels of the spinal cord in polyarthritic rats at 3 weeks post-injection and verified for 2, 4 and 6 weeks post-injection in the present study.


Central noradrenergic inhibition of gastric motility in rats. The roles of central noradrenergic mechanisms in the regulation of gastric motility were investigated in urethane-anesthetized rats in whom an intragastric balloon had been placed. Noradrenaline 100 nmol administered intracerebroventricularly (i.c.v.) significantly decreased spontaneous contractions of the stomach and this decrease in gastric motility was not observed in bilaterally vagotomized animals. Intracisternally administered (i.c.) noradrenaline (10 nmol) induced a decrease in gastric motility comparable to that elicited by noradrenaline 100 nmol i.c.v. Phentolamine 10 nmol i.c. but not the same dose of propranolol i.c. significantly antagonized the noradrenaline (10 nmol)-induced inhibition of gastric motility. These results suggest that alpha-adrenoceptor-mediated noradrenergic mechanisms in the brain stem are involved in the inhibitory regulation of gastric motility.


Neutrophil infiltration into the ischaemic/reperfused rabbit isolated myocardium: effect of PF-5901 and cycloheximide. The Langendorff-perfused rabbit heart preparation has been used to study the interaction of isolated rabbit neutrophils with regionally ischaemic myocardium. Short durations of regional ischaemia (10-60 min) and subsequent reperfusion (30 min) of the hearts with neutrophils resulted in a significant time-dependent accumulation of neutrophils (as assessed by myeloperoxidase activity) in the area at risk. Pre-activation of neutrophils with zymosan-activated serum prior to their infusion into the myocardium potentiated neutrophil accumulation in the area at risk. Pretreatment of the myocardium with a lipoxygenase inhibitor, PF-5901 (10 microM), or a de novo protein synthesis inhibitor, cycloheximide (10 microM), significantly reduced the accumulation of neutrophils in the ischaemic/reperfused myocardium. In contrast, pretreatment of neutrophils with cycloheximide (10 microM, for 15 min) prior to their infusion had no significant effect on neutrophil accumulation in the area at risk. The cyclooxygenase inhibitor, indomethacin (10 microM), had no effect on neutrophil accumulation in the area at risk following ischaemia and reperfusion. These results suggest the involvement of de novo protein synthesis and the lipoxygenase products in the infiltration of neutrophils following ischaemia and reperfusion in vitro.


gamma-Hydroxybutyrate depresses monosynaptic excitatory and inhibitory postsynaptic potentials in rat hippocampal slices. The action of gamma-hydroxybutyrate was studied on pre- and postsynaptic GABA(B) receptors in rat hippocampal neurones in vitro using intracellular recording. gamma-Hydroxybutyrate (1-10 mM) caused a 4-8 mV hyperpolarization of CA1 cells and a 20-80% decrease in monosynaptic excitatory and inhibitory postsynaptic potentials in a concentration-dependent manner. These actions were reversibly inhibited by a novel and selective GABA(B) antagonist, CGP 36742 (20-500 microM) suggesting that gamma-hydroxybutyrate can activate presynaptic as well as postsynaptic GABA(B) receptors.


Impact of polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls on human and environmental health, with special emphasis on application of the toxic equivalency factor concept. A scientific evaluation was made of the mechanisms of action of polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls. Distinction is made between the aryl-hydrocarbon (Ah) receptor-mediated and non-Ah receptor-mediated toxic responses. Special attention is paid to the applicability of the toxic equivalency factor (TEF) concept.


Virus-induced airway hyperresponsiveness in guinea pigs in vivo: study of broncho-alveolar cell number and activity. Recently, we demonstrated that an increased airway responsiveness in vitro can be measured 4, 8, and 16 days, but not 2 days, after intratracheal inoculation of parainfluenza-3 (PI-3) virus to guinea pigs. In the present study airway responsiveness was measured in vivo, and the number, types and activity of broncho-alveolar cells was determined. A significant increase in airflow resistance was measured in spontaneously breathing anesthetized guinea pigs in response to histamine and methacholine, 4 and 8 days after PI-3 virus inoculation. 2 days after inoculation with control solution or PI-3 virus, no difference in the total number of inflammatory cells was observed in the broncho-alveolar lavage fluid. In contrast, on days 4, 8, and 16 after infection a significant increase in the number of alveolar macrophages (102%, 76%, 68%, respectively), monocytes (552%, 374%, 360%, respectively), and lymphocytes (253%, 675%, 396%, respectively) was found. The number of eosinophils was increased as well, but faded with time (378%, 312%, 63%, respectively). PI-3 virus was found to be a very potent activator of broncho-alveolar cells as measured by chemiluminescence. The increase in chemiluminescence production in response to PI-3 virus was reduced in cells obtained from PI-3 virus pretreated animals (day 2, 42%; day 4, 65%; day 8, 22%; and day 16, 30%). In conclusion, PI-3 virus can stimulate broncho-alveolar cells and the virus-induced airway hyperresponsiveness is associated with an influx of inflammatory cells in the respiratory tract.


Evidence for parallel Na(+)-H+ and Na(+)-dependent Cl(-)-HCO3- exchangers in cultured bovine lens cells. BCECF, a cell-entrapable dye with a pH-sensitive fluorescence spectrum, was used to identify transport mechanisms contributing to pH homeostasis of cultured bovine lens epithelial cells. Cells from a spontaneously established lineage were grown on glass coverslips that fit diagonally in a standard curvette and intracellular pH (pHi) was measured. Under perfusion with a CO2-HCO3(-)-free medium (pH 7.45), pHi was 7.19 +/- 0.21 (mean +/- S.D., n = 94 cell preparations). Cell acidifications (pHi to 6.65, n = 8) induced by the 'NH(4+)-loading' method were rapidly followed by a Na(+)-dependent, amiloride-inhibitable pHi recovery. Introduction of a CO2-HCO3(-)-rich medium (pH 7.45) resulted in a small acidification (0.18 +/- 0.04 U, n = 16; P < 0.002) due to rapid CO2 entry and an ensuing slow alkalinization to a pHi near the control CO2-HCO3(-)-free value. Subsequent removal of Cl- resulted in a further alkalinization of 0.18 +/- 0.02 U (n = 13; P < 0.001). This Cl- effect was completely inhibited by the absence of Na+, but was insensitive to amiloride, suggesting the presence of a Na(+)-dependent Cl(-)-HCO3- exchanger. Consistent with this posit, the reintroduction of Na+ to cells perfused in the absence of the cation with a HCO3(-)-containing, amiloride-complemented solution resulted in a gradual recovery from the acidic pHi induced by the baseline conditions (n = 6). The amiloride-insensitive, Na(+)- and HCO3(-)-dependent recovery was completely inhibited in cells pre-incubated with DIDS.(ABSTRACT TRUNCATED AT 250 WORDS)


Enhanced superoxide radical production by stimulated polymorphonuclear leukocytes in a cat model of diabetes. This study examines the possibility that polymorphonuclear leukocyte activation, which can cause endothelial injury, may contribute to the capillary closure of diabetic retinopathy. To examine diabetes-related alterations in polymorphonuclear leukocyte activation, we compared the production of superoxide radical by these cells from normal and from diabetic cats that were maintained hyperglycemic. Polymorphonuclear leukocytes isolated from five diabetic and five normal cats were stimulated with 10 ng ml-1 phorbol myristate acetate, and the maximum rate of their superoxide radical production was measured spectrophotometrically. Stimulated polymorphonuclear leukocytes from diabetic cats generated more superoxide radical, at significantly higher rates, than did those from normals (3.32 +/- 0.33 and 2.50 +/- 0.41 nmol O2- min-1 10(-6) cells, respectively; P < 0.02). While addition of insulin or glucagon did not alter stimulated polymorphonuclear leukocyte radical production, glucose in high concentration did mildly impair its production in both groups. The exaggerated respiratory burst of polymorphonuclear leukocytes in diabetes could contribute to microvascular injury in the retina as well as in other tissues.


Phyllodes tumor of the breast: a multicenter series of 59 cases. Coordinating Center and Writing Committee of FONCAM (National Task Force for Breast Cancer), Italy. A series of 59 phyllodes tumors of the breast was retrospectively reviewed (average follow-up = 3.9 years). Clinical features (age, size of tumor) and diagnostic tests (palpation, mammography, sonography and cytology) were found to be inaccurate in predicting benign (n = 22), borderline (n = 12) or malignant (n = 25) histological type. Limited surgery was associated with a relatively high proportion of local recurrence (enucleation/enucleoresection = 3/5, wide resection = 12/30) compared with mastectomy (2/24). No significant association was observed between the probability of local recurrence and patient's age, histological type or lesion size. Although the study confirms that limited surgery may cure phyllodes tumor, careful follow-up of all patients is needed, since no reliable risk factors for recurrence are available. In malignant cases, axillary node involvement was nil and distant metastases were infrequently observed (3/25). Axillary dissection and search for asymptomatic metastases is not recommended.


Synchronous male breast carcinoma and soft tissue sarcoma occurring within a cancer family. Male breast carcinoma is uncommon and accounts for less than 1% of all cases of breast carcinoma. Soft tissue sarcomas also occur infrequently and cause less than 1% of all cancer related deaths. We report a case of breast carcinoma and soft tissue sarcoma occurring simultaneously in a 34-year-old male with a family history of gastrointestinal neoplasia suggestive of Cancer Family Syndrome. Features, diagnosis and management of this syndrome are reviewed.


An unusual recurrence to the neck after complete resection of hepatocellular carcinoma. A case of hepatocellular carcinoma is presented. Work up at the time of resection revealed no evidence of metastasis. Two months later the patient developed an unresectable mass in his neck which invaded the second cervical vertebral body. This unusual metastasis is an indication that as one site of tumor is increasingly successfully cleared, we are likely to find more examples of tumor spread not seen previously.


[An analysis of the joint activities of primates in a situation of delayed spatial choice]. A leader-dominant baboon of a couple took practically all the food in the situation of spatial delayed choice. The subdominant baboon managed to get some food, too, using fast changes of behavioral tactics during the experimental session. The obvious negative state of the subdominant baboon resulted in a severe neurotic breakdown. Long delays in the experimental scheme enabled the subdominant to get up to 50% of the food reinforcement due to the dominant baboons mistakes. The role of changes in the tactics of the monkeys is discussed.


[The effect of pregnancy on transcallosal evoked potentials]. Pregnancy induced an intrahemispheric redistribution of transcallosal flows mainly in the projection areas of the cat cortex. Functional interhemispheric asymmetry (FIA) of transcallosal evoked responses (TEPs) was determined by interhemispheric distribution of negative-positive responses in non-pregnant cats, whereas in pregnant ones it was determined by the asymmetry of positive-negative TEPs. The right hemisphere played the major part in the changes of the TEPs' amplitude parameters, and the left one--in temporal parameters.


[The effect of hypoxia on the function of the hypothalamic neurosecretory system in rats]. Adaptation to intense hypoxia occurs owing to activation of vasopressin-synthetizing magnocellular neurons of the hypothalamus' paraventricular nucleus. The stability of the organism resistance against hypoxia seems to result from highly active state of the CRF and CRF/vasopressin-synthetizing neurons of the parvocellular portion of the paraventricular nucleus controlling the pituitary-adrenal axis.


[The effect of activation of the superior colliculus on the convulsive activity during picrotoxin-induced kindling]. Bilateral administration of picrotoxin, bicucullin or benzilpenicillin to the colliculi superior induced high-amplitude hypersynchronous discharges in rats. In these conditions, a suppression of generalized seizures induced with picrotoxin, was observed. The superior colliculi seem to play a significant part in antiepileptic system, the part being realized through generator mechanism.


[The comparative characteristics of the background neuronal activity of the central cerebellar nuclei in the alert cat]. Neuronal background activity of the cerebellar central nuclei was studied in alert cats. Statistical analysis revealed some differences in the activity due, probably, to the nuclei's cytoarchitectonic specifics and diversity of afferent cerebellar cortical inputs.


[The possible function of Schwann cells in neuromuscular transmission in the frog]. Anti-galactoside rabbit antisera increased the frequency of slow (atypical) MEPPs. No lysis of the Schwann cells was found in synaptic regions. The data obtained suggest that fixation of antibodies to galactocerebroside on the Schwann cell surface membrane alters the cell's activity thus affecting some characteristics of neuromuscular transmission.


[The role of calcium permeability in shaping the level of the local blood supply to the cerebral cortex]. Cerebral blood flow and the water content in the brain cortex were measured in rabbits with indwelling electrodes. Calcium antagonists increased the CBF in the motor area and decreased it in other regions of the brain.


[The role of the perivascular spaces in shaping the pulse waves of the intracerebral rheoencephalogram]. The intracerebral rheoencephalogram's pulse waves were recorded in rabbits and cats with indwelling electrodes. The waves' amplitude and form were rather diverse and they were inverted in 25% cases: the systolic peak was directed to an increase in the impedance. The dynamics of the waves reflect complex hemoliquorodynamic interrelations in perivascular space.


[The characteristics of the effect of parathyroid hormone on the mechanical activity of the myocardium in rats with a deficiency of Ca and Mg in the drinking water]. Inotropic and lusitropic effects of the parathyroid hormone (PTH) upon the rat heart ventricle myocardium were studied in conditions of calcium and magnesium deficit in drinking water. The control rats revealed two phases of the hormone effect: the positive and negative ones, whereas the experimental rats only revealed the negative inotropic effect. Both the negative and positive inotropic effects were followed by an acceleration of the relaxation.


[The structure of the hemodynamic response and body resistance to hypoxemia]. The highest resistance against hypoxemia was followed by an increased circulation in abdominal organs with simultaneous reduction of blood demand within the area of thoracic aorta in cats. The prevalence of circulation in vitals with no reduction of oxygen demand is characteristic of animals with the low resistance against hypoxemia.


[The osmotic regulation of gastrin secretion by the isolated rat duodenum]. Activating effect of osmotic irritant agent was suppressed by N- and M-cholinoblocking agents, benzohexonium and pyrenzepine in the rat isolated duodenum. The same was true for phentolamine and not for obsidan. The data obtained suggest that the osmotic regulation of the gastrin secretion is performed through the mechanism of a local intramural reflex with participation of N- and M-cholinoreactive and alpha-adrenoreactive agents.


[The effect of stimulation of the globus pallidus and hypothalamus on diuresis in cats]. Stimulation of the pallidum suppressed the diuresis in cats whereas stimulation of lateral hypothalamus increased it. However, stimulation of anterior hypothalamus exerted a suppressing effect on the diuresis.


[The nature of the changes in "central" body temperature in cold exposures in a model of rabbit thermoregulation]. The heat-physical model of the rabbit body with the homeostasis regulation served for the study of "central" temperature behaviour in a thermochamber with the temperature dropping from 22 degrees C to 2 degrees C. The value and character of the "central" temperature changes depended on the initial level of peripheral temperature. In case of a high level of initial heat state, the temperature drops at the start period of the cooling, whereas in case of a low initial level the temperature of the body's "nucleus" increases. The model experiments illustrate the changes of rectal temperatures at the start of the cooling, provided an area of insensitivity is admitted in the work of the rabbit thermoregulation system.


[The enzymatic barrier of the small intestine]. The small intestine's barrier functions are reviewed. The data on mechanical (passive) and active protective systems of the organism against various antigens, toxic substances and proteins, is presented. An important role of these protective systems as an enzyme apparatus of epithelial and postepithelial layers of the small intestine's mucose, is shown.


[Environmental factors and the functional development of the digestive system]. A single action of cold, heat and malnutrition prompts an early transition of the digestive-transport system from juvenile to adult type. Repeated actions of these factors decrease the hydrolytic-transporting function of the digestive tract, the decrease being age-dependent.


[The effect of food and drug herbal extracts on the hydrolysis and transport of sugars in the rat small intestine under different experimental conditions]. Viola tricolor, Frigonella foenum-graecum, Laurus nobilis were shown to reduce the glucose transport. The same was true for extract of tea leaves. Fructose did not affect the velocity of free glucose absorption, slightly reduced the velocity of sucrose hydrolysis and absorption of produced hexoses and water. Participation of active transport of carbohydrates in absorption processes occurring both in vitro and in vivo, is discussed.


[The characteristics of membrane and intracellular hydrolysis and transport in the small intestine during immobilization]. A close interrelationship was demonstrated for the membrane hydrolysis and glucose transport. The immobilisation stress evoked different changes in the regulatory properties of enzymes. The data obtained suggest that stress reduces the stability of the enzyme transport control of the enterocytes' membrane through various regulatory mechanisms.


[The effect of hydrocortisone on pancreatic hydrolase activity in hungry and fed chicks]. Hydrocortisone was shown to prompt the accumulation of pancreatic enzymes taking part in the processing of food protein components in food-deprived chicks. The quality of the enzymes synthetized under hormonal induction does not differ from that in control groups.


[The effect of hydrocortisone on carbohydrase activity along the intestinal villus in chicks]. Administration of hydrocortisone increased the sucrase and amylolytic activities in the brush border of the enterocytes along the intestinal villus and decreased the maltase activity in fed chicken. Hydrocortisone does not affect the level of maltase activity in food-deprived chicks.


[The comparative physiology of membrane digestion in fish]. Significant species differences were found in the activity of some enzymes realising the processes of membrane digestion in elasmobranchs, Chondrostei and teleosts. Similar differences were found in desorption characteristics of the membrane hydrolases suggesting different localisation of the enzymes on the structures of the enterocyte brush border in different species of fish.


[The characteristics of carbohydrate transport in the intestines of fresh-water fishes]. The maximal level of carbohydrate transport was found in the proximal portion of the intestine and in the distal one in the bream and carp. After 48-hr starvation the carbohydrate transport was decreased along the whole length of the intestine. Anoxia and some inhibiting agents changed the intensity of glucose and galactose transport in the fish intestine.


[The adaptational-compensatory reactions of the digestive system during the development of pancreatic atrophy]. Adaptive-compensatory processes have been revealed in the digestive system in pancreatic atrophy. These processes promote: a) the compensation of the pancreatic enzymatic insufficiency and of the cavitary digestion decrease by the alpha-amylase adsorbed at the small intestinal epithelium, b) the increasing of the intestinal pH, c) the keeping up of the postprandial levels of gastrointestinal hormones.


[The adsorption of pancreatic enzymes by the small intestine as one of the mechanisms coupling cavitary and parietal intestinal digestion]. Intraduodenal administration of inactivated intestinal mucus and erythrocytes abolished the inhibiting effect of pancreatine on the pancreatic secretion in dogs. The enzymes fixed by the duodenum mucose seem to inhibit the pancreatic secretion.


[Changes in the enzyme activities of the pancreas and small intestine in rat pups during the transition from milk to definitive nutrition]. The diet with high contents of protein, lipids and carbohydrates resulted in an adaptive shift of the hydrolytic function of the rat pancreas and small intestine. High-protein diet led to activation of proteolytic enzyme pool, the same occurred with high-lipid and high-carbohydrate diets.


[The dynamic functional-morphological changes in the small intestine of obese patients undergoing weight-reducing diet therapy]. Two groups of pathogenetic mucosal changes peculiar to fasting period were revealed in 28 obese patients: a structural rearrangement of mucosa with inhibition of mitotic activity of epithelial crypts, and the inflammatory responses manifested by increased cellular infiltration of stroma and compensatory-adaptive changes of capillary bed structures. The adequate dietotherapy provided full recovery of the mucosa tissue parameters on the 26th day.


[The detailed characteristics of the enzyme spectrum of the small intestine in rats in the early postnatal period]. The activity of the small intestine's peptide hydrolases is higher in 1-day old rats than in adult rats, whereas levels of activity of alkaline phosphatase and diglycyl glycine peptidase do not differ significantly in these two groups of the rats. Our own data on carbohydrases corroborate other authors' evidence and reveals that activities of lactase, sucrase and maltase are either absent or very low in the first days of life and sharply increase by the third week of postnatal development. Adaptive changes of regulatory properties of lactase and alkaline phosphatase are revealed.


[The enzyme-transport characteristics of the small intestine of rats during aging]. Ageing was shown to be accompanied by changes of the activity levels of the small intestine's carbohydrases, dipeptidases, and alkaline phosphatases. The changes occurred both under normal conditions and after operations on the small intestine. In ageing, the small intestine's capacity for the homeomorphosis decreases irrespective of either increase or decrease in the functional loading. The age involution of the gastrointestinal tract is followed by disturbances not only in hydrolytic processes but in transport those as well: particularly deceleration of glucose accumulation in preparations of the rat intestine mucose. Regulatory properties of some enzymes were changed, too, in old rats.


[Trans-sorption as an important mechanism of molecular transport in biological systems]. The mechanism of trans-sorption was shown to be based on mobile adsorption of various substances at specific and unspecific sites localised in spatially organised systems integrated within premembrane structures. Some specifics and possible physiological significance of trans-sorption are discussed using mathematical models of nutrients transfer across autonomous premembrane layer in the small intestine.


[The transport system that pumps monosaccharides from enterocytes across the basolateral membrane]. Using a new, in vitro, experimental approach the transport of glucose, galactose, fructose and glycine in seroso-mucosal direction in the small intestine, was studied under different modes of oxygenation. In case of monosaccharides under the oxygenation from serosal surface, the substrate concentration is lower in the small intestine tissue than under anoxia. The effect of the oxygenation from serosal surface on the galactose transport was shown to be sensitive to absence of sodium ions, ouabain, phloretin, and not sensitive to phlorizin. The data obtained corroborate the hypothesis of the existence of a mechanism pumping off monosaccharides enterocytes through the basolateral membrane.


[The characteristics of the absorption in the small intestine of 2 casein hydrolysates and of an equivalent amino acid mixture]. Using casein hydrolysates and the amino-acid mixture applied to mucosal or serosal surfaces, their effect on the transport of free glycine, glycine formed during glycyl glycine hydrolysis, and galactose, was studied in the small intestine. The data obtained suggest a mechanism of peptide transport in the enterocytes as well as a possibility of a regulatory effect of the peptides contained in casein hydrolysates on the transport of some basic nutrients.


[The final stages in the hydrolysis of foodstuffs in the small intestine of mammals with different food specializations]. The activity of the small intestine's enzymes realising final stages of nutrients hydrolysis was studied in herbivorous, omnivorous and carnivorous animals. The small intestine's enzyme spectrum corresponded to the food specialisation of the animals. The proximal-distal gradient of the distribution of digestive enzymes along the small intestine has both some similar features and certain differences.


[The regulation of specialized appetites]. Motivational-emotiogenic systems of the brain were shown to take part in regulation of specific appetite. The taste seems to be not only the control means determining the acceptability of food but a link of the motivational-emotional system as well. The taste aversions seem to play a permanent role in organisation of the feeding behaviour.


[A multichannel transporter for glucose and amino acid transport]. The recording of the short-circuit current responses to glucose or glycine was used to monitor active nutrient transport in the rat small intestine from mucose to serose. The coupled cotransporter for Na+ glucose and glycine was shown to consist of the central ion channel and a few nutrient channels around it. It seems that the combination of some types of ion channels with various types of nutrient channels leads to a formation of multi-channel transporters with different ion-nutrient specifics.


Oncogene transduction and activation.

Oncogene transduction and activation. This article explains what a transducing virus is, summarizes what is currently known about the origins of transducing viruses, considers the frequency of the generation of transducing viruses, and discusses how transduced oncogenes undergo activation. Most of the data come from work with avian leukosis viruses, as research with this group of viruses has set many of the precedents for work on retroviral transduction.


Detection of murine leukemia viruses--in vitro infectivity tests. The four biologically distinct classes of murine leukemia viruses (MuLVs) are described and the potential for expression of each as infectious virus in laboratory mice is discussed. Commonly used in vitro methods for MuLV detection and identification are outlined.


HHV-6 and HHV-7 as exogenous agents in human lymphocytes. Human herpesviruses 6 and 7 (HHV-6 and HHV-7), the newly recognized lymphotropic members of the herpesvirus family, were isolated from peripheral blood mononuclear cells (PBMC) of healthy individuals. HHV-7 was recovered from PBMC after exposing the cells to conditions leading to T cell activation. No virus could be recovered from quiescent non-activated cultures, suggesting that the virus resided latently in these cells and that it could be induced from latency by T cell activation. HHV-6 could not be recovered from PBMC using similar approaches. However, it could be repeatedly reactivated from PBMC after infection of the cells with HHV-7. Upon further passaging in culture, HHV-6 took over the population and virus stocks appeared to be clear of the original HHV-7 helper virus. The results of this study suggest that genomes of the lymphotropic herpesviruses are resident in PBMC of healthy adults. HHV-7 can supply the functions required for its reactivation from latency. It can also act as a helper virus for the reactivation of HHV-6 from latency. Following the initial reactivation step, HHV-6 can replicate autonomously in the cells. These results may be relevant to the findings that HHV-6 infection is induced in immunosuppressed patients and in patients undergoing organ transplantation. Furthermore, these studies may exemplify situations whereby propagation of known agent(s) in vitro results in the activation of human pathogens resident latently in the cells.


Latent parvoviral infection of continuous cell lines. The parvoviruses are a family of single-stranded DNA-containing viruses which are known to establish inapparent infections of continuous, and in some cases, primary cell cultures. Their small size and great stability suggest that they would be difficult to eliminate from a biological component purified from a contaminated cell line. Thus, precautions should be taken to exclude such agents from initial cell cultures, and from the reagents used to maintain them.


Neurotrophic interactions in the nervous system. Traditional views have suggested that information flow in the nervous system is dependent on millisecond-to-millisecond communication occurring in a point-to-point manner. However, recent evidence suggests that growth and trophic functions are central to development and maintenance of function in the brain. In turn, trophism occurs over days to weeks to months and appears to underlie processes as diverse as learning, memory and development, on the one hand, and the pathogenesis of disease, on the other. This work group focussed on molecular and cellular mechanisms underlying trophic function in the brain.


Structural biology and diabetes mellitus: molecular pathogenesis and rational drug design. Emerging concepts in the aetiology and pathogenesis of Type 1 (insulin-dependent) diabetes mellitus may offer new opportunities for treatment and cure. Here we describe recent advances in structural molecular biology and molecular design relevant to rational drug discovery. Such approaches focus on the three-dimensional structures of macromolecules and their interactions. In the coming decade such techniques may be applied to a wide variety of diabetes-related targets.


Investigation of ACTH responses of chickens with autoimmune disease. An altered immunoendocrine feedback regulation within the hypothalamo-pituitary-adrenal axis may modulate the pathogenesis of an avian autoimmune disease. To date studies have been hampered by a lack of reliable, specific, and sensitive methods for determining adrenocorticotropic hormone (ACTH) in chickens. The present study describes the determination of ACTH in plasma of chickens with a commercial radioimmunoassay, the antibody of which binds to the midregion of human ACTH 1-39. The chickens, kept on a 12-hr day and 12-hr night shift with artificial light, showed changes in plasma ACTH concentrations during the light phase with maximum values 8 hr after the light was turned on. ACTH was not measurable after treatment with dexamethasone. Intravenous administration of supernatants from concanavalin A-stimulated spleen cells increased basal plasma ACTH concentrations more than 20-fold within 1 hr. This increase in plasma ACTH was higher and longer lasting in UCD 200 chickens, an animal model for scleroderma, compared with outbred and inbred normal White Leghorn chickens.


Regulation of hepatic growth hormone receptors in coho salmon (Oncorhynchus kisutch). Factors potentially regulating hepatic growth hormone (GH) receptors in coho salmon (Oncorhynchus kisutch) have been investigated. From December to June of the first year, relative changes in hepatic 125I-sGH binding and 35SO4 incorporation by ceratobranchial cartilage were similar. Stunted salmon, which in seawater have elevated plasma GH yet fail to grow, showed lower hepatic 125I-sGH binding than did normally growing seawater salmon. However, MgCl2 treatment of stunts' membranes to reveal total specific binding of 125I-sGH indicated receptor occupation by endogenous sGH. Total specific 125I-sGH binding was low in seawater stunts and remained low if these fish remained unfed after return to fresh water, but increased approximately twofold upon feeding. Total specific binding in fasted salmon in fresh water showed a trend toward decreased levels by 1 week; by 3 weeks, binding was 40% lower than in fed fish. There was a positive correlation (r = 0.600) between condition factor and total specific binding in fed and fasted salmon in fresh water. Two weeks after hypophysectomy total specific binding was 50% lower than in sham-operated control salmon, indicating pituitary regulation of GH receptors. GH treatment reduced both free and total 125I-sGH binding in salmon examined 24 hr after treatment. Treatment with recombinant bovine insulin-like growth factor I, thyroxine, or cortisol did not affect free 125I-sGH binding. Both the pituitary and nutrition appear to be prime regulators of hepatic GH receptors in coho salmon.


VIP inhibits CCK-induced gallbladder contraction involving a beta-adrenoceptor mediated pathway in the rainbow trout, Oncorhynchus mykiss, in vivo. The effects of VIP on CCK-induced contractions of the gallbladder were studied in the rainbow trout, Oncorhynchus mykiss, in vivo. Gallbladder pressures were measured via a catheter inserted into the gallbladder, and drugs were injected through a catheter in the dorsal aorta. CCK-induced contractions of the gallbladder wall were reduced in a dose-dependent manner by VIP. The effect of VIP was inhibited by the beta-adrenergic antagonist sotalol, indicating an adrenergic link in the VIP inhibition of the CCK-induced contraction. It is suggested that activation of VIP nerves could facilitate postprandial filling/refilling of the gallbladder despite the increased plasma levels of CCK during this period.


Molecular genetic studies of a 10.9-kb operon in Escherichia coli for phosphonate uptake and biodegradation. Bacteria that use phosphonates as a phosphorus source must be able to break the stable carbon-phosphorus bond. In Escherichia coli phosphonates are broken down by a C-P lyase that has a broad substrate specificity. Evidence for a lyase is based on in vivo studies of product formation because it has been proven difficult to detect the activity in vitro. By using molecular genetic techniques, we have studied the genes for phosphonate uptake and degradation in E. coli, which are organized in an operon of 14 genes, named phnC to phnP. As expected for genes involved in P acquisition, the phnC-phnP operon is a member of the PHO regulon and is induced many hundred-fold during phosphate limitation. Three gene products (PhnC, PhnD and PhnE) comprise a binding protein-dependent phosphonate transporter, which also transports phosphate, phosphite, and certain phosphate esters such as phosphoserine; two gene products (PhnF and PhnO) may have a role in gene regulation; and nine gene products (PhnG, PhnH, PhnI, PhnJ, PhnK, PhnL, PhnM, PhnN, and PhnP) probably comprise a membrane-associated C-P lyase enzyme complex. Although E. coli can degrade many different phosphonates, the ability to use certain phosphonates appears to be limited by the specificity of the PhnCDE transporter and not by the specificity of the C-P lyase.


Possible function of tRNA(Thr)ACG in regulation of solvent formation in Clostridium acetobutylicum. The mutation of Clostridium acetobutylicum mutant AA2, defective in the formation of acetone and butanol, was shown to be caused by a single insertion of Tn916 close to the structural gene thrA, encoding the tRNA(Thr)ACG. The DNA region containing the thrA gene was cloned and sequenced. Start and end points of the transcript were determined by primer extension and S1-mapping analysis. The results obtained were identical to predictions derived from the DNA sequence by various RNA-analysing computer programs. The rarely used ACG codon seems to be confined to genes expressed at the end of the exponential growth phase or involved in uptake or turnover of minor C or N substrates. Evolutionary aspects of this codon selection and a possible translational regulation mechanism are discussed.


The mechanism of action of chlorhexidine. Chlorhexidine did not inhibit ATPase in intact cells of Escherichia coli K12 W1317i-, even at bactericidal concentrations, and ATP hydrolysis was greatest at the highest concentration (40 mg/l), even though no net uptake of substrate occurred. Like dinitrophenol and tribrominated salicylanilide, polymyxin and chlorhexidine collapsed the membrane potential at inhibitory concentrations. Membrane disruption, and not ATPase inactivation, is considered the lethal event in chlorhexidine action.


Utilisation of glycerol and glycerol 3-phosphate is differently affected by the phosphotransferase system in Bacillus subtilis. Glycerol and glycerol 3-phosphate uptake in Bacillus subtilis does not involve the phosphotransferase system. In spite of this, B. subtilis mutants defective in the general components of the phosphotransferase system, EnzymeI or Hpr, are unable to grow with glycerol as sole carbon and energy source. Here we show that a Hpr mutant can grow on glycerol 3-phosphate and that glycerol 3-phosphate, but not glycerol, can induce glpD encoding glycerol-3-phosphate dehydrogenase. Induction of glpD also requires the glpP gene product which is a regulator of all known glp genes. Thus the phosphotransferase system general components do not interfere with the overall regulation of the glp regulon. Revertants of a Hpr mutant which can grown on glycerol carry mutations closely linked to the glp region at 75 degrees on the B. subtilis chromosomal map. This region contains the glpP, the glpFK and the glpD operons. The glpFK operon encodes the glycerol uptake facilitator (glpF) and glycerol kinase (glpK). The present results demonstrate that one of these genes, or their gene products, is the target for phosphotransferase system control of glycerol utilisation. Furthermore we conclude that utilisation of glycerol and glycerol 3-phosphate is differently affected by the phosphotransferase system in B. subtilis.


Energy conservation in fermentative glutarate degradation by the bacterial strain WoG13. Dicarboxylic acids with 2-5 carbon atoms can be degraded fermentatively by pure cultures of various strictly anaerobic bacteria. The small amount of free energy released in these decarboxylations (about 20-25 kJ mol-1) is conserved as sole source of growth energy either through sodium-pumping decarboxylases or through electrogenic substrate/product transport devices. In the glutarate-fermenting bacterial strain WoG13 a glutaconyl-CoA-decarboxylating enzyme activity was detected. This enzyme was inhibited by avidin and was stimulated by sodium ions. The enzyme activity was partially associated with the cytoplasmic membrane, indicating that energy conservation is accomplished through a sodium-ion-pumping glutaconyl-CoA decarboxylase enzyme.


Inhibitors of lipopolysaccharide biosynthesis impair the virulence potential of Escherichia coli. Inhibition of 3-deoxy-manno-octulosonate cytidylytransferase (CMP-KDO transferase; EC 2.7.7.38) by 8-amino-2,6-anhydro-3,8-dideoxy-D-glycero-D-talo-octonic acid (NH2dKDO) halts the growth of Gram-negative bacteria by depriving the cells of the 3-deoxy-D-manno-2-octulosonate required for the biosynthesis of the core region of the lipopolysaccharide components of the outer membrane. Low levels of this inhibitor increase the vulnerability of Escherichia coli to hydrophobic antibiotics, detergents, the complement-mediated antibacterial activity of serum, phagocytosis, and enhance the rate at which bacteria are cleared from the mouse bloodstream.


Epstein-Barr virus-transformed B lymphocytes produce low molecular mass molecules with autocrine growth factor and competence factor activity. A human Epstein-Barr virus (EBV)-positive lymphoblastoid B cell line, named BA-D10-4, produces a factor of a molecular mass less than 10 kDa that promotes cell proliferation of both BA-D10-4 cells and other human T or B lymphoid cell lines, either EBV-positive or -negative. The factor synergizes with higher molecular mass autocrine growth factors and makes both BA-D10-4 cells and B cell lines from Burkitt's lymphoma, but not cells from T cell leukemia, more responsive to interleukin-1 and interleukin-6. Therefore, this low molecular mass factor seems to be an autocrine growth factor per se and to have the characteristics of a competence factor.


Why do human hepatitis viruses replicate so poorly in cell cultures? The five viruses which classically cause hepatitis in man represent diverse families of viruses and share in common only a striking hepatotropism and substantial restrictions to replication in conventional cell cultures. Hepatitis A virus is unique among these viruses in that it is amenable to propagation in cell culture, but replication of this virus is much slower and less efficient than replication of other picornaviruses. This probably reflects less efficient cap-independent viral translation, as well as restrictions at other points in the replication cycle. We speculate that the significantly restricted replication of hepatitis viruses in cell culture reflects evolutionary forces controlling their transmission and propagation through human populations.


Molecular biology of Bacillus subtilis cytochromes. Bacillus subtilis cells must have cytochromes for growth and can synthesize cytochromes of a-, b-, c-, d-, and o-types. After a long lag, our knowledge of the structure, genetics and specific role for these cytochromes is now growing exponentially as the result of recent research. This progress is reviewed here and includes, for example, the discovery of two different cytochrome a systems and genes required for their biogenesis.


Individual and combined effects of a thromboxane receptor antagonist and different antithrombotic agents in a rat microcirculatory thrombosis model. The antithrombotic effect of a thromboxane A2 receptor antagonist (HN-11501:5-[2-(4-chlorophenylsulfonylamino)-ethyl]-2-thienylox y-acetic acid) alone and in combination with other antithrombotic agents has been studied in an experimental thrombosis model in which laser lesions are used to induce a defined thrombosis in rat mesenteric venules. The thromboxane receptor antagonist showed a significant and dose-dependent antithrombotic effect if given orally. The strongest additive thrombosis-inhibiting effect was observed after oral administration of HN-11501 at a dose of 2.5 mg/kg together with an intravenous infusion of 1 microgram/kg/h of a prostacyclin analogue (cicaprost). An additive antithrombotic effect was also observed after oral application of 2.5 mg/kg of HN-11501 and intravenous injection of 0.2 mg/kg of a low molecular weight heparin (Fraxiparine). The combination of 2.5 mg/kg of HN-11501 orally with an intravenous injection of 0.1 mg/kg molsidomine also had a significant additive effect. No significant additive effect was observed when 2.5 mg/kg of HN-11501 and 10 mg/kg of acetylsalicylic acid were orally administered simultaneously.


[Infections with herpesvirus 6--really only "exanthema subitum"? Part 1: More frequent disease pictures]. The human herpes virus 6 (HHV6) is the causal agent of exanthem subitum (rose rash of infants). In addition, however, a wide range of other clinical manifestations are possible, the more common of these include: Untypical exanthem or uncharacteristic rash (with and without fever), acute respiratory disease (ARD) acute gastroenteritis and febrile convulsions (with and without exanthem). More uncommon, and in part unknown, clinical manifestations associated with HHV6 infection are discussed in Part 2 of this paper. In common with other herpes viruses, HHV6 also tends to show persistence and intermittent or chronic shedding in the normal population, making the unusually early infection of children (seroconversion in the first year of life in up to 80% of all children) understandable. This means that HHV6 infections manifesting in close temporal association with recommended vaccinations may be misinterpreted as complications of the vaccination. Today, the situation can be clarified by employing special virological-serological laboratory tests, which are available throughout the country.


[Infections with herpesvirus 6--really only "exanthema subitum"? Part 2: Rare or unknown disease pictures]. The human herpes virus 6 (HHV 6) may induce not only the wellknown condition of exanthem subitum, but also a number of more common (cf. Part 1) or rare, even previously unknown, clinical manifestations. Part 2 of this paper deals with the more rarely observed manifestations. These include complications of ARD (sinusitis, otitis media, bronchial pneumonia) hepatitis, encephalitis or Pfeiffer's disease (mononucleosis-like syndrome). In individuals with a relevant disposition (genetic HLA/DR type?) initiation or (re-)activation of rheumatoid arthritis (JCA = juvenile chronic arthritis) or chronic iridocyclitis may occur. Although, on account of the high prevalence of vaccination in our population (approximately 95%), prenatal infections are extremely rare, they may manifest in a severe "septic" form (fatalities have occurred) or may lead to neurological deficits (comparable with cytomegalovirus infection). To date, no specific therapy (e.g. gammaglobulin, virostatics) or reliable preventive measures (e.g. vaccination) are available.


Effect of the GABAA agonist muscimol on prolactin secretion from human prolactin-secreting adenomas and GH3 rat pituitary tumour cells. The effect of muscimol, a specific potent GABAA receptor agonist, on prolactin release from human prolactin-secreting tissue was investigated using a perifusion system. Perifusion studies on normal rat anterior pituitary tissue, which has identical GABA receptors to those found in normal human pituitary glands, show that muscimol has a specific biphasic effect on prolactin release. This is characterized by an initial transient stimulation (222.3 +/- 21.6% of basal) lasting for 5-10 min followed by a more prolonged inhibitory phase (63.9 +/- 3.1% inhibition of basal). Five human prolactin-secreting adenomas were studied, and in none of the tumours could a biphasic response be demonstrated. One of the prolactin-secreting adenomas had a blunted inhibitory response, but the other 4 showed no inhibitory effect of muscimol on prolactin release. Muscimol had no significant effect on basal or thyrotropin-releasing-hormone (TRH)-stimulated prolactin secretion from GH3 rat pituitary tumour cells. These studies suggest that the GABAergic effect on prolactin secretion is absent or altered in both rat and human prolactin-secreting tumour cells.


Mu-, delta-, and kappa-opioid receptor agonists selectively modulate sexual behaviors in the female rat: differential dependence on progesterone. Previous studies suggested that opioid receptor agonists infused into the lateral ventricles can inhibit (through mu receptors) or facilitate (through delta receptors) the lordosis behavior of ovariectomized (OVX) rats treated with estrogen and a low dose of progesterone. The present study investigated the behavioral and hormonal specificity of those effects using more selective opioid receptor agonists. Sexually experienced OVX rats were implanted stereotaxically with guide cannulae aimed at the right lateral ventricle. One group of rats was treated with estradiol benzoate (EB, 10 micrograms) 48 hr and progesterone (P, 250 micrograms) 4 hr before testing, whereas the other group was treated with EB alone. Rats were infused with different doses of the selective mu-receptor agonist DAMGO, the selective delta-receptor agonist DPDPE, or the selective kappa-receptor agonist U50-488. The females were placed with a sexually vigorous male in a bilevel chamber (Mendelson and Gorzalka, 1987) for three tests of sexual behavior, beginning 15, 30, and 60 min after each infusion. DAMGO reduced lordosis quotients and magnitudes significantly in rats treated with EB and P, but not in rats treated with EB alone. In contrast, DPDPE and U50-488H increased lordosis quotients and magnitudes significantly in both steroid-treatment groups. Surprisingly, measures of proceptivity, rejection responses, and level changes were not affected significantly by mu or kappa agonists, although proceptivity and rejection responses were affected by DPDPE treatment. These results suggest that the effects of lateral ventricular infusions of opioid receptor agonists on the sexual behavior of female rats are relatively specific to lordosis behavior. Moreover, the facilitation of lordosis behavior by delta- or kappa-receptor agonists is independent of progesterone treatment, whereas the inhibitory effect of mu-receptor agonists on lordosis behavior may require the presence of progesterone.


The gene for the alpha 1 subunit of the skeletal muscle dihydropyridine-sensitive calcium channel (Cchl1a3) maps to mouse chromosome 1. Cchl1a3 encodes the dihydropyridine-sensitive calcium channel alpha 1 subunit isoform predominantly expressed in skeletal muscle. mdg (muscular dysgenesis) has previously been implicated as a mutant allele of this gene. Hybridization of a rat brain cDNA probe for Cchl1a3 to Southern blots of DNAs from a panel of Chinese hamster x mouse somatic cell hybrids suggested that this gene maps to mouse Chromosome 1. Analysis of the progeny of an inbred strain cross-positioned Cchl1a3 1.3 cM proximal to the Pep-3 locus on Chr 1.


Mapping of a human brain voltage-gated calcium channel to human chromosome 12p13-pter. Degenerate DNA oligomers coding for highly conserved regions of the voltage-gated calcium channel were synthesized for the polymerase chain reaction (PCR) using DNA from a human brain cDNA library as template. PCR amplified a 640-bp DNA fragment from the human brain cDNA library. Sequencing revealed that this fragment encodes part of a protein highly homologous to a subtype of the dihydropyridine-sensitive calcium channel cloned from rabbit heart and rat brain. Southern analysis of panels of somatic cell hybrids mapped the 640-bp fragment, CACNL1A1, to human chromosome 12p13-pter.


Expression of cytokines and their receptors by human thymocytes and thymic stromal cells. The repertoire of cytokine and cytokine receptor mRNA expressed by unstimulated human thymocytes and thymic stromal cells was explored by a quantitative polymerase chain reaction (PCR) using sequence specific internal standards. Of the 18 cytokines tested we found a considerable overlap in the expression of cytokines by human thymocytes and by thymic stromal cells; both cell types express the mRNA for interleukin-1 beta(IL-1, IL-6, IL-7 and tumour necrosis factor-alpha (TNF-alpha). However, there are substantial differences in the levels of cytokine mRNA expressed in these two types of cells as revealed by the quantitative PCR assay. Stromal cells express considerably higher levels of IL-1 beta and IL-6 than thymocytes (14- and 27-fold respectively). In addition, a number of cytokines such as lymphotoxin and interferon-gamma (IFN-gamma), are expressed exclusively in thymocytes whereas others such as stem cell factor (SCF), IL-1 receptor antagonist-2 (IRAP-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are produced only in stromal cells. There is a complete overlap in the expression of a group of cytokine receptors tested in thymocytes and thymic stromal cells; these include IL-1R, IL-2R, IL-6R, IL-7R, TNFR and stem cell growth factor receptor (c-KIT). The expression of specific cytokines by thymic stromal cells and the parallel expression of their receptors on thymocytes under physiological conditions, support the hypothesis that these cytokines participate in paracrine interactions between these two cell populations during thymocyte differentiation.


Modulation of leukotriene generation by invasive bacteria. The effect of invasive bacteria on the release of proinflammatory mediators (oxygen radicals, leukotriene release) from human polymorphonuclear neutrophils was studied. Bacterial stimuli were used including genetically cloned invasive Yersinia enterocolitica strains 108-P (bearing the phagocytosis-resistance plasmid) and 108-C (plasmidless variant), Listeria monocytogenes [SLCC 5779 (inv-) and NCTC 7973 (inv+)] as well as an Escherichia coli K 12 strain (pRI 203) in which the inv gene of Y. pseudotuberculosis was cloned. When human polymorphonuclear granulocytes were studied as target cells the inv+ as well as the inv- strains were phagocytosed to a comparable amount with the exception of the L. monocytogenes strain (inv+). Among the invasive strains E. coli HB 101 (pRI 203) was the most active to trigger polymorphonuclear leucocytes (PMN) for oxygen radical production. Preincubation of the cells with bacteria and subsequent stimulation with the Ca ionophore A23187 or opsonized zymosan suppressed the chemiluminescence response to a different degree. The various bacterial strains did not induce leukotriene release from endogenous arachidonic acid. Subsequent stimulation of the infected cells with Ca ionophore or opsonized zymosan led to an altered pattern of the combined amounts of leukotriene B4 (LTB4), 20-OH- and 20-COOH-LTB4 as well as the ratio of LTB4 versus 20-OH and 20-COOH-LTB4. Infection of the cells also reduced strain dependently the number of LTB4-receptor sites. Our data suggest that bacterial uptake modulates the inflammatory response of granulocytes (e.g. chemiluminescence response, leukotriene generation).


Leukotriene A4 modulates generation of leukotriene B4 and sulphidopeptide leukotrienes by human neutrophils. We investigated the influence of exogenous leukotriene A4 (LTA4) on the reactivity of polymorphonuclear leucocytes (PMN). PMN were either prestimulated with LTA4 or incubated simultaneously with LTA4 and the Ca ionophore A23187 or sodium fluoride (NaF). The Ca ionophore A23187 and NaF induced generation of LTB4 from PMN was significantly diminished in the presence of LTA4 while the formation of LTC4 was enhanced. In contrast, preincubation of cells with LTA4 followed by subsequent stimulation with NaF synergistically increased the LTB4 generation from PMN. LTA4, either alone or in combination with the calcium ionophore A23187 or NaF, decreases GTPase activity in human PMN. This decrease was abolished when LTA4 pretreated cells were subsequently stimulated with NaF, but not with calcium ionophore A23187, suggesting a regulatory role of LTA4 on G-proteins. The results demonstrate dual functions of LTA4: it serves as a substrate for the generation of leukotrienes and also regulates the susceptibility of human PMN for subsequent response.


Regulation of production of adrenocorticotropin-like proteins in human mononuclear cells. Cells of the immune system produce biologically active adrenocorticotropic hormone (ACTH). Many laboratories, however, have been unable to replicate experiments which demonstrate ACTH in immune cells. Sensitive immunohistochemical staining and digital scanning, confocal microscopy were used to study regulation of ACTH-like immunoreactivity (ACTH-IR) in human mononuclear cells. Cytoplasmic ACTH-IR was induced by corticotrophin releasing factor (CRF)/arginine vasopressin (AVP), and also by protein kinase C (PKC) activation and by the interferon (IFN-alpha beta inducer, Na-polyinosinic-polycytidylic acid (polyIC). Induction of cytoplasmic ACTH-IR was maximal within 6 hr of stimulation with CRF/AVP or phorbol myristate acetate (PMA). Recombinant human interleukin-1 beta (rhIL-1 beta) was also stimulatory, but rhIL-1 alpha had minimal effect. Regulation of ACTH-IR production in immune cells parallels the regulation of ACTH in the anterior pituitary, and ACTH-like material may affect immune responses.


T lymphocyte responses of sheep to bovine leukaemia virus infection. Sheep were experimentally infected with bovine leukaemia virus (BLV) by inoculation of peripheral blood lymphocytes (PBL) from BLV infected sheep. Monoclonal antibodies were used to monitor changes in lymphocyte subpopulations in the first few weeks after inoculation. The polymerase chain reaction (PCR) detected BLV DNA in PBL of infected sheep 11-15 days after inoculation, that is, before antibodies to viral structural proteins were detected at 15-39 days post-inoculation. A rise in the number of both B and T lymphocytes coincided with detection of infection by PCR. At this time, an increase in the number of circulation CD8+ lymphocytes resulted in a low CD4: CD8 ratio. It appears that in BLV infection there is a host specific cell-mediated immune response to infected lymphocytes rather than a general immune response to foreign antigens. This response, which is characterized by an increase in the number of circulating CD8+ lymphocytes, precedes seroconversion. There is considerable variation between animals in this cytotoxic T lymphocyte response.


T-lymphocyte mediated injury of beta cells in dual-aetiology diabetes mellitus in Swiss albino mouse. Earlier we had described a dual aetiology diabetes mellitus (DADM) in mice injected with a sub-diabetogenic dose of streptozotocin (SD-SZN) and afterwards infected with coxsackie B3 virus (CBV). Further experiments were conducted to understand the mechanism of diabetogenesis. In in vitro stimulation and proliferation tests, the splenic lymphocytes (SLC) of mice given either SD-SZN or CBV infection showed lower responses to two T cell mitogens than those of control mice, indicating an immunosuppressive effect. Unexpectedly, SLC of mice given both SD-SZN and CBV showed enhanced response, indicating immunoactivation; they were not stimulated to proliferation in response to CBV antigen, indicating that the immunoactivation was not directed against CBV, but against streptozotocin or cellular elements. When mice were depleted of T cells by injecting with anti-thymocyte serum, the diabetogenic effect of SD-SZN and CBV infection was abrogated, without diminishing the replication of virus in the pancreas. Thus beta cell injury in DADM appears to be T cell-mediated.


Antibody response of children immunized with poliovaccines: an evaluation using two strains of poliovirus type 1. Neutralizing antibody response of children immunized with either OPV (3 doses), or IPV (2 doses) was evaluated against poliovirus type 1 Sabin vaccine strain and a local neurovirulent isolate. Both vaccines elicited significantly better antibody response against the vaccine strain than against the neurovirulent isolate. Moreover, approximately 35 per cent of sera contained very low levels of antibody against the virulent virus in spite of good antibody titre against the vaccine strain. The observed difference in antibody response to the wild and the vaccine strains was significant. The differential immune response could be one of the reasons of paralytic disease observed even after administration of OPV (3), in some children if infecting virus dose is high, as in case of urban slums in endemic areas.


Susceptibility of Aedes krombeini cell line to some arboviruses. The susceptibility of the newly established Ae. krombeini cell line (NIVI-AK-453) to six arboviruses, belonging to four different families, was studied. Sindbis (SIND), Vesicular stomatitis (VSV) Chandipura (CHP) and African horse sickness (AHS) viruses multiplied in these cultures. A four-to-five-fold increase in the virus titres was observed. The maximum titre of SIND, VSV, CHP and AHS viruses were observed on 1st, 4th, 3rd and 10th post infection days, respectively. A steady and significant increase in the titre of AHS was observed over a period of ten days. The sandfly fever virus (SFV) and the tick-borne, Kaisodi virus did not multiply in the cultures.