Proliferating cell nuclear antigen (PCNA) immunostaining as an alternative to bromodeoxyuridine (BrdU) immunostaining for brain tumours in paraffin embedded sections. Immunohistochemical staining for proliferating cell nuclear antigen (PCNA) and BrdU using anti-PCNA and anti-BrdU monoclonal antibodies, respectively, was performed in 16 human brain tumours, including 3 glioblastomas multiforme, 2 anaplastic astrocytomas, 1 cerebellar astrocytoma, 2 recurrent meningiomas, 4 non-recurrent meningiomas, 3 neurinomas and 1 medulloblastoma. Patients with brain tumours received an injection of bromodeoxyuridine (BrdU) intravenously during surgery, and tumour specimens were fixed in 70% ethanol and embedded in paraffin. The percentage of positive cells for PCNA was compared with a BrdU labelling index using adjacent paraffin-embedded sections. The percentage of PCNA-positive cells was correlated with the BrdU labelling index and the histological malignancy of the brain tumours. The correlation coefficient was 0.84. This suggests that the immunohistochemical staining for PCNA in paraffin sections is a good alternative to the BrdU labelling index.


Prevention of post-traumatic excitotoxic brain damage with NMDA antagonist drugs: a new strategy for the nineties. Excitotoxic mechanisms due to overactivity of the amino acid neurotransmitters glutamate and aspartate maybe responsible for brain damage after injury. In this review we examine ischaemia and shear injury, which are relevant to human head injury. The opportunities for treatment using glutamate antagonist drugs are discussed.


C-fos expression during electroacupuncture analgesia in rats--an immunohistochemical study. Although the central mechanisms of electroacupuncture analgesia (EAA) have been investigated, a systematic study for the involvement of neuronal populations of central nervous system (CNS) in EAA has not been well undertaken, largely due to the difficulty in tracing the neuronal pathways by traditional techniques. Recently developed c-fos expression examination by immunohistochemical method with Ab-1 antisera might be used for this purpose as a useful marker for neuronal activity in CNS. In this study, tail flick latency (TFL) was tested as an index of pain threshold in conscious rats. After unilateral electroacupuncture was applied at 'Zuan-san-li' and 'Huan-tiao', the TFL was significantly prolonged. To explore the possible involvement of certain neuronal groups of central nervous system in EAA, we examined the EAA accompanied c-fos expression throughout the neuraxis, and a lot of specific c-fos protein labelled neurons were found in lumbar spinal cord (laminae I and II), nucleus raphe magnus, nucleus raphe dorsalis, substantia grisea centralis, nucleus habenulae lateralis, nucleus habenulae medialis, nucleus medialis thalami, nucleus lateralis hypothalami, nucleus supramamillaris, nucleus supraopticus, nucleus arcuatus, nucleus preopticus medialis, nucleus amygdala, nucleus tractus diagonalis, etc. No obvious c-fos expression was shown in these areas on control rats. These results strongly suggested that the functional activation of above-mentioned nuclei by electroacupuncture was underlied in EAA action.


Acupuncture treatment for psoriasis: a retrospective case report. We treated 61 cases of psoriasis with acupuncture, including 25 patients with complications of joint involvement and two cases with scleroderma additionally. All of the patients had failed to respond to their prior conventional western medical management. 25 patients were males and 36 were females. Their ages ranged from 22 to 84 years, with an average of about 52 years. There was no significant difference of the average ages between the sexes. Most of them (about 61%) had quite extensive involvement of the body. The average of duration of their illness was over 16 years, ranging from two to 65 years. They received an average of about nine sessions of acupuncture treatment, ranging from one to 15. Almost one third (19) of them had eleven to thirteen sessions. With the acupuncture treatment, about one-half (30) of the 61 patients had complete or almost complete clearance of the skin lesions. About a quarter (14 patients) of them had a clearance of about two thirds of the skin lesions. Eight of them had a clearance of one third of the skin lesions. Nine patients had minimal or no improvement. Our experience indicates that acupuncture is induced an effective therapeutic modality for psoriasis, particularly when the western medical management is unsuccessful. We speculated about the possible involvement of the cutaneous reticuloendothelial system in the clearance of the skin lesions.


Bacterial polysaccharides, endotoxins, and immunomodulation. These studies show that at least some--though certainly not all--of the adjuvant effects of LPS and its derivatives can be attributed to its ability to eliminate the inhibitory effects of Ts which are activated during the course of a normal immune response. The ability of nontoxic MPL to act in this fashion suggests that it can be used as a safe and acceptable alternative to Freund's complete adjuvant to increase the immunogenicity of poorly immunogenic antigens. More important, the ability of MPL to eliminate the expression of Ts activity, without adversely influencing other T cell functions (e.g., Th, Ta, and Tc activity) makes its use as an adjuvant even more promising since it can then permit those T cell functions to be expressed in a much more efficient manner. Obviously, this would have great significance for the development of tumor immunity.


[Management of femoral neck fractures with the spongiosa screw and the dynamic hip screw]. Between 1983 and 1987 419 patients with femoral neck fractures were operated on at the University Clinics of Graz. In 120 patients we could preserve the head of the femur doing an internal fixation with cancellous bone screws or DHS. 68 of them we personally could control with a mean follow up of 45.2 months. The most important complications were the necrosis of the head which were observed in 19.1% of our patients and the non union which occurred in 5.8% but not all these patients required reoperation.


Hypothalamic paraventricular nucleus plays a role in acupuncture analgesia through the central nervous system in the rat. This work investigates the effect of hypothalamic paraventricular nucleus (PVN) on acupuncture analgesia in the rat. Electrical stimulation of PVN or injection of L-glutamate sodium into PVN could enhance the analgesic effect induced by acupuncture Zusanli (St. 36) while the electrical cauterization of PVN is decreased; Removal of the pituitary could not influence the effect of enhancing acupuncture analgesia induced by the injection of L-glutamate sodium into PVN. These results suggest that PVN might play an important role in acupuncture analgesia through central nervous system.


[Management of unstable pertrochanteric and per- to subtrochanteric femoral fractures with the dynamic hip screw]. The operative treatment of fractures of the proximal end of femur with the dynamic hip screw (DHS) permits a weight-bearing stable osteosynthesis. In 5.2 years between 1985 and 1990 531 patients were treated with a DHS. The average age was 75 years. This method of osteosynthesis was also performed in 59 patients with unstable per- or per-subtrochanteric fractures. Complications such as pseudarthrosis, necrosis or penetration of the head of femur, implant bending or breaking were not observed. The dynamic hip screw is a valid procedure for an early and weight-bearing treatment also for unstable pertrochanteric fractures of the femur with special benefits for the elderly.


[Results of angled plated osteosynthesis in per- and subtrochanteric fractures in the elderly]. From 1/86 to 12/89 62 old patients (average age: 82.6) with intertrochanteric and subtrochanteric fractures underwent surgery by using angeled plates. The postoperative course showed complications in 32.2%, while only 1 patient died. In 3 of 5 cases with dislocation of the angeled plate re-osteosynthesis was performed. 4 other complications (2x haematoma, 2x wound dehiscence) needed further surgery. During the first three months 3 other dislocations occurred. In two cases re-osteosynthesis was carried out. Due to these results and the high rate of complications we are of the opinion that the indication for the use of the angeled plate osteosynthesis in old patients has to be limited in favor of other methods like the dynamic hip screw or total hip replacement.


Improvement of motor function in multiple sclerosis by use of protopam chloride. PAM, a cholinesterase reactivator, was administered orally and parenterally to 37 patients with multiple sclerosis and a control group of 24 patients with other neurological diseases and pain syndromes. The effects of the administration of this compound in these patients with and without electrical stimulation of the spinal cord were studied. The clinical response to the drug follows a known time course and is dose related. Administration of large doses orally or intravenously aggravates existing neurological dysfunction. With a dose of 1,000 mg intravenously, a characteristic response is the temporary appearance of new ophthalmological abnormalities, followed by significant improvement in motor control and behavior, which gradually subsides. Parenteral administration is superior to oral. Tolerance to the drug is observed. The presence of electrical stimulation of the spinal cord complements the action of the drug. When electrical stimulation is withdrawn, the effect of the drug reproduces the effect of the electrical stimulation. It is suggested there is a defect in cholinesterase in multiple sclerosis patients, and its reactivation may have a significant relationship to signs and symptoms.


Frequency-dependent effects of sine-wave cranial transcutaneous electrical nerve stimulation in human subjects. In a double-blind protocol, ninety healthy volunteer subjects received 30 minutes of constant current sine-wave cranial transcutaneous electrical nerve stimulation (TENS) of 5 Hertz (Hz), 100 Hz, or 2000 Hz frequency (current maintained below .5 mA for safety), placebo TENS, or no treatment. The five groups were compared on pre- to posttreatment changes in blood pressure, heart rate, peripheral temperature, and anxiety. Analysis showed significant reductions in systolic and diastolic blood pressure and heart rate after 100 Hz cranial TENS as compared to the other groups. No other differences achieved significance.


[The value of Ender nailing in hip para-articular femoral fractures in gerontologic traumatology. Ender nailing--hip para-articular femoral fractures--gerontologic traumatology]. Nailing according to Ender is one of several competing methods of osteosynthesis for treating per- and subtrochanteric fractures of the femur in geronto-traumatology. Within 5 years we have treated 62 geriatric patients by this procedure. The average-age of our patients was about 76 years. Only 20 of them were able to be examined after a time of ca. 37 months post operationem. Intraoperative complications were backing out of the femur corticalis at the entry hole, followed by perforations of the head or neck of the femur. In one case we saw a supracondylar fracture. In the postoperative period we mostly found dislocations of the nails to cranial or caudal. Moreover there were one wound infect and one pseudarthrosis. The mortality rate came to 6.5% and was never caused by the method. When leaving the hospital the majority of the elderly patients was mobilized and able to walk. In spite of reduction of function of hip and knee and external rotation deformity and shortened legs the patients declared to be content with the result of the operation. Important geronto-traumatological aspects of the Endernailing-method could be seen in the simple procedure, the short operating time, a minimal surgical trauma and a diminishing of the risk of the mostly multi-morbid patients. Regarding the great number of specific complications competing methods are going on to be preferred. Meanwhile the application of Endernails is an exception in geronto-traumatology.


[Alloplastic replacement of the anterior cruciate ligament. Material technical and biomechanical principles, surgical technique]. The Trevira-ligament of Polyaethylenter-ephthalat (= Trevira-hochfest 730) seems to be the best qualified replacement of the anterior cruciate ligament at the present. It meets all material and technical requirements and makes allowances for all biomechanical knowledges. The modified over-the-top-technique by arthroscopy or mini-arthrotomy (minimal operative trauma) makes it possible to conserve still existing ligamentous and soft tissue and guarantees the post-operative immediate functional therapy. A potential reinstability--independent of each cause--doesn't imply worse starting conditions and enables corrections including the substitute of a ruptured synthetic ligament.


[Tarsal tunnel syndrome after fibular ligament repair. A rare complication of surgical therapy of chronic instability of the ankle joint ligament]. A case of tarsal tunnel syndrome following a reconstructive procedure with repair of chronic ankle instability using periosteal flap is reported. Two pathogenetic factors have to be discussed as possible causes for this complication. Dissection of the medial retinaculum was performed. With this procedure the patient is complaints could be removed.


Development and inhibition of mouse ear oedema induced with capsaicin. Oedema was induced in one ear of male mice of the CFLP strain with capsaicin solution (10 microliters/40 micrograms/ear). The development in time and the extent of the oedema were determined by the oedema-disk gravimetric technique. The maximum oedema was attained in less than 1 h, and there was, subsequently, a gradual decrease. The extent of the mouse ear oedema induced in this way and measured after 60 min was inhibited to a statistically significant degree and in a dose-dependent manner by the antihistamine chloropyramine, the antihistamine-antiserotonin cyproheptadine, the non-steroidal anti-inflammatory agent piroxicam, the prostaglandin antagonist di-4-phloretin phosphate, and the lipoxygenase inhibitor nordihydroguairetic acid. The method proved suitable for the detection of oedema and for the biologically quantitative determination of the state of desensitization induced with capsaicin.


["In-dartos" orchidopexy in the treatment of cryptorchism]. Undescended testis is a common congenital abnormality. The main complications are infertility and venue of malignant testicular tumor. If early orchiopexy can prevent infertility, it remains a high risk of venue of a malignant tumor. So, the best technique of orchiopexy is that which allows an easy clinical follow-up of the undescended testis after surgical treatment, and doesn't involv the hematoseminal barrier. From the analysis of fourty cases of scrotal pouch orchiopexy and a review of the literature, it appears that this procedure is efficient and gives more than 90% of good results.


[Neurotransmission and the lower urinary system]. Various substances, called neurotransmitters, act as a chemical relay between the nervous and the tissue of the lower urinary tract. Some play also a role inside the muscle cell modulating the activity of the myofibrils. To allow a better pharmacochemical approach, these substances are described, their activity analyzed, with the purpose of helping specialists concerned by investigation or treatment of the lower urinary tract, to be able to act on their activity or change their pathologic abnormalities.


[Effect of YM-12617 (amsulosin hydrochloride) on lower urinary tract function in the female decerebrate dog]. The effect of YM-12617 on the lower urinary tract function was studied by combined recording of cystometry and external sphincter electromyogram (EMG) in 11 female decerebrate dogs. Reflex micturitions were induced by bladder filling before and after YM-12617 administration. Statistical analysis was carried out on the urodynamic parameters. YM-12617 in a dose of 10 micrograms/kg significantly decreased micturition threshold pressure during the collecting phase. In the urodynamic parameters of the emptying phase there was a significant decrease in contraction pressure at 10 and 30 micrograms/kg.


[Prader-Willi syndrome associated with chromosomal aberration: report of a case]. A male case of Prader-Willi syndrome (2.8 years in age) with an interstitial deletion of a chromosome affecting 15q 11-12 region is reported. The chief complaints were hypoplastic scrotum and defect of bilateral scrotal content. The clinical features were short stature, obesity, delayed mental development, bilateral cryptorchidism, hypogenitalism, hypopigmentation, and bilateral moderate vesicoureteral reflux with a history of muscular hypotonia. Bilateral orchidopexy was done. Endocrinologically both base values of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were normal although LH reserve function was impaired on gonadotropin releasing hormone (GnRH) test. Testosterone response was normal by the stimulation of human chorionic gonadotropin. An interstitial deletion of proximal 15q, and pituitary-gonadal axis in Prader-Willi syndrome are discussed in relation to the clinical features and therapy.


National guidelines for the management of asthma in adults. Asthma is now considered primarily an inflammatory disease in which bronchospasm occurs secondary to airway inflammation. Management strategies include the use of inhaled anti-inflammatory agents, notably inhaled corticosteroids and cromolyn. Mild intermittent asthma may be treated with inhaled bronchodilators. Moderate asthma should be treated with an inhaled anti-inflammatory agent in addition to an inhaled beta agonist. If symptoms persist, an oral bronchodilator (either a beta-adrenergic agonist or theophylline) should be added. Therapy for severe asthma includes combinations of the foregoing medications, with the possible addition of oral corticosteroids. Other aspects of management include the use of a spacer device with inhaler therapy, control of concomitant allergies and triggering factors such as chronic sinusitis, tobacco smoke and gastroesophageal reflux, and home use of a portable peak flow meter to monitor the disease.


Autocrine and paracrine mechanisms in the pathophysiology of heart failure. At the cellular and molecular level the transition to heart failure is a complex process that involves structural adaptation, not only of the heart, but of peripheral vasculature and renal tissues as well. Recent studies have suggested that autocrine, paracrine, and circulating biologically active mediators activate events that result in the concerted failure of adaptive mechanisms and the ultimate depression of cardiac myocyte function. Greater understanding of these local mechanisms in the future may lead to drug therapies that can selectively block these mechanisms and prevent the progression from compensation to overt heart failure.


Beta-blocking effect of propafenone based on spectral analysis of heart rate variability. RR variability was analyzed in 15 patients with ventricular arrhythmias to evaluate whether the antiarrhythmic action of propafenone is associated with alteration of neural control mechanisms. Before drug administration, spectral analysis of RR variability was characterized by 2 major components at low and high frequency, which are considered to reflect sympathetic and parasympathetic modulation of the heart period. After propafenone (600 to 900 mg/day), there was a marked reduction in RR variance (826 +/- 184 to 412 +/- 77 ms2; p < 0.05), although the mean RR interval was unchanged. The drug significantly reduced the low-frequency component (52 +/- 6 to 28 +/- 4 nu) and augmented the high-frequency component (39 +/- 6 to 55 +/- 5 nu). As a result, the low-/high-frequency ratio (an index of sympathovagal balance) decreased from 2.0 +/- 0.4 to 0.6 +/- 0.1. A positive correlation between serum levels and drug-induced changes in the low-frequency component was also observed. Furthermore, the increase in the low-frequency component induced by tilt (53 +/- 5 to 79 +/- 3 nu) was markedly attenuated after drug administration (27 +/- 5 to 54 +/- 7 nu). Thus, propafenone administration is associated with changes in spectral components that are consistent with a beta-blocking effect of the drug.


Protection against histamine-induced bronchoconstriction by loratadine. In a simple blind pilot study we have examined the protective effect of loratadine, a new H1 histamine receptor antagonist, against bronchoconstriction induced by histamine inhalation. Six patients with an episodic or continuing obstructive bronchial disease and proven bronchial hyperreactivity were submitted to two identical histamine challenges, first without premedication and then after three days' treatment with 10 mg loratadine daily. The mean FEV1 fall in the first test after histamine inhalation was 33% and in the second test after pretreatment with loratadine only 3.3% (p less than 0.01). Five out of six patients have shown complete protection against histamine-induced bronchoconstriction, in one patient the protection was partial. We conclude that loratadine effectively inhibits histamine-induced bronchoconstriction and could be useful in the prophylactic treatment of asthma and obstructive bronchitis.


Early results with bilateral internal mammary artery grafting in coronary reoperations. In recent years, use of the internal mammary artery (IMA) as first graft of choice has been expanded with bilateral and sequential grafts in primary myocardial revascularization. The use of bilateral IMA grafts in reoperation has seldom been reported. The experience and early results with bilateral IMA grafting in 47 patients undergoing coronary reoperation are described. Hospital mortality was 6.3%. Four patients had postoperative signs of low cardiac output, and 4 had a perioperative myocardial infarction. At follow-up (18 +/- 18 months), 2 cardiac-related, late deaths were noted. Thirteen patients (29%) improved 1 New York Heart Association class, and 28 (63%) improved > 1 class. In 1 of 44 surviving patients, operation did not result in a decrease in angina. On the basis of the early results, the bilateral use of the IMA in coronary reoperation appears justified.


p53 and c-erbB-2 protein expression in breast carcinomas. An immunohistochemical study including correlations with receptor status, proliferation markers, and clinical stage in human breast cancer. Receptor status, proliferative activity, loss of differentiation, inactivation of tumor suppressor genes, and overexpression of oncogenes are related events that may affect the prognosis of patients with breast cancer. Ninety-seven unselected breast carcinomas were immunostained for estrogen and progesterone receptors, Ki-67 proliferation-associated antigen, p53 tumor suppressor gene product (p53), and c-erbB-2 protein. Immunohistochemical results and clinical data were compared. Altered p53 expression (regarded as indirect indication of inactivating gene alterations) was found in 25.8% of cases and was associated with a high Ki-67 labeling index, high mitotic count, and high histologic grade, with c-erbB-2 overexpression, and with negative estrogen and progesterone receptor status. p53 immunostaining could be found also in cytologic samples and correlated with p53 immunoreactivity on frozen sections of the corresponding tumors. c-erbB-2 protein overexpression was seen in 24.7% of cases and was associated with p53 altered expression and negative receptor status. Double immunohistochemical staining showed p53 and c-erbB-2 immunoreactivity in the same cells. Median and mean +/- standard deviation Ki-67 labeling index values were 15 and 16.32 +/- 10.05, respectively. Ki-67 labeling index was correlated with high mitotic count and was positively associated with histologic grade, negative progesterone receptor status, and p53 expression. Estrogen receptor status was not associated with any histologic or clinical parameters, whereas progesterone receptor status was associated with grading. The direct relation of p53 protein alterations with c-erbB-2 overexpression may be interpreted in light of the multistep model of tumor progression. Cases with altered expression of both p53 and c-erbB-2 proteins could be interpreted as having lost one inhibitory control mechanism of cell proliferation and having gained one activator of the malignant potential. However, in comparing cases with the p53 + c-erbB-2 + phenotype with cases showing positivity for only one of these gene products, no association with higher stages was seen. Detection of p53 altered expression on cytologic samples of malignant tumors may have diagnostic relevance, and p53 immunostaining may prove to be an additional diagnostic criterion in cytologic diagnosis.


Chronic 3,4-methylenedioxymethamphetamine (MDMA) use: effects on mood and neuropsychological function? 3,4-Methylenedioxymethamphetamine (MDMA; "ecstasy") is a selective serotonin (5-HT) neurotoxin in animals. There is now preliminary evidence in humans of 5-HT deficits associated with extensive use of MDMA. In order to begin to describe the cognitive and mood effects of chronic MDMA use, nine individuals with extensive MDMA use histories were studied. Despite the absence of memory deficits on clinical examination, a pattern of mild-to-moderate impairment was observed on both the Initial and Delayed Paragraph Tests of the Wechsler Memory Scale; eight of the subjects had at least mild impairment on at least one test in the neuropsychological battery. Despite previously reported suggestive evidence of 5-HT deficit in this group, none reported depressed mood or met clinical criteria for an affective disorder at the time of testing. These preliminary findings raise concern about possible detrimental effects of MDMA use on neuropsychological function for future systematic study and they highlight important issues regarding the effects of 5-HT deficits on cognitive function and mood regulation.


Menopause and serum cholesterol: differences between blacks and whites. The Minnesota Heart Survey. The relation between menopause and serum total and high-density-lipoprotein cholesterol was examined by the Minnesota Heart Survey in a cross-sectional, population-based study of 344 black women and 474 white women aged 35-54 years from the Twin Cities metropolitan area in 1985-1986. Analysis of covariance was used to examine differences in serum total and high-density-lipoprotein cholesterol in black women and white women by menopausal status, adjusting for the effects of age, educational level, cigarette smoking, body mass index, exercise, alcohol consumption, diabetes mellitus, sex hormone, beta blocker, and diuretic use. Among whites, adjusted serum total cholesterol was 13 mg/dl higher in postmenopausal than in premenopausal women (p less than 0.002). Black postmenopausal women had slightly higher serum total cholesterol than did their premenopausal counterparts (5.4 mg/dl). However, this was not statistically significant. An interaction term in a linear regression model confirmed a racial difference in the total cholesterol association with menopause (p less than 0.02). The higher total cholesterol levels observed in white postmenopausal women were mainly among those with natural menopause (20.7 mg/dl higher than premenopausal, p less than 0.0003) and those with a hysterectomy and at least one intact ovary (11.0 mg/dl higher, p = 0.05). Among black women, only the subgroup with a hysterectomy and a bilateral oophorectomy had a significantly higher serum total cholesterol (19.9 mg/dl higher than premenopausal, p less than 0.05). There was no significant association between high-density-lipoprotein cholesterol and any type of menopause in either black women or white women. Our findings may reflect a true physiologic difference in the relation between menopause and serum total cholesterol between American blacks and whites. The lack of a significant association between menopause and high-density-lipoprotein cholesterol in either race raises the possibility that menopause may not affect atherosclerosis risk via reduced high-density-lipoprotein cholesterol.


Seroepidemiology of California and Bunyamwera serogroup bunyavirus infections in humans in California. Several human populations in California were surveyed cross-sectionally and longitudinally for neutralizing antibodies to selected arthropod-borne bunyaviruses in the California and Bunyamwera serogroups. Overall, the prevalence of antibodies to California serogroup viruses was 6.4% in 702 individuals sampled during 1963-1988. Comparative antibody titers in individual sera indicated that 4.1% and 1.6% of these infections were caused by viruses similar or identical to Jamestown Canyon and California encephalitis, respectively. Evidence of prior infection with the Jamestown Canyon serotype was found in 10% of 118 humans employed outdoors in high elevation areas and sampled in 1988, including 5 of 16 persons (31%) employed as rangers patrolling in remote forests and meadows. This probably reflects increased exposure to bites of boreal mosquitoes that breed in pools of melted snow. Antibodies to Bunyamwera serogroup viruses, including the Northway serotype, which was recently shown to be enzootic in California, were found in only 2 of 702 humans studied. No seroconversions were detected to selected California or Bunyamwera serogroup viruses in paired samples from 392 humans, including 349 patients with acute central nervous system disease or undifferentiated febrile illnesses who were sampled during 1963-1988, and thus these viruses are currently unconfirmed as human pathogens in California.


Symptomatic La Crosse virus infections of the central nervous system: a study of risk factors in an endemic area. In most years, La Crosse virus is the most common cause of reported mosquito-borne illness in the United States. The authors conducted a case-control study to determine if behavioral and environmental factors influenced the risk of La Crosse virus illness. Data were gathered on 31 serologically confirmed cases and 60 age-, sex-, and geography-matched controls in West Virginia in 1987 and 1988. Univariate analysis revealed minimal elevation of disease risk (odds ratios (ORs) less than 2.0) with increased time outdoors, non-use of insect repellent, non-use of air conditioning, lack of screened windows, and not wearing protective clothing. Univariate and multivariate analysis indicated that the presence of tree holes significantly increased disease risk (OR = 8.5 for greater than or equal to 1 tree hole vs. 0 tree holes). The following factors may also increase disease risk, although the findings were not statistically significant: discarded tires (OR = 3.2 for greater than or equal to 10 tires vs. 0-9 tires); non-tire artificial containers (OR = 4.1 for greater than or equal to 6 containers vs. 0-5 containers); and close proximity of the house to the forest edge (OR = 3.2 for 0-49 ft (0-14.9 m) vs. greater than or equal to 50 ft (greater than or equal to 14.9 m)). The authors conclude that the presence of natural breeding sites (tree holes) is an important risk factor for La Crosse virus illness. These results may be important in guiding future efforts aimed at preventing infection with La Crosse virus.


Trisomy 15 with loss of the paternal 15 as a cause of Prader-Willi syndrome due to maternal disomy. Uniparental disomy has recently been recognized to cause human disorders, including Prader-Willi syndrome (PWS). We describe a particularly instructive case which raises important issues concerning the mechanisms producing uniparental disomy and whose evaluation provides evidence that trisomy may precede uniparental disomy in a fetus. Chorionic villus sampling performed for advanced maternal age revealed trisomy 15 in all direct and cultured cells, though the fetus appeared normal. Chromosome analysis of amniocytes obtained at 15 wk was normal in over 100 cells studied. The child was hypotonic at birth, and high-resolution banding failed to reveal the deletion of 15q11-13, a deletion which is found in 50%-70% of patients with PWS. Over time, typical features of PWS developed. Molecular genetic analysis using probes for chromosome 15 revealed maternal disomy. Maternal nondisjunction with fertilization of a disomic egg by a normal sperm, followed by loss of the paternal 15, is a likely cause of confined placental mosaicism and uniparental disomy in this case of PWS, and advanced maternal age may be a predisposing factor.


High-resolution mapping of the X-linked hypohidrotic ectodermal dysplasia (EDA) locus. The X-linked hypohidrotic ectodermal dysplasia (EDA) locus has been previously localized to the subchromosomal region Xq11-q21.1. We have extended our previous linkage studies and analyzed linkage between the EDA locus and 10 marker loci, including five new loci, in 41 families. Four of the marker loci showed no recombination with the EDA locus, and six other loci were also linked to the EDA locus with recombination fractions of .009-.075. Multipoint analyses gave support to the placement of the PGK1P1 locus proximal to the EDA locus and the DXS453 and PGK1 loci distal to EDA. Further ordering of the loci could be inferred from a human/rodent somatic cell hybrid derived from an affected female with EDA and an X;9 translocation and from studies of an affected male with EDA and a submicroscopic deletion. Three of the proximal marker loci, which showed no recombination with the EDA locus, when used in combination, were informative in 92% of females. The closely linked flanking polymorphic loci DXS339 and DXS453 had heterozygosities of 72% and 76%, respectively, and when used jointly, they were doubly informative in 52% of females. The human DXS732 locus was defined by a conserved mouse probe pcos169E/4 (DXCrc169 locus) that cosegregates with the mouse tabby (Ta) locus, a potential homologue to the EDA locus. The absence of recombination between EDA and the DXS732 locus lends support to the hypothesis that the DXCrc169 locus in the mouse and the DXS732 locus in humans may contain candidate sequences for the Ta and EDA genes, respectively.


DNA polymorphism of alkaline phosphatase isozyme genes: linkage disequilibria between placental and germ-cell alkaline phosphatase alleles. The use of human placental alkaline phosphatase (PLAP) cDNA as a probe allows the detection and identification of restriction DNA fragments derived from three homologous genes, i.e., intestinal alkaline phosphatase (AP), germ-cell AP (GCAP), and PLAP. In previous RFLP studies we have reported linkage disequilibria between an RsaI and two PstI (a and b) polymorphic restriction sites and electrophoretic types of PLAP. In this report we present evidence that, in spite of the strong correlation with PLAP types, PstI(b) is an RFLP of GCAP. The data indicate close linkage between the PLAP and GCAP loci.


X-linked nephrogenic diabetes insipidus: from the ship Hopewell to RFLP studies. Nephrogenic diabetes insipidus (NDI; designated 304800 in Mendelian Inheritance in Man) is an X-linked disorder with abnormal renal and extrarenal V2 vasopressin receptor responses. The mutant gene has been mapped to Xq28 by analysis of RFLPs, and tight linkage between DXS52 and NDI has been reported. In 1969, Bode and Crawford proposed, under the term "the Hopewell hypothesis," that most cases in North America could be traced to descendants of Ulster Scots who arrived in Nova Scotia in 1761 on the ship Hopewell. They also suggested a link between this family and a large Mormon pedigree. DNA samples obtained from 13 independent affected families, including 42 members of the Hopewell and Mormon pedigrees, were analyzed with probes in the Xq28 region. Genealogical reconstructions were performed. Linkage between NDI and DXS304 (probe U6:2.spl), DXS305 (St35-691), DXS52 (St14-1), DXS15 (DX13), and F8C (F814) showed no recombination in 12 families, with a maximum lod score of 13.5 for DXS52. A recombinant between NDI and DXS304, DXS305, was identified in one family. The haplotype segregating with the disease in the Hopewell pedigree was not shared by other North American families. PCR analysis of the St14 VNTR allowed the distinction of two alleles that were not distinguishable by Southern analysis. Carrier status was predicted in 24 of 26 at-risk females. The Hopewell hypothesis cannot explain the origin of NDI in many of the North American families, since they have no apparent relationship with the Hopewell early settlers, either by haplotype or by genealogical analysis. We confirm the locus homogeneity of the disease by linkage analysis in ethnically diverse families. PCR analysis of the DXS52 VNTR in NDI families is very useful for carrier testing and presymptomatic diagnosis, which can prevent the first manifestations of dehydration.


Linkage disequilibrium among RFLPs at the insulin-receptor locus despite intervening Alu repeat sequences. Multiple mutations of the insulin receptor (INSR) gene have been identified in individuals with extreme insulin resistance. These mutations have included recombination events between Alu repeat units in the tyrosine kinase-encoding beta-chain region of the gene. To evaluate the influence of Alu and dinucleotide repetitive sequences on recombination events within the insulin receptor gene, I examined the degree of linkage disequilibrium between RFLP pairs spanning the gene. I established 228 independent haplotypes for seven RFLPs (two each for PstI, RsaI, and SstI and one for MspI and 172 independent haplotypes which included an additional RFLP with BglII) from 19 pedigrees. These RFLPs span > 130 kb of this gene, and my colleagues and I previously demonstrated that multiple Alu sequences separate RFLP pairs. Observed haplotype frequencies deviated significantly from those predicted. Pairwise analysis of RFLP showed high levels of linkage disequilibrium among RFLP in the beta-chain region of the insulin receptor, but not between alpha-chain RFLPs and those of the beta-chain. Disequilibrium was present among beta-chain RFLPs, despite separation by one or more Alu repeat sequences. The very strong linkage disequilibrium which was present in sizable regions of the INSR gene despite the presence of both Alu and microsatellite repeats suggested that these regions do not have a major impact on recombinations at this locus.


Helicobacter pylori seroprevalence in patients with rheumatoid arthritis: effect of nonsteroidal anti-inflammatory drugs and gold compounds. To investigate the relationship between Helicobacter pylori infection and nonsteroidal anti-inflammatory drug (NSAID) intolerance and the effect of gold use on the seroprevalence of H. pylori. We examined the frequency of discontinuation of NSAIDs in 132 unselected patients with rheumatoid arthritis attending an outpatient subspecialty clinic, and the effect of gold compound use on the seroprevalence (by IgG enzyme-linked immunosorbent assay) of H. pylori infection in this population. Logistic and multivariate regression analysis was performed adjusting for age, gender, ethnic origin, history of ulcer, and duration of rheumatoid arthritis. Fifty-four patients had a positive serology for H. pylori (41%). Twenty-seven of the seropositive patients (50%), versus 45 of the seronegative patients (57.7%), had to discontinue NSAIDs (aspirin and/or nonaspirin) at least once since their diagnosis of rheumatoid arthritis because of gastrointestinal side effects (odds ratio [OR], 0.93; 95% confidence interval [CI], 0.63 to 1.38). Forty-one of the seropositive patients (76%) had received gold compounds as compared with 62 of the seronegative patients (79.5%) (OR: 0.96; 95% CI: 0.61 to 1.50). We did not find any relationship between H. pylori seropositivity and NSAID intolerance in patients with rheumatoid arthritis. In addition, our results do not demonstrate a reduction in H. pylori seroprevalence in rheumatoid arthritis patients treated with gold compounds.


Parental origin determination in thirty de novo Robertsonian translocations. Cytogenetic heteromorphisms and restriction fragment length polymorphisms were used to assign the parental origins of 30 de novo non-homologous Robertsonian translocations. The balanced and unbalanced translocations studied included 20 rob(14q21q) four rob(13q14q)four rob(15q21q) one rob(13q15q), and one rob(13q21q). Significantly more maternally (26/30) than paternally (4/30) derived de novo translocations were noted and all rob(14q21q) ascertained through unbalanced probands (20/20) were maternal in origin. Interestingly, 12/13 probands who were trisomic and informative for proximal chromosome 21q loci were homozygous for the markers tested. Segregation (2:1) of the Robertsonian translocation into one daughter cell in meiosis I and subsequent failure of the chromosome 21 chromatids to separate in meiosis II may account for our observation of homozygosity for proximal chromosome 21 loci in the majority of de novo rearrangements tested.


Diagnosis of arylsulfatase A deficiency. Metachromatic leukodystrophy (MLD) is a neurologically devastating autosomal recessive disorder in humans associated with deficient arylsulfatase A activity. However, clinically normal individuals described as being pseudo-arylsulfatase-A deficient also demonstrate the same deficiency. Genotypically, they may be homozygous for the pseudodeficiency mutation (associated with 2 A-->G transitions in the cDNA of arylsulfatase A) or heterozygous with one pseudodeficiency and one MLD allele. Using as examples 2 families in which the pseudo deficiency condition occurs either independently or together with MLD, we demonstrate the utility of a proposed diagnostic protocol to provide complete genotype identification of individuals suffering from arylsulfatase A deficiency. Patient fibroblasts are extracted for DNA and a cytoplasmic fraction, which is used for arylsulfatase A enzyme assay. This will identify an arylsulfatase A-deficient group, which is further analyzed electrophoretically. Cells from the clinically affected patients with MLD are completely deficient in arylsulfatase A activity, whereas those from the pseudodeficient individuals demonstrate a characteristic residual arylsulfatase A activity detectable only after electrophoresis. Within this pseudodeficient group, gene amplification of DNA specific for the A-->G mutations will distinguish between those who are homozygous for the pseudodeficiency allele and those who are compound heterozygous for the pseudodeficiency and MLD alleles. This protocol of complete genotype identification requires only about 10(6) fibroblasts (1 x 100 mm dish) and 2 days to complete. Such variant-specific genotype identification increases accuracy and prognostic value of the diagnosis. It will likely become the preferred choice for diagnosis of genetic disease in the future as more variant-specific mutations are identified at the molecular level.


Hemorrhagic fever with renal syndrome in an endemic area of Greece. From 1983 to 1990, 32 patients with hemorrhagic fever with renal syndrome (HFRS) were admitted to our hospital. The diagnosis was confirmed by high IgM type titers of antibodies to Hantaan virus. All patients presented with serum and urine abnormalities suggesting renal involvement. Serum creatinine was elevated and ranged between 1.8 and 14.3 mg/dl. Proteinuria ranged between 0.5 and 6.4 g/24 h. Seven patients died due to shock or hemorrhage, while 6 patients were supported by hemodialysis or peritoneal dialysis. Five of them had a complete recovery. Two patients were discharged with some degree of renal impairment which remained stable 12-15 months later. Kidney biopsy in the first patient performed 1 year after his discharge revealed some degree of interstitial fibrosis and tubular atrophy as well as an area with ischemic and sclerosed glomeruli. We conclude that HFRS in Greece is a severe disease with a high mortality rate. The disease may cause chronic renal failure in a limited number of patients.


The relationship of quantitative nuclear morphology to molecular genetic alterations in the adenoma-carcinoma sequence of the large bowel. The relationship of abnormal nuclear morphology to molecular genetic alterations that are important in colorectal tumorigenesis is unknown. Therefore, Feulgen-stained isolated nuclei from 22 adenomas and 42 carcinomas that had been analyzed for ras gene mutations and allelic deletions on chromosomes 5q, 18q, and 17p were characterized by computerized image analysis. Both nuclear area and the nuclear shape factor representing irregularity correlated with adenoma-carcinoma progression (r = 0.57 and r = 0.52, P < 0.0001), whereas standard nuclear texture, a parameter of chromatin homogeneity, was inversely correlated with progression (r = -0.80, P < 0.0001). The nuclear parameters were strongly interrelated (P < 0.0005). In multivariate analysis, the nuclear parameters were predominantly associated with adenoma-carcinoma progression (P < or = 0.0001) and were not influenced significantly by the individual molecular genetic alterations. Nuclear texture, however, was inversely correlated with fractional allelic loss, a global measure of genetic changes, in carcinomas (r = -0.39, P = 0.011). The findings indicate that nuclear morphology in colorectal neoplasms is strongly related to tumor progression. Nuclear morphology and biologic behavior appear to be influenced by accumulated alterations in cancer-associated genes.


Thrombomodulin expression in malignant pleural mesothelioma and pulmonary adenocarcinoma. Thrombomodulin (TM) is a glycoprotein of molecular weight 75,000 kd that is normally present in restricted numbers of cells, including endothelial and mesothelial cells. In this study, the authors tested the possibility of using anti-TM to facilitate the diagnosis of mesothelioma. All of the 31 mesotheliomas and the two mesothelioma cell lines (MS-1 and MS-2) tested were stained positively with anti-TM. The specificity of anti-TM staining in mesothelioma cells was further confirmed by in situ hybridization of MS-1 cells with a TM-specific probe. The expression of TM in MS-1 cells was increased markedly when these cells were induced by 12-0-tetradecanyl phorbol 13-acetate (TPA) to differentiate. The expression of TM in mesothelioma cells, however, did not correlate with any particular phase of the cell cycle. In an attempt to differentiate pleural mesothelioma from pulmonary adenocarcinoma, the authors compared the expression of TM, carcinoembryonic antigen (CEA), and Leu M1 in these two types of tumors. Only four of 48 (8%) pulmonary adenocarcinomas were stained positively by antibodies to TM. Therefore, immunohistochemical staining with antibodies to TM yielded 100% sensitivity and 92% specificity for diagnosis of mesothelioma. All of the mesotheliomas stained negatively for CEA and Leu M1, except for one, which showed minimal focal positivity for Leu M1. In contrast, 79% and 60% of adenocarcinomas stained positively for CEA and Leu M1, respectively. These findings suggest that immunocytochemical staining with anti-TM should be added to the battery of tests to increase the diagnostic sensitivity and specificity for differentiating mesothelioma from pulmonary adenocarcinoma.


On the determination of turnover in vivo with tracers. Theoretical and practical aspects of the application of tracer methods for the measurement of turnover of blood-borne compounds are discussed, with special regard to lactate. The validity of the application of the tracer into the aortic arch and sampling from the right atrium (A-V), the administration of tracer into the vena cava and sampling from the aorta (V-A), and sampling to the determination of turnover are examined, using numerical examples. It is shown that the difference between specific activity in arterial and mixed venous blood depends mainly on the cardiac output, the ratio of tracee turnover to the mass of circulating tracee, and the sites of production and utilization of the tracee. Conditions are shown under which the A-V and V-A modes overestimate or underestimate the true rate of turnover. In theory, the A-V mode provides an exact estimate of turnover when the mean specific activity of the tracee in the whole body equals the specific activity of mixed venous blood in the right heart. It is shown that, for compounds with a high turnover rate, the underestimate in the A-V mode is small, and the mode provides a close approximation of true turnover. The underestimate in the V-A mode at high turnover rates is extensive. Experimental evidence indicates that, in several animal species, the specific activity of lactate and several amino acids in several organs and tissues nearly equals that in the venous blood, with the A-V mode providing a close approximation of the true turnover for these compounds.(ABSTRACT TRUNCATED AT 250 WORDS)


Cholecystokinin inhibits gastric acid secretion through type "A" cholecystokinin receptors and somatostatin in rats. The purpose of this study was to determine whether selective antagonism of type "A" cholecystokinin (CCK) receptors blocks inhibition of gastric acid secretion produced by CCK and whether this inhibition is mediated through either a somatostatin-dependent pathway or a vago-vagal reflex. Intravenous infusion of CCK (0.04-10 nmol.kg-1.h-1) dose dependently inhibited pentagastrin-stimulated gastric acid secretion in urethan-anesthetized rats, with a 50% inhibitory dose of 0.9 nmol.kg-1.h-1 and a maximum inhibition of approximately 50%. Blockade of type A CCK receptors using the selective type A receptor antagonist MK-329 completely reversed the inhibitory effect produced by a maximal dose (4 nmol.kg-1.h-1) of CCK. Immunoneutralization of endogenous somatostatin by administration of somatostatin monoclonal antibody abolished the inhibition produced by CCK. Concentrations of somatostatin in portal venous plasma were significantly increased after CCK administration; the increase in somatostatin was blocked by pretreatment with MK-329. In contrast, CCK-induced inhibition of gastric acid secretion was unaltered after perivagal capsaicin treatment. These results indicate that CCK inhibits gastric acid secretion in rats by activation of type A CCK receptors and through release of endogenous somatostatin.


Ascending contraction mediated by 5-hydroxytryptamine3 receptors in canine small intestine. We investigated the mechanism of ascending contraction induced by activation of 5-hydroxytryptamine3 (5-HT3) receptors in anesthetized dogs. Pressure-measuring balloons were inserted into a loop of extrinsically denervated jejunum. Drugs were administered via the arterial tree to the oral or the anal segment and the ensuing mechanical responses were monitored. Administration of 2-methyl-5-HT (440 pmol-44 nmol) to the anal segment caused contractions in the oral segment in a dose-dependent manner. This response was inhibited by treating the anal segment with ICS 205-930, cocaine, hexamethonium, or tetrodotoxin and by treating the oral segment with atropine or hexamethonium. The response persisted even after abolition of contraction in the anal segment by nifedipine. These results imply that activation of 5-HT3 receptors can induce an ascending contraction through an enteric excitatory pathway formed by a series of cholinergic interneurons and final cholinergic motor neurons, apart from the anal contraction.


Chronic hypoxia impairs soluble guanylyl cyclase-mediated pulmonary arterial relaxation in the rat. We have examined the effects of exposing rats to hypoxia (10% fractional inspired O2 concentration) for 2 and 7 days on endothelium-dependent and -independent vasodilation and also on the ability of guanosine 3',5'-cyclic monophosphate (cGMP) to activate cGMP-dependent protein kinase (G-kinase) in rat conduit pulmonary arteries (PA). The ability of acetylcholine (ACh) and sodium nitroprusside (SNP) to both relax PA rings and elevate tissue cGMP levels was significantly attenuated in PA from hypoxic animals. The ability of atrial natriuretic peptide to relax and generate cGMP in PA rings was unchanged by hypoxia. Relaxation and elevation of cGMP levels induced by SNP in aortic rings was unaltered by hypoxia. Similarly, hypoxia did not alter the concentration-dependent activation by exogenous cGMP of G-kinase. We conclude that chronic exposure of rats to hypoxia results in a selective impairment of soluble guanylyl cyclase in rat PA, leading to an attenuation of ACh- and SNP-induced cGMP accumulation and relaxation.


Localization, synthetic regulation, and biology of renal atriopeptin-like prohormone. We recently demonstrated the synthesis and secretion of an atriopeptin (AP)-like prohormone in rat neonatal and adult cortical kidney cell cultures. However, these cultures contained proximal as well as distal tubular epithelial cells; thus characterization of the peptide synthetic cell was not possible. Also, by immunohistochemical techniques, we localized this AP-like prohormone to the distal cortical nephron in adult rat kidney. In this study, we examined further details of the kidney cortical cell type that expresses and secretes this AP-like peptide in adult renal cortical cell cultures, its regulation by adenylate cyclase via adenosine 3',5'-cyclic monophosphate (cAMP) generation, and its ability to stimulate guanylate cyclase. Tubular fragments were derived from cortical tissue of adult Sprague-Dawley rats and separated into four fractions on Percoll density gradient. Cell cultures generated from fraction 3 secreted 5- to 10-fold the amount of this renal peptide compared with fractions 2 and 4. Further cell culture characterization was performed by agonist-stimulated cAMP formation, kallikrein localization, and prostaglandin E2 formation. From these analyses, it was determined that tissue band 3 was enriched for distal cortical connecting tubules. To further evaluate whether mammalian distal nephron synthesizes an AP-like protein, we determined that two immortalized mouse cell lines, derived from either the distal convoluted tubule or cortical collecting tubule, synthesized a radiolabeled AP after being pulsed with [35S]-methionine.(ABSTRACT TRUNCATED AT 250 WORDS)


CGRP and somatostatin modulate chronic hypoxic pulmonary hypertension. Chronic hypoxic pulmonary hypertension (PH), associated with increased pulmonary arterial pressure (PPA) and right ventricular hypertrophy (RVH), correlates significantly with calcitonin gene-related peptide (CGRP) and somatostatin (SOM) levels in lung and blood. CGRP's role in regulation of PPA in chronic hypoxia and its potential interactions with SOM were investigated. CGRP, its antibody (ab) and blocker, CGRP-(8-37), SOM-14, SOM-28, and SOM-ab, respectively, were infused into the pulmonary circulation of hypobaric hypoxia rats for 4, 8, and 16 days. Thereafter, under pentobarbital sodium anesthesia, PPA was measured in the right ventricle and main pulmonary artery. Chronic CGRP infusion prevented PH at all times, whereas immunoneutralization and receptor blocking exacerbated PH. SOM-28 also exacerbated while SOM-14 and SOM-ab decreased PH. RVH generally reflected the PPA. Radioimmunoassay confirmed successful infusion of the peptides with negligible peptide degradation in the pumps throughout 16 days and showed complete immunoneutralization of CGRP with its ab. Peptide levels in lung tissue suggest inhibition of CGRP release by SOM-28 and increased plasma SOM with CGRP infusion. In vitro pharmacological studies suggest that CGRP exerts a receptor-mediated nonadrenergic, nonmuscarinic vasodilatory effect in the lung which is independent of endothelium-derived relaxing factor and does not involve ATP-dependent potassium channels. We conclude that endogenous CGRP plays an important role in pulmonary pressure homeostasis during hypoxia, by directly dilating pulmonary vasculature, thus ameliorating the development of chronic hypoxic pulmonary hypertension in rats.


Contractile actions of C5a on isolated porcine myocardium. Although intracoronary administration of the complement component, C5a, produces deleterious effects on regional coronary blood flow and segmental ventricular function, it is unclear whether a direct myocardial action contributes to the dysfunction induced by the anaphylatoxin. We therefore evaluated the effects of purified porcine C5a on contractile tension of isolated supported ventricular trabeculae from pig hearts. Muscles were studied in a myograph bath at 30 degrees C, electrically stimulated 12 times per minute, and stretched to produce maximal isometric developed tension. C5a concentrations of 30, 100, and 300 ng/ml increased tension (P less than 0.05) 9.8, 5.5, and 20.9%, respectively. In seven of nine muscles exposed to 300 ng/ml C5a, tension initially decreased 10.5% (P less than 0.05) before the positive inotropic effect. Tachyphylaxis was demonstrated by lack of contractile response to a second administration of C5a greater than 70 min after the initial exposure to the complement fragment. Blockade of histamine H1 receptors with diphenhydramine (10(-6) M) markedly attenuated both the positive and negative contractile responses to C5a. beta-Adrenoceptor blockade with propranolol (5.6 x 10(-7) M) did not alter the response to C5a. Levels of thromboxane (Tx)B2, the stable metabolite of TxA2, were augmented in the bath after exposure to C5a (59 +/- 27.3 to 104 +/- 28.2 pg/ml, P less than 0.05). Although the TxA2 agonist, U-46619 (50 ng/ml), significantly increased tension, TxA2 receptor blockade with SQ 29548 (50 ng/ml) did not alter the response to C5a.(ABSTRACT TRUNCATED AT 250 WORDS)


Parasympathetic involvement in rapid meal-associated conditioned insulin secretion in the rat. Blood glucose and plasma insulin concentrations were measured in blood sampled via a cardiac catheter in freely moving rats. To obtain a rapid conditioned cephalic phase of insulin secretion, rats were habituated to one of two feeding schedules. Clock-activated opening of doors in front of the food hopper imposed a feeding schedule of either six meals per day or two meals per day. When the doors were opened in both conditions, insulin increased rapidly during the first minute of feeding in the middle of the light phase. However, when presented an empty food hopper immediately after door opening, only rats in the two meal per day condition showed raised insulin levels and not rats in the six meal per day condition. This response was abolished following pharmacological blockade of nicotinic receptors with hexamethonium and muscarinic receptors with atropine. The present study shows that rapid conditioned insulin secretion can be evoked within one minute by a meal-associated stimulus. These results further indicate that this conditioned insulin secretion is vagally mediated and that its occurrence is dependent on the nature of the feeding schedule.


Ontogeny of DA1 receptor-mediated natriuresis in the rat: in vivo and in vitro correlations. The natriuretic and diuretic effects of dopamine are attenuated in the young. Because dopamine has actions on receptors (e.g., adrenergic, serotonin) other than dopamine, we studied a novel dopamine agonist, pramipexole, which has a selectivity to both DA1 and DA2-receptor subtypes. Intravenous administration of pramipexole resulted in a dose-related (1, 10, and 100 micrograms.kg-1.min-1) increase in urine flow and absolute and fractional sodium excretion and a decrease in mean arterial pressure (MAP) in three groups of rats studied. Pramipexole induced a greater decrease in MAP in 6- to 7- (n = 5) and 9- to 16- (n = 6) than in 3- to 4-wk-old (n = 8) rats; the natriuresis and diuresis were greatest in 12- to 16- and least in 3- to 4-wk-old rats. The renal effects of pramipexole were mainly due to actions at the DA1 receptor, since these effects were completely blocked by the coinfusion of a DA1 antagonist, SKF 83742. To explore further a cause of the attenuated natriuretic effect of pramipexole in the young, we studied the effect of a selective DA1-receptor agonist, fenoldopam, on amiloride-sensitive 22Na+ uptake in renal brush-border membrane vesicles. The 3-s amiloride-sensitive uptake was inhibited (45%) by fenoldopam (5 x 10(-5)M) in 9- to 16- (n = 6) but not in 3- to 4-wk-old (n = 5) rats. These studies suggest that the attenuated natriuretic effect of dopamine in the young is in part due to decreased DA1 action on the brush-border membrane Na(+)-H+ exchanger.


Somnogenic, pyrogenic, and anorectic activities of tumor necrosis factor-alpha and TNF-alpha fragments. Exogenously administered tumor necrosis factor-alpha (TNF-alpha) elicits several symptoms of generalized infections such as fever, increased sleep, and anorexia. The aim of the present work was to localize these effects of TNF-alpha to specific amino acid sequences of the parent molecule by characterizing the in vivo and in vitro activities of several synthetic TNF-alpha fragments. Intracerebroventricular injection of TNF-alpha elicited dose-dependent fevers and increases in non-rapid-eye-movement sleep (NREMS) in rabbits. Four fragments also promoted NREMS and five elicited monophasic fevers. All of the somnogenic fragments share the amino acid sequence 31-36. In rats, TNF-alpha and one of the fragments [TNF-alpha-(69-100)] suppressed 12-h food intake. Furthermore, TNF-alpha increased the expression of the intercellular adhesion molecule-1 and enhanced interferon-gamma-induced HLA-DR expression in human glioblastoma cell line. In contrast, none of the fragments possessed these in vitro activities. Our in vivo results support the concept that there are biologically active regions in the TNF-alpha molecule.


Induction of ICAM-1 by TNF-alpha, IL-1 beta, and LPS in human endothelial cells after downregulation of PKC. The intercellular adhesion molecule 1 (ICAM-1) is induced on endothelial cells by tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and lipopolysaccharide (LPS). We have reported the sensitivity of cytokine-induced ICAM-1 expression to protein kinase inhibitors, including inhibitors of protein kinase C (PKC) [C. L. Myers, S. N. Desai, J. Schembri-King, G. L. Letts, and R. W. Wallace. Am. J. Physiol. 262 (Cell Physiol. 31): C365-C373, 1992]. To directly investigate the role of PKC in ICAM-1 induction, we downregulated PKC by pretreatment of human umbilical vein endothelial cells with phorbol 12-myristate 13-acetate (PMA) and assessed ICAM-1 protein and mRNA induction elicited by subsequent exposure to inflammatory stimuli. PMA treatment results in ICAM-1 protein induction that declines to basal levels by 3 days. Western blots of endothelial cell lysates reveal a nearly complete loss of immunologically reactive PKC. Subsequent activation with cytokine or LPS leads to reinduction of ICAM-1 protein and mRNA; however, the cells no longer produced substantial amounts of ICAM-1 protein or mRNA in response to PMA stimulation. Cross desensitization is observed with phorbol dibutyrate, while 4 alpha-phorbol has no desensitizing effect. The data indicate that PKC activation, while capable of inducing ICAM-1 expression, is not essential for ICAM-1 induction by the inflammatory mediators TNF-alpha, IL-1 beta, or LPS.


Expression of aminopeptidase N in fetal rat lung during development. The ectopeptidase, aminopeptidase N, serves as a cell surface marker of the apical surface of the alveolar type II epithelial cell in adult lung. It is also present in fetal lung before differentiation of morphologically mature type II alveolar epithelial cells, suggesting that it is expressed by precursors of the type II cells. We have examined the mRNA coding for the aminopeptidase in adult and fetal lung and in mature type II cells and determined levels of mRNA and immunoreactive protein during fetal lung development. Comparison of the temporal patterns of steady-state levels of aminopeptidase mRNA and immunoreactive protein during development show that the expression of the protein is developmentally regulated and that expression is regulated, at least in part, at a pretranslational level. Both mRNA and immunoreactive protein levels increase severalfold on the final gestational day, suggesting that the function of the aminopeptidase may be associated with air breathing.


Cyclosporin and quinidine inhibition of renal digoxin excretion: evidence for luminal secretion of digoxin. We studied the in vivo luminal and contraluminal uptake of [3H]digoxin in dog kidney using the single-pass multiple indicator dilution method. A bolus tracer of 125I-albumin (plasma reference), creatinine, or L-[14C]glucose [extracellular reference (ecf)] and [3H]digoxin (or [3H]ouabain) was injected into the left renal artery, and timed serial samples were collected from the left renal vein (basolateral uptake) and left and right ureters (luminal uptake). [3H]ouabain was excreted solely by filtration and exhibited saturable and irreversible binding at the basolateral surface. Uptake of [3H]digoxin across the basolateral membrane was large and nonsaturable. Despite urine flow-dependent reabsorption and approximately 20% protein binding, the urine recovery ratio for [3H]-digoxin/glomerular (ecf) marker was 0.97 +/- 0.04 (n = 29), indicating net digoxin secretion. After intravenous infusions of cyclosporin in Cremophor EL (0.5-3.5 microM), the urine recovery ratio decreased in a dose-dependent manner from control values of 1.13 +/- 0.06 (n = 12) to 0.62 +/- 0.03 (n = 14). There was no change in the relative renal vein recovery. Left renal artery infusion of quinidine (37.5 micrograms.min-1.kg-1) decreased the relative urine recovery of [3H]digoxin by 46% (n = 6) but had no effect on postglomerular extraction. Cyclosporin and quinidine are known inhibitors of P-glycoprotein. But digoxin did not compete with [3H]azidopine for binding in rat brush-border membranes or membranes prepared from the multidrug-resistant cell line CHRC5. The exact mechanism for renal digoxin secretion remains to be determined, but our results point to a luminal localization of this secretory system.


Age-dependent changes in alpha-adrenoceptor-mediated contractility of isolated human resistance arteries. Human subcutaneous resistance arteries (122-298 microns), isolated from 139 patients undergoing surgery, were mounted in an isometric myograph. With the use of multiple regression analysis, five different modes of activation were examined for possible associations with age and mean arterial blood pressure of the patients: the contractile responses to 10 microM norepinephrine (mixed alpha 1-agonist/alpha 2-agonist), perivascular nerve stimulation, 10 microM phenylephrine (alpha 1-agonist), 100 microM B-HT 933 (alpha 2-agonist), and depolarization by potassium chloride. Blood pressure increased significantly with age. Blood pressure independently was not correlated to any mode of activation. With increasing patient age, however, responses to norepinephrine, phenylephrine, and perivascular nerve stimulation decreased, whereas the response to B-HT 933 increased; responses to potassium chloride were unaltered. Also corrected for changes in blood pressure, age independently was negatively correlated to the response to norepinephrine and phenylephrine, whereas a positive, though nonsignificant (P value = 0.12), correlation was observed between age independently and the response to B-HT 933. These data suggest that the ability of isolated human resistance arteries to evoke contractions medicated by postjunctional alpha 1-, but not alpha 2-adrenoceptors, decreases with age.


Ontogeny of renal response to specific dopamine DA1-receptor stimulation in sheep. The present study was designed to characterize the developmental changes in the renal responses to dopamine DA1-receptor activation in chronically instrumented preterm (109-115 days) and near-term (130-140 days, full term 145 days) fetal sheep. Cumulative doses of the selective DA1-agonist fenoldopam increased mean arterial blood pressure (MABP) in both preterm (+16 +/- 3%) and near-term fetuses (+16 +/- 3%) but had no significant effect on renal blood flow velocity. Infusion of the DA1-antagonist SCH-23390 did not affect the increase in MABP, suggesting that the effect of fenoldopam on MABP was not directly related to activation of DA1-receptors. Fenoldopam infusion had no significant effects on renal function parameters in preterm fetuses. In near-term fetuses, however, fenoldopam increased urinary flow rate (82.6 +/- 20.9%, P < 0.003), glomerular filtration rate (GFR; 16.6 +/- 4.9%, P < 0.01), urinary sodium excretion (40.1 +/- 14.9%, P < 0.02), and fractional excretion of sodium (26.8 +/- 11.2%, P < 0.03). Infusion of the DA1-antagonist SCH-23390 blocked the fenoldopam-induced diuresis and natriuresis but had no significant effect on the rise in GFR. Fenoldopam infusion had no significant effects on plasma renin activity and plasma aldosterone concentration and on urinary prostaglandin (PG) excretion (PGE2, PGF2 alpha, and 6-keto-PGF1 alpha). Taken together, these results suggest that the renal effect of DA1-receptor activation is age dependent and that stimulation of DA1-receptor in near-term fetuses is associated with a diuresis and natriuresis that seem to be independent of renal hemodynamics and adrenal effects.


Potentiation of thermoregulatory responses to isoproterenol by beta-adrenergic antagonists. The thermoregulatory effects of isothermogenic doses of isoproterenol (Iso) and a novel beta-agonist (BRL 35135) were tested in rats at 22 degrees C and in rats trained to bar press for radiant heat at -8 degrees C. BRL 35135 produced hyperthermia at 22 degrees C and reduced operant responding for heat at -8 degrees C, whereas Iso reduced body temperature and increased operant responding. In both situations, the negative effects of Iso on thermal balance were abolished by propranolol at doses that did not inhibit heat production. In anesthetized rats, propranolol potentiated the Iso-induced rise in brown adipose tissue and colonic temperature. The potentiation was more marked with the beta 2-selective antagonist ICI 118,551, whereas treatment with the beta 1-selective antagonist atenolol resulted in a profound Iso-induced reduction in temperature. The two selective antagonists also produced divergent responses in operant behavior in Iso-treated rats at -8 degrees C. These experiments demonstrate the extent to which responses to a nonselective agonist can be manipulated using appropriately low doses of selective antagonists and indicate that the effects of Iso on thermal balance are due to its beta 2 activity.


Impact of psychiatric hospitalization on behavioral complications of Alzheimer's disease. The authors conducted a prospective study of the clinical utility of the four DSM-III-R subtypes of primary degenerative dementia of the Alzheimer type (with delirium, with delusions, with depression, or uncomplicated) and acute psychiatric hospitalization for treatment of these subtypes. The subjects were 120 consecutive inpatients with Alzheimer's disease, most of whom had behavioral abnormalities. Each subject received detailed physical, neurological, psychiatric, and mental status examinations. The presence or absence of specific behavioral problems was also documented. Patients were treated with medication, psychotherapy, and behavioral techniques. While all patients could be assigned to one of the four DSM-III-R behavioral subtypes, the uncomplicated subtype did not accurately reflect the burden of behavioral symptoms in the patients who did not have delirium, delusions, or depression. Each behavioral subtype responded in a characteristic way to inpatient treatment, as reflected by changes in scores on four psychometric scales used to assess cognitive impairment, psychiatric symptoms severity, and level of functioning at admission and at discharge, as well as by changes in residential setting following hospitalization. Half of all patients admitted from their homes and two-thirds of those with depression were able to go home following discharge. Behavioral syndromes in Alzheimer's disease should not be overlooked, because they have both clinical and prognostic significance. Short-term psychiatric hospitalization is effective and efficient for achieving the goal of returning patients to their homes and for safely implementing specific treatments in this frail population, and it may reduce the need for institutionalization.


A national study of psychiatrists' professional activities. A mail survey was conducted in 1988-1989 to study the professional activities of U.S. psychiatrists. Data from the 19,431 active respondents are reported. Nineteen percent of the psychiatrists were women, an increase from the 17% reported in 1982. The median age of the respondents was 50 years. Nearly one-third of the respondents expressed interest in each of the following areas of subspecialization: adolescent psychiatry, substance abuse, geriatrics, and consultation-liaison psychiatry. More than one-fifth reported formal fellowship training in child/adolescent psychiatry. The psychiatrists worked an average of 48 hours per week--two-thirds in direct patient care--in an average of 2.3 different settings. The proportion of psychiatrists reporting private practice as their primary work setting showed a marked decline from 53% in 1982 to 45% in 1988. There was an increase from 4% in 1982 to 11% in 1988 in those whose primary work setting was a private psychiatric hospital. The typical caseload was over 60 patients, with roughly half that number seen each week. For inpatients treated, the two most common diagnoses were affective disorders and schizophrenic disorders. In a typical week psychiatrists treated about one-half of their outpatients with individual psychotherapy; three-fifths of these were also treated with medications. The average net income for psychiatrists working 35 hours or more per week was $99,850 for men and $73,174 for women. Major trends evident from this study are subspecialization, medicalization, privatization, feminization, and organizational diversification.


Liver adenomatosis with granulomas in two patients on long-term oral contraceptives. We report two cases of liver cell adenomatosis associated with granulomatous hepatitis, both developed in white women (52 and 39 years of age) on long-term oral contraceptives (for 18 and 12 years, respectively). The first patient underwent surgery for five hepatic tumors 1-7 cm in diameter; the other patient had a partial segmentectomy for a 4-cm hepatic nodule of the right lobe. In both cases, dissection of the liver showed many other diffuse and smaller nodules. Histologically, all tumors were benign liver cell adenomas, with cellular atypia in the largest tumor and associated in both cases with granulomatous hepatitis, with numerous noncaseating epithelioid and giant cell granulomas located either only in the tumors (case 1) or diffusely in the tumoral and nontumoral hepatic parenchyma (case 2). During follow-up, ultrasound showed new nodular lesions in case 2, whereas in case 1 evolution was uneventful. In estroprogestative-associated liver diseases, adenomas are common, but adenomatosis and granulomas are rare. An association of these latter two conditions would seem to be exceptional. The prognosis for adenomatosis remains uncertain.


Practicality of molecular studies to evaluate small lymphocytic proliferations in endoscopic gastric biopsies. Fifteen endoscopic gastric biopsies (GBx) from 12 patients with small lymphocytic infiltrates morphologically raising a differential of indeterminate lymphocytic infiltrate versus mucosa-associated lymphoid tissue (MALT) lymphoma were analyzed genotypically after frozen-section identification of the abnormal lymphocytic infiltrate. Frozen-section immunoperoxidase immunophenotyping was equivocal in each case. All patients had abdominal pain attributable to superficial gastric ulceration, most often antral, without peripheral lymphadenopathy or hepatosplenomegaly. Rearrangement of the immunoglobulin heavy-chain gene (JH-R, seven patients) or kappa light-chain gene (JK-R, eight patients), was found in eight GBx from eight (seven stage IAE; one stage IBE) of 12 patients, establishing, in conjunction with the histologic features, a diagnosis of low-grade B-cell lymphoma. This diagnosis had not been tenable on multiple prior GBx, ranging from one to five per patient, over intervals of 1 month to 6.5 years (median 4.5 months). The T-cell receptor beta-chain gene retained germline configuration in all cases. Insufficient DNA for molecular studies was extracted from the GBx of two patients, one with JK-R (JH-G) on subsequent GBx and one without further GBx. One patient had two GBx, each demonstrating a single additional band in HindIII digests hybridized with the JH probe. No rearrangements were detected in either the BamHI or the EcoRI digests. Uninvolved tissue from this patient was not available for the exclusion of restriction fragment length polymorphism. Three GBx (two patients) showed germline JH genes (JH-G). One had a partial gastrectomy (histology: MALT lymphoma) in 1981 followed by GBx in 1983 (histologically benign) and in 1990 (JH-G), and negative esophagogastroduodenoscopy (EGD) in 1991 without biopsy. The other patient (two GBx with JH-G) had multiple subsequent abnormal EGD, but no biopsies since December 18, 1990. Adequate DNA for gene rearrangement studies can be extracted from GBx samples weighing as little as 20 mg. The two samples with insufficient DNA weighed 1 and 16 mg, respectively. Practically speaking, the remainder of a frozen block from a single GBx is adequate, thus allowing the screening of multiple endoscopic GBx by sequential frozen sections to determine which one contains the most extensive lymphocytic infiltrate for molecular study. Consistent results are obtained on samples weighing 40 to 60 mg. This method is a suitable alternative to kappa/lambda frozen-section immunoperoxidase immunostaining, which can be uninterpretable on endoscopic biopsies or small biopsies from other sites.


Prognostic significance of co-expression of c-erbB-2 oncoprotein and epidermal growth factor receptor in breast cancer patients. The expression of c-erbB-2 oncoprotein and epidermal growth factor receptor (EGFR) was examined by immunocytochemical and radioreceptor assays in 115 patients with primary breast cancer. In 48 of 115 patients (42%), the assays were found to be positive for the expression of c-erbB-2 oncoprotein, and, in 44 of 115 (35%) patients, the assays were positive for the expression of EGFR. There was no correlation between the expression of c-erbB-2 oncoprotein and EGFR. Clinical survey demonstrated that both c-erbB-2 oncoprotein expression and EGFR expression have independent prognostic values. Furthermore, when patients were divided into three groups on the basis of the expression of both c-erbB-2 oncoprotein and EGFR, those who were found to be positive for the expression of both c-erbB-2 oncoprotein and EGFR showed a worse prognosis than other groups. These results suggest that the combination of the expression of both c-erbB-2 oncoprotein and EGFR may be important in selecting patients who have a poor prognosis.


Stress fractures in ballet dancers.

Stress fractures in ballet dancers. We surveyed 54 female dancers in two professional ballet companies. A total of 27 fractures were reported in 17 dancers. Metatarsal fractures were the most common (63%), followed by fractures of the tibia (22%) and spine (7%). Dancers who danced greater than 5 hours per day were significantly more likely to have a stress fracture than those dancing less than 5 hours per day. Dancers in the stress fracture group also had a significantly longer duration of amenorrhea than those in the group with no stress fractures. No significant difference was found between the dancers who had stress fractures and those who did not with regard to any of the other variables examined. These data suggest that prolonged amenorrheic intervals and heavy training schedules may predispose ballet dancers to stress fractures. Of the 17 dancers with stress fractures, only 1 had neither of these risk factors.


Treatment of adenocarcinoma of the pancreas with somatostatin and gonadoliberin (luteinizing hormone-releasing hormone). The French Associations for Surgical Research. Experimental studies have shown a significant inhibition of adenocarcinoma of the pancreas by gonadoliberin (luteinizing hormone-releasing hormone [LH-RH]) and somatostatin. The aim of this prospective randomized study was to compare the potential value of somatostatin (250 micrograms every 8 hours), LH-RH (3.75 mg monthly), or combined, to a control group. One hundred sixty-three patients with adenocarcinoma of the pancreas who did not undergo resection for cure were divided into 4 groups that did not differ in terms of clinical, biologic, or pathologic data. The mean survival times were 6 months in the LH-RH plus somatostatin group, 5.5 months in the LH-RH group, 4.3 months in the control group, and 3.8 months in the somatostatin group. However, the life-table analyses for all randomized patients, and separately according to sex, the lymph node extension, and metastatic spread were not different between groups. Improvement of patient status was observed in 20% of the patients receiving hormone therapy without any difference noted between the treatment regimens. These disappointing results may be explained by the degree of extension of pancreatic carcinoma in the patients studied. The results suggest that different hormone therapy regimens might be considered according to the age and the sex of patients, as well as to the presence or absence of hormone receptors.


Vecuronium: an anthropometric comparison. This study set out to determine if there was any resistance to vecuronium in Nepalese studied in Nepal compared with Nepalese, Chinese and European patients studied in Hong Kong. The four groups, each of 10 male and 10 female patients, were intubated 60 s after administration of 0.1 mg.kg-1 vecuronium. The Nepalese patients in Nepal had significantly less satisfactory intubating conditions (p = 0.002). Similarly, male patients had significantly less satisfactory conditions than female patients (p = 0.004). Some anthropometric measurements were significantly different between the patients in Nepal and those in Hong Kong. There were also sex-related anthropometric differences. It is suggested that differences in response to vecuronium could be explained by differences in distribution volume and muscle mass.


Characterization of glucose microsensors for intracellular measurements. Ultrasmall glucose sensors have been constructed by using platinum-deposited carbon ring microelectrodes with glucose oxidase. Response times as low as 270 ms have been obtained with these sensors. Moreover, there is a linear relationship between sensor tip diameter and response times. The use of these sensors has been demonstrated in the detection of glucose in single-cell cytoplasm of the large dopamine cell of the pond snail Planorbis corneus. Current responses obtained at these sensors implanted into a cell increase following injection of 2 pL of glucose solution (3 M) into the cell. Results obtained from these experiments show that these sensors are suitable for glucose monitoring in ultrasmall environments. In addition, characterizations of these sensors have been investigated under different O2 concentrations. At atmospheric oxygen concentrations, glucose levels in the submillimolar range can be measured without oxygen interference; however, oxygen interference can be substantial at low oxygen concentrations.


Efficacy of anticholinergic and beta-adrenergic agonist treatment of maximal cholinergic bronchospasm in tracheally intubated rabbits. Cholinergically induced bronchoconstriction is thought to be a major cause of bronchospasm during anesthesia. We used tracheally intubated rabbits (4-mm endotracheal tube) stimulated with methacholine to assess the efficacy of beta-adrenergic agonist and anticholinergic treatment in reversing the increases in respiratory system resistance. Four groups were compared: (a) inhaled metaproterenol, 20 puffs via metered dose inhaler (0.65 mg/puff); (b) inhaled ipratropium bromide, 20 puffs from a metered dose inhaler (18 micrograms/puff); (c) 2 mg of intravenous atropine; and (d) no treatment after methacholine challenge as a control group. Methacholine increased respiratory system resistance from 0.041 +/- 0.001 (mean +/- SEM) to 0.098 +/- 0.006 cm H2O.mL-1.s-1 (P < 0.001). Whereas beta-adrenergic agonist treatment was ineffective in ameliorating bronchoconstriction, inhaled ipratropium bromide and atropine were highly effective, causing an 86%-88% reversal in the methacholine-induced increase in respiratory system resistance. Both these agents were also effective in improving dynamic compliance. We conclude that inhaled ipratropium bromide is effective in treating cholinergic bronchospasm even when administered via a small endotracheal tube and that the beta-adrenergic agonist metaproterenol is ineffective in rabbits in the face of maximal cholinergic stimulation.


Total intravenous anesthesia: effects of opioid versus hypnotic supplementation on autonomic responses and recovery. During radical prostatectomy procedures under total intravenous anesthesia, acute hemodynamic responses to retropubic dissection (30% +/- 8% to 36% +/- 12% [mean +/- SD] increases in mean arterial pressure) were treated with supplemental doses of either an opioid analgesic (alfentanil) or a sedative-hypnotic (propofol) to return the mean arterial pressure to within 10% of the preincision value. Although both drugs were effective, control with propofol required 10.1 +/- 2.5 min compared with 6.3 +/- 2.6 min in the alfentanil group (mean +/- SD; P < 0.01). Plasma stress hormone concentrations increased significantly in response to this surgical stimulus: epinephrine increased from 246% +/- 169% to 283% +/- 330%; norepinephrine increased from 44% +/- 33% to 83% +/- 104%; and antidiuretic hormone increased from 1300% +/- 1591% to 1700% +/- 1328%. Both alfentanil and propofol were equally effective in returning the catecholamine concentrations to their preincision levels. However, antidiuretic hormone levels remained above preincision values in both groups. Despite earlier awakening (3.4 +/- 2.9 vs 9.1 +/- 6.8 min; P < 0.05) in the alfentanil treatment group, there was no difference in time to spontaneous ventilation or tracheal extubation between the groups. In addition, 36% of the alfentanil-treated patients required antihypertensive therapy in the postanesthesia care unit compared with only 9% in the propofol group (P < 0.05). Postanesthesia care unit and hospital discharge times were similar in both treatment groups. We conclude that supplemental doses of alfentanil or propofol were equally effective in controlling acute hemodynamic and hormonal responses to surgical stimuli during total intravenous anesthesia.


Defining the dose range for esmolol used in electroconvulsive therapy hemodynamic attenuation. We evaluated the clinical effectiveness of esmolol, an ultra-short-acting, beta-adrenergic receptor blocking drug, to control the sinus tachycardia and increase in arterial blood pressure induced by electroconvulsive therapy (ECT). Each of 20 patients, ASA physical status I-III, participated in a double-blind, randomized Latin-Square study involving two matched-pair trials (placebo versus esmolol given as a 500-micrograms/kg bolus followed by either 300 micrograms.kg-1.min-1 [high dose], 200 micrograms.kg-1.min-1 [medium dose], or 100 micrograms.kg-1.min-1 [low dose] infusion of esmolol) during ECT. Each patient acted as his or her own control (total number of ECT procedures were 160). We administered a 1-min bolus of placebo (normal saline) or esmolol at the rate of 500 micrograms.kg-1.min-1 followed by either high-, medium-, or low-dose esmolol or placebo for an additional 3 min. We then induced anesthesia with methohexital (1 mg/kg) and succinylcholine (0.5 mg/kg) IV. Ninety seconds after the administration of succinylcholine, the electrical stimulus was applied to induce seizure. The infusion of placebo or esmolol was discontinued 3 min after the electrical stimulus. Significant decreases were found in mean heart rate from minute 3 until minute 7 and in the maximum heart rate. The mean of each patient's maximum heart rate after seizure changed from 147 +/- 18 bpm in placebo patients to 112 +/- 20 bpm in high-dose esmolol patients; to 121 +/- 23 bpm in medium-dose esmolol patients; and to 124 +/- 20 bpm in low-dose esmolol patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Tetanus-induced changes in apparent recovery after bolus doses of atracurium or vecuronium. The current study evaluated the duration and magnitude of post-tetanic effects in 56 patients recovering from a bolus dose of nondepolarizing relaxant to assess the impact of tetanus on monitoring in a common clinical setting. After induction of general anesthesia (thiopental, fentanyl, oxygen, nitrous oxide, and isoflurane), a baseline response to train-of-four (TOF) stimulation was recorded using an adductor pollicis force transducer, and the ratio of the fourth response (T4) to the first (T1) was calculated. Patients then received a bolus dose of either atracurium 0.50 mg.kg-1 (n = 28) or vecuronium 0.10 mg.kg-1 (n = 28). TOF was recorded at 12-s intervals between 25% and 75% recovery of T1 (time25-75%, first data set); then, block was reestablished with the same agent (atracurium 0.10 or vecuronium 0.02 mg.kg-1), and monitoring of time25-75% was repeated (second data set). Subjects were randomized such that none, one, or both sets had TOF monitoring interrupted by a 5-s, 50-Hz tetanic stimulus at 50% recovery (TET). For each drug, 7 patients were assigned to each of the four possible sequences: no tetanus (NOTET) set followed by NOTET set; NOTET-TET; TET-NOTET; and TET-TET. After either drug, the TET data sets demonstrated significant acceleration of recovery of T1 from 50% to 75% (time50-75%) of its baseline height (P less than 0.05 by paired t test). After atracurium, time50-75% was shortened by the tetanic stimulation from a control of 6.3 +/- 1.1 to 5.0 +/- 1.3 min (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


AMPA and NMDA receptor antagonists do not decrease hippocampal glutamate concentrations during transient global ischemia. Increased extracellular concentrations of glutamate during episodes of cerebral ischemia may be due in part to a positive glutaminergic feedback loop. We evaluated the effect of selective AMPA or NMDA receptor antagonists on hippocampal extracellular concentrations of excitatory amino acids during ischemia and reperfusion. Thirteen New Zealand white rabbits were subjected to 10 min of global cerebral ischemia produced by neck tourniquet inflation (20 psi) combined with systemic hypotension during halothane (1-1.5%) anesthesia. Hippocampal extracellular concentrations of glutamate, aspartate, and glycine were monitored using in vivo microdialysis. NBQX (a selective AMPA receptor antagonist), MK801 (a noncompetitive NMDA receptor antagonist), or 5% dextrose was administered starting 1 h before ischemia. The NBQX group (n = 4) received 5 mg.kg-1 of NBQX intravenously (dissolved in 5% dextrose) over 5 min followed by an infusion of 5 mg.kg-1.h-1. The 5% dextrose group (n = 4) received an equivalent volume of 5% dextrose. The peak concentrations of glutamate, aspartate, and glycine in the early reperfusion period were 5-8-fold, 9-10-fold, and 4-5-fold higher than preischemic values, respectively. There were no significant differences, however, among the three groups in the concentrations of glutamate, aspartate, or glycine at any time during the study. These results do not support the existence of a positive feedback loop for glutamate mediated via AMPA or NMDA autoreceptors in the hippocampus during transient global ischemia or reperfusion.


Sheep linkage mapping: restriction fragment length polymorphism detection with heterologous cDNA probes. A selection of cattle, human and sheep cDNA probes were screened against sheep genomic DNA, cut with 10 different restriction enzymes, to assess the usefulness of these probes for restriction fragment length polymorphism (RFLP) linkage studies in sheep. Two-thirds of the cattle cDNA probes showed moderate to strong homology with sheep DNA samples, compared with less than half of the human cDNA probes at the final washing stringency chosen for the experiments. The set of probes tested detected a useful frequency of RFLPs. Fifty-seven per cent of probes showing moderate to strong homology identified RFLPs with one or more restriction enzymes. Restriction enzymes that detected RFLPs most frequently in sheep were TaqI and MspI. The results show that sheep and cattle cDNA probes, including candidate genes for production traits, identified a high frequency of RFLPs suitable for genetic mapping in sheep.


Detection of bovine alpha s1-casein genomic variants using the allele-specific polymerase chain reaction. A method was developed to distinguish between genotypic variants B and C of bovine alpha s1-casein, using the allele-specific polymerase chain reaction (ASPCR). The alpha s1-casein genotype determined for 17 Jersey cows by the ASPCR method was confirmed by typing the alpha s1-casein milk proteins on isoelectric focusing gels. Using the ASPCR method described, rapid analysis of the alpha s1-casein genotype of bulls is now possible. In addition, kappa-casein genotypes can be determined from the same PCR reaction.


[The prevention of undesirable changes in the endocrine system in patients in the intraoperative period using stress-preventive preparations]. The functions of the adrenal cortex, thyroid gland and carbohydrate metabolism have been analysed in 20 patients to whom current schemes of premedication and anesthesia have been applied, 20 patients who in the nearest preoperative period and intraoperatively have been administered ganglioblockers and adrenolytics, and 20 patients in whom clophelin has been used during premedication and anesthesia. It has been established that the use of stress-protecting agents for premedication and anesthesia almost prevents hyperergic reaction of the adrenal cortex, thyroid gland and carbohydrate metabolism.


Biological properties and genetic analysis of the ompA locus in chlamydiae isolated from swine. Eight strains of Chlamydia psittaci isolated from swine with pneumonia, pleuritis, pericarditis, and enteritis were characterized through analysis of the major outer membrane protein gene ompA by a two-step polymerase chain reaction, by their interactions with cells in culture, and by the morphologic features and ultrastructure of intracellular inclusions. Amplified chlamydial ompA DNA fragments were differentiated by restriction endonuclease digestion. Chlamydial isolates were separated into 2 types on the basis of ompA restriction fragment length polymorphism. Strains of type L71 had finely granular inclusions, whereas those of type 1710S contained pleomorphic reticulate bodies (RB) in the inclusions, which are characteristic of aberrant chlamydial developmental forms. Chlamydial types L71 and 1710S required centrifuge-assisted inoculation for efficient infection of cell cultures. Cultivation in cell culture medium containing cycloheximide increased the numbers of chlamydial inclusions about 1.5-fold. These strains formed few elementary bodies in yolk sac cells of chicken embryos. Ultrastructurally, unique doublet RB were observed, particularly in strains of the ompA type L71. These doublets consisted of 2 RB, bounded by a cytoplasmic membrane, contained within a common cell wall and an extended periplasmic space. Ultrastructural examination of strains of the ompA type 1710S confirmed the aberrant chlamydial developmental forms, but evidence of viral infection of the RB as a cause of these aberrant forms was not found. The strain S45 isolated from intestinal sites of swine was a trachoma restriction fragment length polymorphism type. With the mouse biotype, it represented the second isolate from animals of Chlamydia trachomatis.


Pathogenicity of porcine enterotoxigenic Escherichia coli that do not express K88, K99, F41, or 987P adhesins. Three-week-old weaned and colostrum-deprived neonatal (less than 1 day old) pigs were inoculated to determine the pathogenicity of 2 enterotoxigenic Escherichia coli isolates that do not express K88, K99, F41, or 987P adhesins (strains 2134 and 2171). Strains 2134 and 2171 were isolated from pigs that had diarrhea after weaning attributable to enterotoxigenic E coli infection. We found that both strains of E coli adhered in the ileum and caused diarrhea in pigs of both age groups. In control experiments, adherent bacteria were not seen in the ileum of pigs less than 1 day old or 3 weeks old that were noninoculated or inoculated with a nonpathogenic strain of E coli. These control pigs did not develop diarrhea. Antisera raised against strains 2134 and 2171 and absorbed with the autologous strain, grown at 18 C, were used for bacterial-agglutination and colony-immunoblot assays. Both absorbed antisera reacted with strains 2134 and 2171, but not with strains that express K99, F41, or 987P adhesins. A cross-reaction was observed with 2 wild-type K88 strains, but not with a K12 strain that expresses K88 pili. Indirect immunofluorescence with these absorbed antisera revealed adherent bacteria in frozen sections of ileum from pigs infected with either strain. We concluded that these strains are pathogenic and express a common surface antigen that may be a novel adhesin in E coli strains that cause diarrhea in weaned pigs.