Interleukin-2 deficit in hemodialysis patients. Role of prostaglandins. Uremic patients suffer from various immunological alterations, whose pathogenesis is still unknown. Here, we studied 37 hemodialysis patients in order to investigate the role of prostaglandins (PGs) in uremic immunological deficiency, specifically in relation to interleukin-2 (IL-2) synthesis. We confirmed previous published data on deficient response to PHA in chronic renal failure patients (cpm, mean +/- SEM: 15,400 +/- 2,100 in uremics vs. 29,500 +/- 3,380 in controls, p < 0.04) and established a correlation between this deficiency and diminished IL-2 synthesis (r = 0.619, p < 0.05). The direct measurement of PGs in lymphocyte cultures showed greatly increased concentrations in the presence of uremic serum (US). We found that PGs synthesis can be inhibited by up to 80% if cultures are supplemented with indomethacin (IND--a cyclooxigenase inhibitor) or by removal of monocytes (producers of PGs). Both methods situated the uremic proliferative response within the normal range in cultures with FCS, and close to the normal range in cultures with US. We observed a deficit of IL-2 in hemodialysis patients (means +/- SD: 8,940 +/- 6,420 in uremics vs. 16,900 +/- 3,890 in controls). Addition of exogenous IL-2 normalized lymphocyte response even in US cultures, with no additive effect between PGs inhibition and exogenous IL-2 except in US cultures. It is suggested that IL-2 deficit of uremics depends, at least in part, on an increase in PGs synthesis induced by US.(ABSTRACT TRUNCATED AT 250 WORDS)


Performance characteristics of contemporary hemodialysis and venovenous hemofiltration in acute renal failure. Modality choice in the treatment of acute renal failure (ARF) should be based on the match between individual patient needs and the characteristics of available therapies. Considerations include access, risk of bleeding, hemodynamic instability, and ability to remove excess volume. We prospectively studied 547 consecutive treatments for ARF in 110 patients to determine the performance features of modalities based on single-vessel venous access: hemodialysis (HD) and venovenous hemofiltration (VVH). All treatments were performed in an 18-month period at a single center. Patients' ages ranged from 16 to 84 years; 26 were trauma cases; 69 patients expired during their hospitalization. Mean number of treatments per patient was 4.97. Mean treatment duration was 197 min. Heparin was used unless active bleeding was present or felt to be a high risk; mean dose required was 2628 units. Pressor therapy was in progress prior to initiation of 260 (48%) of treatments. pretreatment and posttreatment mean systolic and diastolic BP were unchanged for the group. Forty-three (7.9%) treatments were terminated prematurely; of these only 27 (4.9%) were due to hypotension. No patient developed clinically apparent bleeding during any treatment. Contemporary equipment and techniques allow for provision of high-quality intermittent therapy for ARF, with excellent hemodynamic stability. Shorter, single-vein access treatments are advantageous for severely ill or injured patients who often undergo invasive monitoring and multiple studies or procedures.


Influenza vaccination in chronic hemodialysis patients. The effect of zinc supplementation. Since influenza increases the mortality of chronically ill patients we decided to study the effectiveness of influenza vaccination in hemodialysis (HD) patients. Nineteen HD patients aged from 20 to 60 years, on unrestricted diet and with no febrile episode, were studied. Blood samples were collected before the intramuscular injection of 0.5 mL multivalent influenza vaccine (Inflexal Berna) and every 2 weeks thereafter. At the end of 4th week a second vaccination was done and a dosage of 200 mg of zinc acetate (60 mg elemental zinc) was given daily to each patient for at least 4 weeks. Before vaccination the antibody titers to influenza virus ranged from 1:10 to 1:80 and after vaccination from 1:20 to 1:640. Four weeks after vaccination 6/19 (31.5%), 8/19 (42%), and 10/19 (52.5%) patients showed a fourfold or greater increase at serum antibody titers to antigens A/Singapore, A/Sichuan, and B/Beijing, respectively. The zinc supplementation after the second vaccination induced a similar increase of serum antibody titers to the A/Singapore but some even greater increase of the antibody titers to the A/Sichuan and B/Beijing. Serum immunoglobulins and complement components C3/C4 were not changed during this study. It is suggested that about 50% of uremic patients respond to the influenza vaccination and that zinc treatment does not increase this responsiveness.


Evaluation and first validation study on a simplified drug dosage algorithm for multiple organ failure patients. As reported previously, drug concentrations during continuous hemofiltration (HF) and extracorporeal lung assist (ELA) follow certain rules, which can be expressed by a simplified algorithm for dosage adjustment: Drug sieving (S, fu) depends on the protein free fraction with small limitations, while the extrarenal elimination rate is not a constant but correlates inversely with the clinical state, r = -0.34, p = 0.00067, n = 96. Up to now, more than 218 cases of drug dosage adjustments have been performed, following the described regimen: The expected concentration is obtained in 79-84% already from the first estimation for drugs such as aminoglycosides, vancomycin, teicoplanin, beta-lactam antibiotics, heart glycosides, and theophylline. Skilled therapeutic drug monitoring (TDM) with elaborated pharmacokinetic programs fails to improve these results significantly. Nevertheless, sporadic TDM is essential in these patients according to their rapidly changing clinical states.


Bardet-Biedl syndrome and cystinuria.

Bardet-Biedl syndrome and cystinuria. An unusual association of Bardet-Biedl syndrome with cystinuria was described in one patient. A 21-year-old male was admitted to hospital because of renal failure, severe deterioration of visual acuity, polydactyly, brachydactyly, and mental retardation. Laboratory investigations revealed a serum creatinine of 292 mumol/L (3.3 mg/dL) and a GFR of 25 mL/min per 1.73 m2. Quantitative ion exchange chromatography demonstrated an increased urinary excretion rate of cystine, lysine, arginine, and ornithine. The ophthalmologic examination showed a severe atypical retinal dystrophy. Visual acuity was severely deteriorated and the patient could only count the examining physician's fingers. The patient had been previously evaluated at the age of 7 years for polyuria, polydipsia, and growth failure. His workup at that time demonstrated nephrogenic diabetes insipidus, normal GFR, and a urinary amino acid pattern consistent with the cystinuric phenotype. There was mental retardation notwithstanding the normal ophthalmologic examination. Intravenous pyelography showed calyceal clubbing, calyceal cysts, and lobulated renal outlines of the fetal type. The patient was evaluated again at the age of 13 years for deterioration of visual acuity and the ophthalmologic examination showed an atypical retinal dystrophy, with sparse pigmentation, central and peripheral atrophy, attenuated vessels, and marked optic disk pallor. To our knowledge the association of Bardet-Biedl syndrome with cystinuria has never been reported. It is unlikely that cystinuria may have contributed to the kidney damage. The possibility that mental retardation has been induced or aggravated by cystinuria cannot be excluded.


Acute renal failure in severe hypothermia. A patient who developed acute renal failure associated with severe hypothermia is reported. Warm peritoneal dialysis was initiated for core rewarming followed by intermittent hemodialysis till he entered the diuretic phase. The factors which led to acute renal failure in this patient included hypovolemia, hypotension, and acute pancreatitis.


Cytokines, platelet activating factor and eosinophils in asthma. Understanding of the pathogenesis of asthma has increased considerably during the past few years. These advances were possible through scientific progress in three areas which contribute to this complex and multifaceted disease: (a) the much clearer understanding of eosinophil function; (b) the defining of lipid mediators in tissue inflammation and bronchial obstruction; and (c) the growing knowledge about the biological action of a new class of protein hormones, collectively called cytokines. In line with this, evidence has accumulated of how these components may interact with each other in providing the basis of inflammatory processes in asthma. Hence it seems appropriate to review the potential implications of this new information for the pathogenesis and therapy of this disease.


Radiological evidence of progression of bronchiectasis. Radiological evidence of progression of bronchiectasis was sought in a group of 84 consecutive adult patients admitted to a tertiary unit with a particular interest in the disease. Methodical comparison for each patient of the earliest and most recent chest X-rays (n = 84), bronchograms (n = 1) and thoracic computed tomography (CT) scans (n = 32) was performed. Fifteen patients (18%) were considered to show radiological evidence of progression of bronchiectasis, 14 on chest X-ray, two of whom also showed progression on CT scans, and in one patient on bronchography alone. The likelihood of finding evidence of radiological progression increased the longer the interval between examinations. Serial radiology allows identification of patients with progression of disease and indicates the need to review management strategy.


A high incidence of asthma and respiratory symptoms in 4-11 year old children. A study assessing the prevalence of respiratory symptoms in two primary schools in Birmingham, U.K. was performed. A questionnaire was delivered to pupils in both schools after which three open days were conducted in one of the schools, where probable asthmatics were identified and referred to their General Practitioner, Chest Clinic or a school asthma clinic. In this school 49% of responders and 52.9% in the control school were symptomless on questionnaire: 31% and 20.8%, respectively, had probable asthma, falling to 20% and 15.5% if a positive response to the question on recent recurrent wheeze was disregarded as indicating asthma. Using the total population as a denominator, the overall asthma prevalence was 20% which is significantly higher compared to previous English rates. Forty-two were seen at the Chest Clinic, 14 being followed for more than two visits. None were on regular anti-asthma treatment initially; 12/14 were taking prophylactic treatment on follow-up. In the two schools, 10.0% and 14.2% of responders were 'chesty' with 'colds' having no other typical asthmatic symptoms: these children should be studied further. This high incidence of respiratory symptoms in primary school children could represent a national trend or just a local increase.


Nebulized fenoterol causes greater cardiovascular and hypokalaemic effects than equivalent bronchodilator doses of salbutamol in asthmatics. The pulmonary and extrapulmonary effects of two doses of nebulized fenoterol (5 mg) salbutamol (5 mg) and ipratropium bromide (0.5 mg) at 60 min intervals were compared in nine patients with asthma in a double-blind, randomized study. Measurements of heart rate, blood pressure, electromechanical systole (QS2I), QTc interval, FEV1 and plasma potassium were made at baseline and at 15, 30 and 60 min after each nebulization. Both beta-agonists caused significantly greater inotropic (QS2I), chronotropic (HR), electrocardiographic (QTc) and hypokalaemic effects than ipratropium bromide (IB), with fenoterol being more potent than salbutamol. Fenoterol had no greater effect on FEV1 than salbutamol although both were superior to IB. Only the first four subjects had two doses as originally intended, because the second administration of fenoterol resulted in marked cardiovascular effects and hypokalaemia. The observed differences in extrapulmonary effects between fenoterol and salbutamol provide a plausible group of mechanisms which may explain the increased risk of death associated with fenoterol in severe asthmatics.


Chronic persistent cough: use of ipratropium bromide in undiagnosed cases following upper respiratory tract infection. The clinical effects of inhaled ipratropium bromide were studied in 14 non-smoking patients with persistent post-viral infective cough employing a controlled double-blind, cross-over trial. Patients were selected if they demonstrated no apparent underlying cause for their persistent cough after appropriate radiological and respiratory function tests including methacholine reactivity and bronchoscopic examination. Inhaled ipratropium bromide (320 micrograms day-1) produced significantly less day and night time cough (P < 0.05) with overall clinical improvement in 12 cases, five of whom had total resolution of their cough. We conclude that ipratropium bromide is an effective treatment in non-smoking adults with protracted cough following clinical upper respiratory tract infection.


Pulmonary tuberculoma and indications for surgery: radiographic and clinicopathological analysis. Tuberculomas of the lung are one of the more common lesions presenting a solitary pulmonary nodule, roentgenographically. We treated 36 patients with such nodules and describe here the radiologic-pathologic correlations and surgical treatment. In 21 patients, lung cancer was suspected preoperatively, based on radiographic findings of an ill-defined margin, pleural indentation and spicular radiation. Histologically, the tuberculous granuloma proliferated in the alveolar septa of the surrounding normal lung, often seen as a spicular radiation resembling lung cancer. In eight patients, tuberculoma was suspected because of radiographic findings of calcification and satellite nodules, and anti-tuberculous chemotherapy was prescribed for a few months. As this treatment was ineffective, surgical resection had to be done. Postoperative complications were nil and all of these patients are doing well at the time of preparation of this report. Anti-tuberculous chemotherapy was prescribed for 28 of 36 patients, postoperatively. We believe that surgical intervention is required for selected patients.


Pleural effusions: is thoracoscopy a reliable investigation? A retrospective review. In this paper, we consider the results of thoracoscopy in a busy thoracic unit where the referring physicians place their greatest emphasis upon simple standard investigation of pleural disease. Between 1985 and 1989 620 patients with a pleural effusion of unknown aetiology were referred to our thoracic medical unit. Initial investigations included aspiration of pleural fluid for cytology and culture, and blind pleural biopsy for histological examination. Recourse to thoracoscopy was only taken in the absence of a diagnosis or non-resolution of the patients symptoms and signs. Of these 620 patients only 48 (8%) remained without a diagnosis and were referred for thoracoscopy. Histological assessment of biopsies obtained at thoracoscopy revealed malignancy in 24 patients (50%) and benign conditions in 16 patients (33%). In eight patients (17%) no conclusive diagnosis was established; in this latter group, six patients continued with their symptoms and further invasive investigations revealed malignancy. In this setting where thoracoscopy was used as a last resort, the sensitivity for thoracoscopy was 83% and the specificity was 100% with a predictive value of a negative result being 25%. In conclusion, from our experience, the majority of pleural disease may be diagnosed using simple techniques but thoracoscopy can be very helpful in the more complex cases. Moreover, inconclusive histology following thoracoscopy is an indication for further investigation if the condition does not improve.


[Coastal urban malaria in Cotonou (Republic of Benin). Entomological study]. A one year entomological was carried out the survey in the coastal town of Cotonou to study the urban transmission of malaria. Three representative areas of Cotonou were chosen. The method adopted concerned night catches on human bait and dissection of A. gambiae s.l. The density of the vector (A. gambiae s.l.) fluctuated with the level of urbanization of the areas the annual aggressivity rates reached 1179 in the town center, 3666 in an the outskirts and 3363 in intermediate areas. Mean sporozoitic index was 1.7% and corresponding annual inoculation rate 46. In center of the town, transmission is seasonal and short, with a very high level. Sporozoitic index is 12% and the corresponding inoculation rate is 1.02. Outside of the town, transmission is seasonal and long: lasting 8 months. Transmission is imperceptible in the middle of the dry season and at the beginning of the long rain season. Malaria transmission into urban areas is generally low but in coastal like zones Cotonou, the intensity may be higher.


[Morbidity and severity of malaria attacks in carriers of sickle-cell trait]. The authors made a survey in a permanent high Plasmodium falciparum transmission area to compare frequency and severity of malaria attacks in children belonging to different haemoglobin types before 15 years; 291 young out-patients of the local infirmary and 467 outpatients of the hospital were examined. Diagnosis of malaria was inferred from clinical and parasitological criteria and subsequent evolution of the disease. Pathogenic threshold of parasitaemia was similar in all haemoglobin type groups of children and was about 3,000 parasite-infected red cells per mm3. Malaria was diagnosed more often among HbAA patients, than among other patients. Mortality rate in AA haemoglobin children was higher than 3% whereas in sickle cell trait carriers no death could be certainly attributed to malaria. The S gene rate was significantly weaker (p < 0.05) in subjects attacked by Malaria (5%) than in all other groups of children. In the endemic malaria areas the susceptibility of S gene carriers appears to be lesser than in AA haemoglobin children and could explain the paradoxically lower rate of mortality in this group.


[Plasmodium falciparum or P. malariae parasitemia in carriers of sickle cell trait in various Benin biotypes]. The prevalence of malaria and the frequency of gene S were surveyed in two different regions of Benin, savana and coastal lacustrine regions. In both regions, prevalence of malaria was not significantly different between Hb AA people and Hb AS people. Gene S prevalence was not modified by age, excepted for Hb SS which was not found in people upper than 25 years. In holoendemic area, i.e. lacustrine region, means of P. falciparum parasitaemia were significantly lower in Hb AS children than in Hb AA children. Sickle cell trait did not reduce the prevalence of malaria but seemed to decrease the level of parasitaemia.


[Urbanization and nutritional transition in sub-saharan Africa: exemplified by Congo and Senegal]. To evaluate the impact of urban life-style on nutritional status, body mass index (BMI) of mothers and indices of malnutrition of preschool children were calculated in four representative surveys in two rural areas and two main cities of Western and Central Africa. Mean BMIs were similar in both urban settings and were significantly higher than those of rural mothers. Distributions shifted significantly towards values over 25 kgs/m2 in towns, although, values lower than 18.5 were still present. Therefore chronic energy deficiency, largely prevalent in many rural areas of Africa, remains important in cities, where obesity also appears to have become a public health concern. Wasting was rare in young urban children, but the prevalence of stunting, although lower, indicates the persistence of nutritional deficiencies. This situation of nutritional transition generates a double burden to already limited health finances and requires an appropriate educational policy.


[Anemia at delivery in Lome (Togo): prevalence, risk factors and consequences in newborn infants]. A prevalence study was carried out on 125 mothers and their newborns in Lome (Togo): at delivery 48% of the mothers and 30% of the newborns were anaemic according to WHO criteria. Iron deficiency was the major determinant of anaemia in the mothers, as three out of four showed at least one biochemical indicator of iron deficiency. Folate deficiency was detected in 68% of the mothers but did not influence their haematological parameters. Severe iron deficiency in the mothers (serum iron < 7 mumol/l) was associated with a decrease in serum iron in the newborns, thus demonstrating an impaired iron transfer to the fetus. Folate supplementation of the mothers during pregnancy improved their newborn's folate status. A systematic ferro-folic supplementation is needed during pregnancy and would be beneficial to both mothers and newborns. Supplements could be given to women at prenatal care clinics. Attendance in these centers by 98% of pregnant women in Lomé allows us to anticipate a good coverage for such an intervention.


[Man-water contacts and urinary schistosomiasis in a Mauritanian village]. For the period September to December 1985, 1226 water contacts were recorded during 8 days of direct observation. Various activities were analysed in order to determine their responsibility in transmission. An index of exposure, allowing for duration of contact, body surface exposed and infectiousness of the water was calculated for each contact. Domestic contacts, primarily female, represented 62% of the observations but only 15% of total exposure. Conversely, contacts for recreational purposes mainly involved young boys and accounted for 14% of the observations and 70% of total exposure. Between 6 and 20 years of age the mean index of exposure by contact was higher in males than in females. Changing water contact behavior seems to be an unrealistic means of preventing transmission in the community studied. The most appropriate strategy of control would appear to be selective treatment of heavily infected individuals.


[Polyneuropathies associated with lymphoplasmacytic dyscrasias: the clinical and instrumental characteristics of a 38-patient case load]. A varying percentage from 40 to 60% of patients having lymphoplasmacytic dyscrasias with a monoclonal component shows a clinical or subclinical polyneuropathy. From a different viewpoint, a monoclonal gammopathy has been detected in 10% of patients effected by chronic idiopathic polyneuropathy. A case study of 38 patients with lymphoplasmacytic dyscrasia subjected to clinical, immunohematological and electrophysiological examination revealed a high prevalence of polyneuropathy (84%), mainly axonal (72%) and often subclinical. The neuropathy was evenly distributed between patients having malignant and benign lymphoplasmacytic dyscrasias. No statistically significant correlation was found between the presence of neuropathy and the main clinical and immunohematological data. This supports the concept that the pathogenesis of polyneuropathy associated with lymphoplasmacytic dyscrasias may be multifactorial. Nor can it be ruled out that the paraprotein may in fact be secondary to the polyneuropathy or sometimes a simple coincidence.


[The soluble IL-2 receptor in malignant hemopathies]. The increase in the serum levels of the IL-2 receptors is due to its release both in vivo and in vitro from activated cells or neoplastic cells expressing it constitutively. The diagnostic, prognostic and physiopathologic significance of the sIL-2R was investigated by testing the serum of 271 haemopathic patients in various stages of the disease. In HCL the elevated sIL-2R level has a diagnostic value. In HD the sIL-2R level appears to be directly correlated with the extent of the disease and is equally important in the follow up of patients with HCL, NHL, HD, AL and MDS, where the serum level of the soluble receptor is usually associated with the biological and clinical activity of the disease. Unlike other B lymphoproliferations, patients with Multiple Myeloma on average show only slightly elevated levels of soluble receptor with no significant differences related to the stage or evolution. As for the chronic myeloproliferative disorders, we found only slightly elevated values in ET and PV, with frankly pathological values in CML during a blastic crisis or in the accelerated phase and in MFI during the clinically active phase of the disease.


[Ebstein's tricuspid anomaly and Down's syndrome. A clinical case report]. The authors describe a case of a 55 years old white man with Down's syndrome in whom an Ebstein's anomaly of tricuspid valve was casually diagnosed. The peculiarity of this association was searched in the literature, trying to explain, at the same time, the poor symptomatology. The role of echocardiography in the diagnosis of Ebstein's anomaly is emphasized.


[Horton's bitemporal arteritis. A case report]. The authors report a case of Horton's arteritis in a woman (aged 76) presented some peculiarities. The arteritis simultaneously interested both superficial temporal arteries; in a second time caused a bilateral "claudicatio masticationis"; induced a worsening of extrapyramidal symptoms from which the patient had been suffering for many years, due to the possible involvement also of the endocranial arteries. The histologic picture on a biopsy of the left temporal artery was typical of the Horton's disease, but without giant cells; the authors suppose that this is due to a therapy with FANS administered before carrying out the biopsy. The patient was treated with cortisone per os at a low dosage, causing a quick regression of the symptoms and a normalization of the laboratory tests. Also this result differs from what most authors have observed: they maintain that Horton's arteritis has become rather resistant to the cortisone therapy and required high dosage for a very long time. The authors maintain that precocity of diagnosis and an immediate treatment can influence in a significant way the course of the disease.


[Totally implantable systems for long-term chemotherapeutic treatment]. The authors report their own experience with completely implantable systems for the long-term chemotherapeutic treatment, compared with a review of modern literature. The experience even if low demonstrates a good methodology in reducing the number of complications associated with the use of percutaneous catheters.


[Recent findings on the pathogenesis and therapy of anemia in chronic kidney failure]. Recent studies showed that the blood BFU-E, when subtracted from the uremic milieu, normally responds to the stimulating factor produced by T lymphocytes. The serum of uremic patients inhibits the in vitro growth of normal BFU-E, however, the inhibition is almost completely reversed by hemodialysis. These data allow to understand why the therapy with erythropoietin relieves the anemia of CRF. Uremic T lymphocytes fail to stimulate the BFU-E growth. Normal T lymphocytes are inhibited by uremic serum and the hemodialysis does not correct the defect. Lymphopenia, decreased number of both T4 and T8 lymphocytes and low T4/T8 ratio were found in 50% of patients. Cimetidine was still able to increase the burst-stimulating activity of uremic T lymphocytes through inhibition of the suppressor T subset. In conclusion, one can say that in CRF T8 lymphocytes are normal and that uremic toxins decrease both number and function of T4 lymphocytes. The deficiency of BPA appears to significantly contribute to the pathogenesis of the anemia of CRF. The experience from our and other Institutions shows the effectiveness of the recombinant human erythropoietin in relieving the anemia of CRF, notwithstanding the hematological milieu is highly modified by uremia.


[Glanzmann's thrombasthenia: a rare example of an integrin deficit]. Glanzmann's thrombasthenia is an autosomal recessive life-long bleeding disorder, originated from a quantitative or qualitative defect of the major platelet membrane receptor: the GPIIb/IIIa complex. The GPIIb/IIIa complex is a calcium-dependent heterodimer, belonging to the integrin superfamily. The complex of activated platelets can bind fibrinogen, von Willebrand factor, fibronectin and vitronectin, which are proteins playing an important role in platelet adhesion and aggregation. Thrombasthenic platelets are deficient in GPIIb, GPIIIa and GPIIb/IIIa complex; however, platelets from few thrombasthenic patients content near-normal amounts of functionally abnormal complex. The GPIIb/IIIa quantitative or functional defect leads to defective platelet hemostatic plug formation. Hemorrhagic symptoms consist of purpura, gingival hemorrhage, menorrhagia and epistaxis. Some cases of Glanzmann's thrombasthenia have been characterized at the molecular genetic level. Molecular abnormalities include: GPIIb or GPIIIa partial gene deletion, GPIIIa gene insertion, a point mutation resulting in an amino acid substitution within the GPIIIa molecule. In spite of the contemporary reduction of both GPIIb and GPIIIa in most cases of Glanzmann's thrombasthenia, it appears that a molecular abnormality affecting only one of the glycoprotein genes may result in a thrombasthenic phenotype.


[The endothelial cell: a cell structurally simple but functionally very complex]. Vascular endothelium is morphologically similar to a simple squamous epithelium that lines the internal surface of vessels. These cells, even if structurally simple, are functionally very complex. They regulate vascular tone by producing vasodilators such as prostacyclin and nitric oxide and vasoconstrictors such as endothelin. In addition, endothelial cells express activators and inhibitors of coagulation, fibrinolysis and platelet aggregation. Alterations of the functional equilibrium of endothelial cells might be related to the development of many cardiovascular diseases.


Exposure to ozone and erythrocyte osmotic resistance in the rat. In order to learn the biological effect of photochemical oxidants on living bodies, we exposed newborn and adult rats, of both sexes, to ozone at a concentration of 0.25 ppm, which can be encountered in an urban environment, and then measured the osmotic resistance of their erythrocytes. The results of experiments using newborn rats indicated a positive increase in the osmotic resistance of erythrocytes in whole blood following ozone exposure for 4 weeks. An increase in the osmotic resistance of erythrocytes in the top part obtained by centrifugation was observed following ozone exposure for 12 weeks. This tendency was especially evident among male rats. On the other hand, no increase in the osmotic resistance of erythrocytes was recognized in the adult animals which had been exposed to the same concentration of ozone for 18 months.


In vivo glycogen and lipid synthesis by various tissues from normal and metformin-treated KK mice. The incorporation of [14C] U-glucose into glycogen by liver, kidney and skeletal muscle and that into total lipid by adipose tissue was studied in noninsulin-dependent genetically diabetic KK mice treated either with metformin (50 mg/kg) or water for 10 weeks. Results show that glycogen synthesis was increased in the skeletal muscle from metformin-treated mice. Liver and kidney glycogen synthesis was not affected by metformin. Glucose incorporation was significantly increased into lipid in metformin-treated mice. Insulin administration stimulated lipid synthesis in adipose tissue, but had no effect on muscle glycogen synthesis in metformin-treated mice. These data suggest that chronic metformin treatment potentiates the endogenous insulin action on glucose in skeletal muscle and adipose tissue, and thus confirm our previous results of increased glycogen synthesis by skeletal muscle in metformin-treated KK mice.


Pseudocholinesterase in alloxan-diabetic rats. Pseudocholinesterase (PChE) activity in plasma, liver, pancreas and adipose tissue of alloxan-diabetic rats has been determined. 48 hours after the induction of diabetes an increase of PChE activity in plasma and a decrease in liver and adipose tissue was observed. After that, the activity in plasma remained at the same level, in the liver it slowly rose to the control values and in adipose tissue significantly increased beyond the control values. Significant changes of PChE activity in the pancreas were not noticed. Female rats had higher PChE activity in plasma and liver than the male, but in the pancreas and adipose tissue there were no sex differences. On the basis of our results it seems possible to propose that PChE is synthesized also in adipose tissue, but the synthesis is regulated by different mechanisms than those in the liver.


Synergism of cyclosporin A and phospholipase inhibitors in protection against lethal injury to rat hepatocytes from oxidant chemicals. Cyclosporin A and phospholipase inhibitors block the oxidant-induced mitochondrial permeability transition. Here, protection by these agents against cytotoxicity of oxidant chemicals was evaluated in rat hepatocytes. The combination of cyclosporin A and a phospholipase inhibitor (trifluoperazine, mepacrine, or dibucaine), but neither alone, substantially protected against lethal injury from 0.5 mM iodoacetate but had little effect against iodoacetate plus 2.5 mM KCN. Against 100 microM t-butylhydroperoxide, cyclosporin A and trifluoperazine protected only if fructose was present. Cyclosporin A plus phospholipase inhibition protected slightly against 2.5 mM cystamine, but actually increased lethal injury after 100 microM menadione. These effects are consistent with an oxidant-induced mitochondrial permeability transition that accelerates cell killing after iodoacetate and t-butylhydroperoxide.


Membrane-associated phospholipase A2 stimulates DNA synthesis in two murine fibroblasts. The effect of human membrane-associated phospholipase A2 (M-PLA2) on DNA synthesis in fibroblasts was examined in order to clarify its biological function. Human M-PLA2 added exogenously to the fibroblast cell lines Swiss 3T3 and BALB/3T3 was found to augment their DNA synthesis. This stimulation of DNA synthesis was not affected by treatment with indomethacin. Our results suggest that human M-PLA2 itself has a mitogenic effect on fibroblasts independent of arachidonate products such as prostaglandins.


Cardiovascular actions of NG-methyl-L-arginine are abolished in a canine shock model using high-dose endotoxin. This study was designed to test the hypothesis that endotoxin-induced hypotension is caused, in part, by increased endothelial synthesis of nitric oxide and that inhibition of nitric oxide synthesis with NG-monomethyl-L-arginine (NMA) would reverse the cardiovascular actions of endotoxin. A high dose of endotoxin (1.5 mg/kg, i.v.) was administered by rapid intravenous infusion to pentobarbital-anesthetized dogs. Endotoxin caused rapid and profound reductions in cardiac output and mean arterial pressure; systemic vascular resistance, however, was unaltered except for a transient increase. Coronary and mesenteric blood flows were reduced. NMA (30 mg/kg, i.v.) given 60 min after endotoxin administration, had no significant effect on cardiac function or systemic vascular function except for a transient increase in cardiac output and decrease in systemic vascular resistance. This same dose of NMA given to dogs not receiving endotoxin caused systemic vasoconstriction, increased arterial pressure and decreased cardiac output. These results suggest that increased nitric oxide production is not a primary factor causing endotoxin-induced hypotension and that nitric oxide does not modulate vascular tone following administration of high doses of endotoxin.


The "anesthetic" effect of alcohols and alkanes in Caenorhabditis elegans (C.e.). In order to help validate the free-living roundworm, C.e., as a simple model to study the mechanism of general anesthesia, we demonstrated that homologous series of alcohols and alkanes produced a reversible "anesthetic" effect in these worms as in other animals. Also, as in other animals, the potency of these compounds was directly related to their respective lipid solubilities within each series. The alcohols were much more potent than the alkanes even though the latter were much more lipid soluble than the alcohols. The cut-off point (a decrease followed by a loss of activity) for these compounds occurred at about C9. In mice, the alcohols had an effect analogous to that seen in C.e. but the alkanes were inactive after intraperitoneal injection.


Oxidative metabolism of flunarizine in rat liver microsomes. The oxidative metabolism of flunarizine [1-[bis(4-fluorophenyl)-methyl]-4-(3-phenyl-2-propenyl)piperazine, FZ] to 1-[bis-(4-fluorophenyl)methyl]piperazine (M-1), 1-[bis(4-fluorophenyl)methyl]-4-[3-(4'-hydroxyphenyl)-2- propenyl]piperazine (M-2) and 4,4'-difluorobenzophenone (M-3) has been studied in liver microsomes of Wistar and Dark Agouti (DA) rats. Kinetic analysis demonstrated a sex difference (male > female) in the formation of M-1 and M-3, but not in that of M-2 in Wistar rats. Comparison of the kinetic data of FZ with those of cinnarizine [1-(diphenylmethyl)-4-(3-phenyl-2-propenyl)piperazine, CZ], a prototypic and unfluorinated drug (Kariya et al., Biochem. Pharmacol., in press) revealed that the formation clearances (Clfs) estimated by Vmax/km for the ring hydroxylated metabolites of FZ and CZ are higher than those for the N-dealkylated metabolites of these drugs in female rats. Furthermore, the introduction of two fluorine atoms to CZ (forming FZ) decreased the Clfs for most of metabolites, especially for the N-dealkylated product, M-3. The formation of the metabolites from FZ was suppressed by carbon monoxide and SKF 525-A, and only the ring hydroxylation forming M-2 was significantly lower in female DA than in female Wistar rats. These results suggest that the microsomal oxidation of FZ is mediated by cytochrome P450, and that a cytochrome P450 isozyme(s) belonging to the CYP2D subfamily is involved in the ring hydroxylation of FZ forming M-2.


A simplified gelatin tolerance test to evaluate gastric and pancreatic proteolytic activities. To evaluate gastric, pancreatic and intestinal proteolytic activities, a gelatin tolerance test was standardized, and its clinical significance was evaluated in 16 normal subjects and 25 patients who had undergone gastrectomy or had a pancreatic disease. A 15g gelatin dose given to normal subjects resulted in an increase of serum-free and peptide-bound hydroxyproline (HYP), with the peak values being observed at 1 hr and 2 hr, respectively. In patients who had malabsorption, both peak levels of serum-free and peptide-bound HYP were decreased, and the peak value of free HYP was more frequently lower than that of peptide-bound HYP. The determination of serum free HYP after oral administration og gelatin was significantly correlated with BT-PABA test. These results indicate that usefulness of determining serum-free HYP for the gelatin tolerance test as a simple reliable test for digestion and absorption of proteins.


Reimbursement of biotherapy: present status, future directions--perspectives of the third-party payer. The mission of insurers is to provide defined financial support for health care therapies deemed appropriate for use in specific clinical situations. In the current health care financial crisis, insurers are faced with keeping costs to a minimum and premiums reasonable. While insurers wish to finance the best available treatment, it is not always fiscally responsible or realistic to fund care provided in investigational therapies. The Blue Cross and Blue Shield Association (BCBSA), as the national coordinating body for local Blue Cross and Blue Shield Plans, assesses the status of new technologies, such as the biotherapy of cancer, through its Technology Evaluation and Coverage (TEC) Program and its Medical Necessity Program. Fundamental to both programs is whether a technology is effective: Does it improve health outcomes? And, if it does, what are its appropriate conditions of use? New technologies demonstrated by clinical research to improve health outcomes and found consistent with other related criteria are considered eligible for coverage by the TEC Program. New technologies not yet established as effective by clinical research and approved through the TEC Program are considered investigational. Most Blue Cross and Blue Shield Plan contracts exclude coverage for such investigational technologies. Most plan contracts also have medical necessity clauses, whereby only medically necessary technology uses are covered. Central to this is clinical research on appropriate medical conditions of use--the focus of the Medical Necessity Program. Valid conclusions on whether a technology works and where and when it works the best presuppose well-designed scientific studies.(ABSTRACT TRUNCATED AT 250 WORDS)


Patterns of self-care needs and interventions related to biologic response modifier therapy: fatigue as a model. Many patients receiving biologic response modifier (BRM) therapy experience fatigue as a significant and, at times, dose-limiting side effect. For this reason, a multiinstitutional pilot study was designed to collect data about the needs and self-care interventions of patients who had undergone at least one prior treatment with a BRM and had experienced fatigue as a symptom. Information was also obtained on the extent to which the needs and self-care interventions identified by patients compared with those perceived by their family members and nurses. Of the 16 patients who participated in the study, seven were being treated with interleukin-2 (IL-2), eight with interferon alfa (IFN-alpha), and one with tumor necrosis factor (TNF). The study found no significant correlation between the degree or duration of fatigue and the BRM or dosage administered. Not surprisingly, patient and family member responses correlated fairly well. However, in several parameters, including the degree and duration of fatigue, nurses' perceptions did not correlate at all with those of the patient. While 46% of nurse responses matched those of the patient as to useful self-care interventions, only 17% of nurses accurately identified factors or events that patients perceived as worsening fatigue. Further, there was no correlation among patient, family member, and nurse responses on interventions that could be used by others to help the patient cope with fatigue. The overall results of this pilot study indicate that nurses need to be more attuned to assessing fatigue as a side effect of BRM therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


Symptom assessment and management of outpatients receiving biotherapy: the application of a symptom report form. Patients treated with high doses of biologic response modifiers (BRMs) often require hospitalization for monitoring and management of toxicities. However, with careful symptom assessment and management, patients can be treated safely with low-dose, long-term BRM regimens in an outpatient setting. Quantifiable, consistent, and accurate symptom assessment sets the groundwork for regular, effective symptom management. A symptom report form or nursing care flow sheet is a helpful daily assessment tool for recording and quantifying the most common side effects experienced by patients receiving BRMs. Parameters can be established and converted to a numerical code to provide a consistent guide for evaluating a patient's toxicities during treatment. Additional space on the report form allows for narrative comments about symptoms or concerns not addressed elsewhere. To be safe and effective, outpatient therapy of this kind requires excellent patient education and compliance, with easy access to a health care professional when necessary.


Reimbursement of biotherapy: present status, future directions--perspectives of the hospital-based oncology nurse. Cancer is a set of diseases that affects one in four Americans and which is too often fatal. For many types of cancer, there is no fully satisfactory treatment. Therefore, investigational therapy provided in a formal, peer-reviewed clinical trial can present the best available treatment for patients with life-threatening malignancies. At a time when clinical cancer research is at a particularly promising point and multiple new biologics are on the horizon, the traditional partnership of the federal government, the pharmaceutical industry, private institutions, and insurers is threatened. The issue of increasing concern is the impact of changes in third-party reimbursement policy on cancer clinical investigation. Although its incidence, magnitude, and geographic distribution have not been specifically defined, this issue is generating serious concern and intense national debate. The growing denials of patient-care costs related to clinical trials serve to threaten patient access to potentially efficacious therapy. The implications of this reimbursement issue are tremendous for both clinical investigation and patient care. The refusal to pay for patient-care costs related to investigational therapy may adversely influence clinical trial design and will certainly affect the speed with which new therapies are developed. When insurers refuse to cover these clinical costs, only affluent individuals have access to the most promising treatments. Nursing actions can positively affect the reimbursement debate. Nurses must develop cost-efficient plans of care and the means to evaluate their effectiveness. Educating patients about and assisting them with reimbursement issues maximizes their prospects for receiving appropriate investigational therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


Reimbursement of biotherapy: present status, future directions--perspectives of the office-based oncology nurse. Economic forces are stimulating cost sensitivity and the need for clinical efficiency in medicine. The federal government has led the way with Medicare reform, and cost-containment efforts are evident in all health care payer programs. More and more, the office-based oncology nurse is involved in reimbursement issues as both a colleague and a patient advocate. Creative solutions to the challenges of reimbursement require knowledge of the issues, familiarity with patient-specific therapies, and recognition of unique cost and billing issues. Biologic agents frequently used in office-based oncology practice are easy targets for reimbursement denials because of regulations against their investigational status, "off-label" use, method of administration, and relatively high cost. Such agents include interferon, erythropoietin (EPO), granulocyte and granulocyte-macrophage colony-stimulating factors (G-CSF and GM-CSF, respectively), and interleukin-2 (IL-2). Reimbursement decisions are often characterized by inconsistency and uncertainty, and rulings are based not only on law, but also on interpretation. The need for clarification often opens a window for negotiation for the complex reimbursement issues associated with biotherapy. In addition to thoroughly determining cost and accurately assigning appropriate Current Procedural Terminology (CPT) codes, office-based oncology nurses can pursue various strategies to help their patients and practices obtain reimbursement of biotherapy. Chief among these is educating third-party payers on the appropriateness and necessity of newer treatment modalities. In individual cases, documentation of the scientific data, clinical outcomes, and cost benefits supporting a treatment decision almost always gains reimbursement.(ABSTRACT TRUNCATED AT 250 WORDS)


[Pulmonary embolism: which diagnostic approach?]. The recent literature dealing with the diagnosis of pulmonary embolism is reviewed. Clinical signs, electrocardiogram and arterial blood gases analysis are not very helpful whereas a normal level of blood D-dimers makes the diagnosis of pulmonary embolism very unlikely. Lung scanning must be interpreted carefully in parallel with chest radiography. It is most useful if the pattern is either normal or of high probability. All intermediate scans are inconclusive and need a pulmonary arteriogram. Another option in patients with good cardiorespiratory reserve is the use of repeated non-invasive investigations of the lower limbs.


[Creation of a planned autologous transfusion unit in a hospital distinct from a transfusion center]. Autologous blood predonation now widely used for programmed surgery has been recently regulated by a law from january 9 1992. It establishes that the procedure must be performed under the responsibility of an accredited blood transfusion center. This work discloses that the establishment of an autotransfusion unit in a hospital endowed with an active surgical program is feasible, the major objective being improvement of the quality of life of elderly patients with invalidating diseases. We report the one year experience of a collaboration between the Braine-l'Alleud-Waterloo hospital and the Red Cross Erasme-Brabant Wallon blood transfusion center. Eighty patients enrolled in programmed surgery consisting mostly in orthopedic surgery have given 187 units of packed red blood cells and fresh frozen plasma. Sixty two patients (83%) were transfused solely with autologous blood. Tolerance was excellent as only 4 slight malaise episodes were observed. Our study also indicates that programmed predonation should be proposed mainly to patients for whom the prescribed blood is effectively transfused. Creation of such autotransfusion units in hospitals distinct from a blood transfusion center is feasible with the deep collaboration between physicians in charge of the patients, the hospital laboratory and the blood transfusion center.


[Persistent and recurrent hyperparathyroidism following parathyroidectomy]. In a series of 416 parathyroidectomies for primary or secondary hyperparathyroidism, 19 were reoperations for persistence (17 cases) on recurrence (2 cases) of the disease. (1) Preoperative localisation studies were useless in half of the cases. (2) In re-explorations, 72% only of parathyroid glands were discovered, 46% of them in ectopic locations. In reoperations for primary hyperparathyroidism, 56% of cases had more than one pathological gland. (3) Long term results have been less satisfactory after re-explorations than after the first operations.


[Current aspects of neurosyphilis: therapy-resistant cases with high-dosage penicillin?]. The occurrence of atypical and abortive cases of neurosyphilis in the last two decades has tended to make this rather infrequent, treatable condition difficult to diagnose; therefore it is advisable to perform large-scale serologic tests among neurologic and psychiatric patients. The diagnosis is confirmed by the detection of treponema antibodies in the central nervous system (TPHA index, ITpA index). When serologic test results are positive, secondary diagnostic criteria are provided by pathologic CSF findings (elevated cell count, humoral immune reaction). The course of the disease and the CSF parameters are favourably influenced by appropriate therapy. Over the last two decades, there has been an increase in the number of reports of cases refractory to the low-dose penicillin treatment recommended by the WHO and the CDC until two years ago. There have also been reports of treponema resistance in association with this therapeutic strategy and furthermore of failure to detect CSF concentrations of penicillin capable of eradicating the spirochetes. Counter measures have been taken worldwide in the last few years in the form of high-dose penicillin therapy (approx. 20 million IU/day for 2 to 3 weeks), in the hope that this would eliminate the resistance. We observed two patients treated with 18 and 24 million IU penicillin for 10 and 14 days respectively. The pathogens remained resistant, and the disease continued to progress. In one of these cases, immunologic processes might have been predominantly responsible for the resistance to the treatment, while in the other resistant pathogens can be assumed to be responsible. Repeated courses of treatment with higher doses of penicillin and/or with ceftriaxone finally succeeded.(ABSTRACT TRUNCATED AT 250 WORDS)


[Localization of level of lesions in internuclear ophthalmoplegia through assessment of masseter and blink reflex]. The masseter and blink reflexes were investigated in 100 patients with internuclear ophthalmoplegia due to multiple sclerosis (58 patients) or lacunar brainstem infarction (42 patients). In unilateral internuclear ophthalmoplegia, 38 of 60 patients (63.3%) had masseter reflex abnormalities, two patients (3.3%) showed changes of the blink reflex R1 component, and 13 patients (21.7%) combined alterations of the masseter reflex and the blink reflex R1 component. 46 (86.8%) of these 53 patients with electrophysiological abnormalities had unilateral changes, which were ipsilateral to the medial longitudinal fasciculus lesion in 42 patients (91.3%). In bilateral internuclear ophthalmoplegia, 24 of 40 patients (60.0%) had abnormalities of the masseter reflex, two (5.0%) showed changes of the blink reflex R1, and nine (22.5%) combined alterations of the masseter reflex and the blink reflex R1 component. 20 (57.1%) of these 35 patients with electrophysiological abnormalities had bilateral changes. Thus, masseter reflex abnormalities indicating midbrain lesions were seen in 63.3% and 60.0%, respectively, of unilateral and bilateral internuclear ophthalmoplegia. Blink reflex R1 component changes with or without impairment of the masseter reflex indicating rostral pontine to midpontine lesions occurred in 25.0% and 27.5%, respectively. These figures correspond to the results of postmortem examinations and to theoretical considerations based on the length of the medial longitudinal fasciculus.


[Romantic origins of electrophysiology]. Research on static electricity and its effects on the human body date back to the invention of the electrizing or Wimshurst machine and the Leyden jar of 1743 and 1746. Such experiments often served as social pastimes, but they yielded many publications on medical aspects of static electricity. Attempts to explain the 'life force' of the vitalists and the old concept of the active principle of the nervous system, the 'spiritus animales', as electrical phenomena were unsuccessful because of the skeptic comments of leading experimental scientists such as Albrecht von Haller. When Mesmer reinvented 'animal magnetism' in 1776 as a fashionable term for treatment by suggestion, he appropriated theoretical, technical and social methods from the established ways of the experiments on static electricity. Therefore, the scientific character of Luigi Galvani's investigations was already compromised by his term 'animal electricity' when he published his famous 'Commentarius' in 1781. Volta in Pavia turned against Galvani, proving that galvanic currents are produced by metals alone, and rejecting 'animal electricity'. Volta's doctrine prevailed over Galvani's school after Volta's breakthrough with his pile, or battery, until Galvani's ideas were rehabilitated by Nobili, who in 1828 measured the 'frog current' with his galvanometer. This led to a flurry of bizarre experiments on rows of half-dismembered animals and severed parts of human cadavers. Johannes Müller in Berlin, who, with his students, established new principles of biology and neurology, asked Du Bois-Reymond to study these experiments. Du Bois-Reymond found that measurements of muscle currents in intact animals were more useful, and he compared them with his own observations on electric fishes.(ABSTRACT TRUNCATED AT 250 WORDS)


[Motor evoked potentials. Physiology, indications, safety aspects]. The motor cortex and the proximal part of the peripheral nervous system can be stimulated by a rapidly changing magnetic field. The procedure is simple, painless and safe. It allows to quantify the function of the most rapidly conducting corticospinal fibers. It is possible to detect subclinical lesions. The use of this technique for the study of lesions of the proximal parts of the peripheral nervous system is as of yet largely experimental.


[Gait analysis--a prerequisite for general movement analysis]. Our system for analysis of gait is a combined visualisation of angular velocities in any joint with dynamic measurements of pressure at the sole while walking in a natural surrounding. Infrared reflectors are attached to chosen parts of the body. Their position in space is sampled over time and transferred to an easily manageable personal computer that visualizes motion in progression as angular velocities. Pressure at the sole is measured by pressure sensors. The magnitude of force is expressed as calibrated circular areas following the dynamic course of gait. The system is accurate and easily operated. The time necessary for measurements and evaluation of data as well as the costs render an application in daily clinical work acceptable.


[Epilepsy in middle and older age].

[Epilepsy in middle and older age]. Single fits and epileptic illness are more frequent in advanced age; their occurrence reaches the same frequency as in the neonatal period. Their origin and the possibilities of treatment are tightly connected to the process of ageing, a fact that requires special consideration. The main reasons for new epileptic attacks in the group of age 65 or more are structural changes, i.e. ischemic infarctions, tumors and atrophic involution. For certain patients, the use of antiepileptic drugs may be limited by unavoidable side effects. Complete cure of the attacks in advanced age is rare, but with a well selected treatment sufficient control is often possible.


Genetics of transitional cell carcinoma. Several models of genetic events which define the origin and progression of human tumors have been elucidated over the last several years. These models suggest that the study of tumors at the level of both the chromosome and the gene can be useful in elucidating molecular events in tumor progression and in determining the biologic behavior of individual tumors. The genetics of transitional cell carcinomas are reviewed with emphasis on potential mechanisms of tumorigenicity and the clinical utility of genetic markers.


Biology of metastasis: clinical implications. Bladder tumor has a spectrum of neoplastic activity. Some behave in a benign fashion, and others are highly aggressive and lead rapidly to metastatic disease and death. The processes of metastasis can be described as a sequence of interrelated steps. The processes involve 1) tumor cell adhesion to basement membranes, 2) the degradation of basement membranes, and 3) the migration of tumor cells through the destroyed stroma into blood and lymphatic vessels. Each of these processes involves the expression of molecular factors unique to tumor cells. With better understanding of the molecular basis of these factors, novel prognostic and potential therapeutic agents can be generated and applied to the clinical arena.


Critical review of the models to study the biologic progression of bladder cancer. For transitional cell carcinoma of the bladder, clinical data indicate that invasive, metastatic tumors can arise through at least two different progression pathways. The majority of invasive, metastatic bladder neoplasms clinically present de novo, i.e., the patients have no history of malignant bladder disease. This implies that the highly malignant tumor cells either arise de novo or have undergone a rapid progression. Alternatively, a considerable fraction of patients with superficial bladder cancer process to invasive disease after a history of relatively benign superficial TCC. The molecular and cell biological basis of tumor progression is only poorly understood. Clearly, a better understanding of this progress could have profound clinical implications, since patients with superficial TCC with a high risk for progression would have to be treated more aggressively. We discuss the problems that are associated with tumor biological studies on early steps in the progression of TCC, especially from a "model system point of view."


Experimental models of histogenesis and tumor cell heterogeneity in bladder cancer. The histogenetic relationships between the subtypes of bladder cancer are not known. Each common pattern (transitional cell carcinoma, adenocarcinoma, and squamous carcinoma) can exist independently, although they may coexist in primary or metastatic bladder cancers, and tumors that are predominantly composed of transitional cell carcinoma may have regions of squamous or glandular differentiation. Morphologically identical tumors exhibit marked variation in their natural history and response to treatment. To study these aspects of the biology of human bladder cancer, a series of cell lines have been established and characterized as xenografts and in tissue culture. These studies have shown a likely common origin for transitional cell carcinoma, adenocarcinoma, and squamous carcinoma of the bladder. Morphologically similar xenografts and cell lines in vitro have shown a broad range of functional heterogeneity, including ultrastructure, tumor marker production, ploidy, cell surface characteristics, and response to chemotherapy. These are useful models of heterogeneity of response to treatment with established and new cytotoxic agents.


Growth factors and bladder cancer: clinical implications of the interactions between growth factors and their urothelial receptors. Growth factors (GFs) are a class of proteins that bind to specific cell surface receptors (GF-Rs), inducing a variety of responses including mitosis, in susceptible target cells. Abnormal production, expression, and/or function of GFs or GF-Rs can result in unregulated growth, the hallmark of malignant transformation. This chapter reviews those GFs/GF-Rs that have been linked to human bladder cancer. It focuses particularly on one [epidermal growth factor (EGF), which is excreted in urine in high concentrations] and its possible role in the development and growth of urothelial malignancy. Potential clinical applications in diagnosis, staging, prevention, and treatment are discussed.


Cell surface differentiation antigens of normal urothelium and bladder tumors. Bladder cancer ranks as the third most common malignancy among men and tenth among women. Superficial transitional cell carcinomas (stage Ta, Tis, and T1) account for approximately 70-80% of these tumors, while the remaining 20-30% are invasive (T2, T3, and T4). Approximately 70% of superficial tumors will have one or more recurrences, with 25% of these expressing a higher histologic grade and 10-15% subsequently developing invasive and/or metastatic disease. The detection and prediction of tumor recurrence and/or tumor progression is crucially important if timely and appropriate therapy is to be instituted. Conventional histopathologic evaluation usually provides definitive diagnosis upon which therapeutic planning is based. However, at present there are no more reliable morphologic indicator to identify which individuals will have recurrent disease or who will progress to invasive and/or metastatic cancer. Recent advances in tumor biology have identified markers that are good candidates for clinical applications in early tumor detection, as well as for the stratification of patients with like-appearing morphological lesions with different biological and clinical behavior. The ultimate goal is to develop predictive assays that would segregate patients with high probability of failures versus patients who would be cured by localized modes of therapy.


Flow cytometry for detection and evaluation of urinary bladder carcinoma. Flow Cytometry (FCM) DNA assays of bladder irrigation specimens are now recognized as a clinically useful and reliable means of detecting and monitoring carcinoma of the bladder. This technique, which identifies carcinoma by the presence of an aneuploid population of cells, can be carried out on specimens obtained in an outpatient or hospital setting and is easily performed in any medium-sized laboratory. It is most sensitive to superficial and high grade tumors. Overall, nearly 80% of superficial carcinomas of bladder will have positive flow cytometry, comparing very favorably with conventional cytology. Until now, the widest clinical application of FCM has been in monitoring the conservative treatment of stage 0-1 flat and papillary carcinomas, but newly developed dual parameter measurements are capable of quantifying proliferative activity, oncogene expression, growth factor receptors, and other cellular features that may better characterize the biologic potential of these tumors and can be expected to aid in the selection and timing of treatment.


Expression of blood group antigens in bladder cancer: current concepts. Blood group antigens are a group of carbohydrate structures bound to membrane lipids or proteins of erythrocytes and certain epithelial tissues including urothelium. The Lewis antigens are structures that are genetically and biochemically related to the ABO blood group. The ABO and Lewis blood group systems are differentially expressed in the normal urothelium of "secretors" versus "nonsector" individuals. The normal urothelium of "secretors" is rich ABH, Leb, and Ley antigens while the urothelium of "nonsecretors" does not express these antigens. Therefore, deletion of ABH antigens, commonly noted in TCC, can only be reliably ascertained in "secretor" individuals. Neoexpression of the Lewis X antigen (which is absent in normal urothelium) is noted in over 85% of TCC regardless of tumor stage and grade. Immunocytological detection of the Lewis X antigen on exfoliated bladder epithelial cells enhances the detection of urothelial tumor cells, particularly from low grade and low stage neoplasms.


Chemotherapy for urothelial tract malignancies: breaking the deadlock. Chemotherapy treatments for urothelial tract tumors have improved to the point that some patients are enjoying long-term disease-free survival. Moreover, with currently available agents and combinations, and with our increased application of clinical and biologic prognostic factors, we are refining our ability to select appropriate therapies for individual patients. We have learned that once the decision is made to use combination chemotherapy, adequate doses should be used. This can be facilitated by the coadministration of hematopoeitic growth factors. Recently completed phase II trials have confirmed that higher doses and dose rates may increase response proportions of and in particular, the proportion of complete responses. The finding that granulocyte colony stimulating factor enhances the sensitivity of tumor cells to methotrexate in vitro and to other agents studied against urothelial tumors implanted in nude mice implies an expanded role for these compounds. However, because non-hematologic toxicities are still important, it is unlikely that simple escalation of all components a four drug regimen such as of M-VAC (cisplatin, methotrexate, vinblastine, and doxorubicin) will have a significant impact on survival. In addition, as more is learned about the pharmacokinetic and pharmacodynamic relationships of the active agents, it appears that better schedules can be designed to improve the therapeutic index of the compounds. Ultimately we will be able to determine drug sensitivities, both at the start of therapy and as it evolves during treatment, that will allow a better selection of a particular chemotherapeutic regimen. For example, mdr1 induction appears to play a significant role in the therapy for treatment-resistant tumors. The availability of a number of active salvage regimens that are not constrained by this mechanism hints that changes in drug sequencing and drug scheduling may provide a significant improvement in outcome. While established combination chemotherapy regimens should be considered standard therapy in appropriately selected patients, promising strategies and new agents need to be investigated if we are to "break the deadlock" that has appeared in the treatment of urothelial tumors. These investigations can be performed safely in a well-controlled fashion to enable the identification of new regimens and to compare promising strategies with appropriate control populations in randomized trials.


[Quantification of number and function of mononuclear cells. Fc gamma receptors in children with protein-calorie malnutrition (PCM)]. Several kind of cells T membrane receptors have been recognized, between them immunoglobulins heavy chains receptors mu and gamma, they act in the antigen recognition, they are subpopulations-cells T surface-matched too. In the malnutrition protein-calorie (MPC) immunologic alterations were observed, in both cellular immune and humoral immune, for example: The immunoglobulins are increased and cellular response towards antigens is decreased. This failure can be due to dysfunction in the lymphocyte T-subpopulation. So we research if there are different number, percent and function of lymphocytes T subpopulations between children with MPC and health infant. We observed an increment of lymphocytes T gamma in number and percent in children with MPC compared with health infant (p < 0.05) However, the lymphocytes T and subpopulation gamma response to phytohemagglutinin was bad. These findings are concordant with the literature about increased in lymphocyte T gamma of children with MPC. Additionally we observed a dysfunction in this subpopulation.


[Immunoglobulin A and tobacco antigens in patients with pneumopathy]. Smoking is an addiction related with several cardiopulmonary diseases. Some compounds derived from tobacco combustion can be induce a response of secretory immune system, because antigen penetration is by respiratory tract epithelium, with IgA antibodies synthesis and immune complex (IC) generation. Seric anti-tobacco antibodies were showed in 44% of healthy smokers and 71% of non-smoker. In 56% of smokers and 38% at non smokers were found IC with 0.19 and 0.15 mg/ml of IgA respectively. Molecular weight of IC constituents were between 12 and 80 Kd. Sera were without free tobacco antigen. Pneumopathic patients had anti-tobacco antibodies in 100% of them and positive IC in 72% of smokers and 65% of non-smokers. IgA concentration in CI was 1.41 and 1.26 mg/ml respectively. Molecular weight of IC compounds were from 14 to 90 Kd. Free tobacco antigen in serum was observed in 44% of smokers and 41% of non-smokers pneumopathic patients. We concluded that patients with lung disease had higher frequency of anti-tobacco antibodies and IC, further IgA concentration in IC was higher and free tobacco antigen present in pneumopathic sera comparing with healthy people. The presence of circulating IC with IgA antibodies and the potential inductive influence of tobacco products in these entities, should provide an energetic stimulus to search the role of tobacco antigens in pulmonary inflammatory diseases.


[Symptomatic dermatographism and HLA antigens]. This research reports the unique case of manifestation of symptomatic dermographism which includes all of its twenty five members of the family. Probing a genetic transmission autosomal dominant manner and taking into account the highly probable role of HLA in this illness. The symptomatic dermographism correspond to a group of physical familiar urticarias with symptoms appearing in early childhood and evolving throughout adulthood with seldom periods of remission. Due to the fact that all patients were affected, it was not possible to realize a typification of HLA antigens with persons not affected, to establish a relationship with the data obtained. However, results show a significant association of haplotypes; HLA A2 (78.95), B16 (68.4%), A1 (47.3%), and B5 (42.5%) among the most important. This is the first research that reports the analysis of HLA within the physical familial urticarias.


[Sensitization to the acarus Blomia tropicalis in patients with respiratory allergy]. House dust mites are important environmental allergens in Southern Brazil. Some allergic patients do not respond to specific immunotherapy for Dermatophagoides ssp. For that reason we decided to evaluate the skin test sensitivity to tropical mites, Blomia tropicalis and Blomia kulagini. Patients with perennial allergic rhinitis and/or asthma were selected for tests. (N = 135). The wheals cross diameter were recorded. The frequency of strong reactions were 82% for B. tropicalis, 69% for D. pteronyssinus, 56% for D. farinae and 46% for B. kulagini. Wheals were larger with Dermatophagoides sp in 30 patients (22%); similar in 61 patients (45%) and larger with-Blomia sp in 44 patients (33%). Three patients reacted only to Blomia. This study verifies that sensitization to Blomia is common in patients with respiratory allergy. In addition it contains unique allergens that could account for treatment failures with Dermatophagoides in some cases.


[3 years of "new developments" in pharmacotherapy. An update which is also a proposal for participative experimentation]. The analysis of the contents of key national and international drug bulletins provides a comprehensive overview of what has been produced worldwide in the field of pharmacology and therapeutics over the last few years after the last publication in R d 1). The work presented, tries to avoid the risk of being a purely descriptive listing of true and false "new" information, and may be seen as an "experiment of information transfer": the available data are presented and summarised from various complementary points of view, to suggest also a methodology of permanent education, for which instruments are also proposed. Further, the text is tentatively proposed as an experiment of communication between nurses and pharmacists, to stimulate their dialogue and interaction both with respect to the use of drug literature and to the evaluation of daily needs of information in routine hospital practice.


[The methods and instruments of nursing research: an analysis of some articles]. Three research papers published in three of the most respected foreign nursing journals are presented and discussed. The main aim of the contribution, which opens a new arena for discussion in the Rivista dell'Infermiere is to critically appraise published research works focusing on their strengths and weaknesses in the hypothesis formulation, methods and instruments used, discussion of results. A critical analysis should enable nurses to start to learn to read and eventually write a research protocol, possibly avoiding some common mistakes.


[Nursing diagnoses].

[Nursing diagnoses]. The history and the evolution of nursing diagnoses in the USA are briefly presented, together with the main controversies and objections against their widespread introduction and use in Europe. Though the creation of a nursing diagnoses taxonomy might be a very exacting but culturally stimulating task, one of the main problems to overcome is the identification of a theoretical framework. Clinical and theoretical research is needed to validate in our culture the process of formulation and the practical implications of nursing diagnoses identified in the States.


[A window on the past: nurses and education]. The history and the state of the art of the debate on nursing education in the first two decades of 20th century is presented. Improvement of nursing education was a strongly felt need from nurses and physicians: feminist movements were strongly in favour of a limited role of physicians in nursing education. Nurses trade unions (Leghe degli infermieri) supported an improvement of nurses' education as a mean to protect patients rights for a better care. Only a minority of physicians opposed the introduction of the English model of nursing care, with the matron who had the power of organizing the hospital. After the advent of fascism the law of 1929 on institution of Scuole Convitto was approved, and this law hindered the development of a reform of nursing education up to the Seventies.


Anergy and suppression in B-cell responses. Two main ideas have been put forward to explain the unexpectedly low anti-hapten antibody titres which can result from pre-priming a mouse with carrier before hapten-carrier immunization. The first involves the interaction of a network of idiotype-specific suppressor T cells, the second instead arguing for the role of intrinsic B-cell anergy. This paper proposes that the data available can equally be interpreted as reflecting the suboptimal interaction between T and B cells at differing stages of maturity, provided that memory B cells can be divided into two subsets. Further, it is suggested that these considerations must be taken into account in the analysis of B-cell anergy in receptor transgenic mice.


Anti-Golgi antibody in rheumatoid arthritis patients recognizes a novel antigen of 79 kDa (doublet) by western blot. We have detected cytoplasmic anti-Golgi antibody (AGA) during a routine immunofluorescence test for detecting autoantibodies. Two sera from patients with rheumatoid arthritis (RA) reacted to the Golgi complex by an indirect immunofluorescence technique on HEp-2 cells. Localization of AGA in the Golgi complex was confirmed by double-staining with antibodies to beta-COP. The effect of monensin on the integrity and morphology of the Golgi complex was also studied. To confirm the presence of AGA further, we performed immuno-electron microscopy. Both sera reacted with cytoplasmic antigen located in the Golgi complex of various animal tissues. Furthermore, by using the Western blot technique, both sera reacted to a relative molecular weight (MW) of 79 kDa (doublet) Golgi antigen purified from rat liver. To our knowledge, this study may be the first to identify the relative MW of Golgi antigen by the Western blot method. Identification of this antibody could provide better understanding of protein synthesis and secretion. The presence of AGA in RA patients further substantiates the diversified nature of autoantibody production seen in this disease.


Studies of antibody to herpes simplex virus Fc gamma-binding protein gE in patients with rheumatoid arthritis, juvenile rheumatoid arthritis and normal controls. IgG antibody to gE, the Fc gamma-binding herpes simplex 1 (HSV-1) viral glycoprotein, was studied in 49 rheumatoid arthritis (RA) patients and 43 normal controls. Antibody to gD, another important HSV-1 antigen, was assayed in parallel. No difference between RA patients and normal controls was found in levels of anti-gE antibody measured by reactivity of IgG F(ab')2 fragments reacting with gE coated to ELISA plates. No difference in anti-gD antibody was recorded between normals and patients with RA. Levels of IgG anti-IgE antibody did not correlate with quantitative elevations of serum rheumatoid factor (RF) in RA patients. When IgG anti-gE and anti-gD were assayed in 20 patients with juvenile rheumatoid arthritis and 22 children controls, no significant differences were noted. However, when individual RFs from patients with RA were tested for reactivity against a panel of affinity-isolated F(ab')2 antibodies to gE, some evidence for individual autospecificity was obtained. Four of 20 monoclonal IgM RFs produced from RA patients' B cells showed marked elevations of reactivity with some RA patients' F(ab')2 antibodies to gE. All four of the monoclonal RFs showing this specificity were derived from RA synovial tissue B cells. These findings may provide support for the concept that some RFs in patients with RA show individual specificity for internal image determinants of IgG antibodies to viral Fc gamma-binding proteins.


Induction of cellular immune reactions by A36, an antigen complex of Mycobacterium paratuberculosis: comparison of A36 and johnin components. Paratuberculosis (Johne's disease) is a chronic enteritis syndrome of ruminants, which is due to infection by Mycobacterium paratuberculosis. Cutaneous testing with proteins extracted from a mycobacterial culture fluid (johnin-PPD) is currently used to evaluate the cellular immune status. We have compared the components of johnin-PPD with those of the A36 complex, a thermostable macromolecular antigen (TMA) present in the cytoplasm and associated with the cell wall of M. paratuberculosis. The presence in the johnin-PPD of fifteen A36 components has been shown by Western blotting. Moreover, monoclonal antibodies, which bind respectively to the 65-kDa M. leprae heat shock protein, the 28-kDa M. leprae superoxide dismutase, and M. tuberculosis lipoarabinomannan, recognized components of the johnin-PPD. The ability of A36 to trigger delayed hypersensitivity reactions in sensitized rabbits, and to induce the proliferation of T lymphocytes from the lymph nodes of A36-sensitized mice, matched that of johnin-PPD. The homology levels of T epitopes between A36 and the TMA complexes of M. phlei, M. bovis, M. tuberculosis and M. avium were estimated, in a lymphoproliferation assay, to be 51, 52, 59 and 94% respectively. A strong cross-reactivity of A36 with an M. leprae sonicate was also observed by cutaneous testing. The A36 components within the 45.2-26.8-kDa and the 21.6-19.8-kDa ranges were proved to induce the proliferation of T lymphocytes from sensitized mice. This work supports the possible use of the A36 complex, and of some of its components, for cutaneous tests and lymphocyte proliferation assays, in order to monitor cellular immunity in Johne's disease.


Identification of immunodominant antigens during infection with Mycobacterium tuberculosis. T lymphocytes isolated from mice infected with Mycobacterium tuberculosis response vigorously to proteins secreted by the bacilli and these antigens may be of importance in the generation of protective immunity against the disease. In this study, short-term culture filtrate (ST-CF), which constitutes a complex mixture of secreted proteins, was fractionated by a modified preparative SDS-PAGE technique. The ability of each fraction to be recognized by T cells isolated from infected mice was evaluated by quantifying proliferation and IFN-gamma production in cell cultures. Two molecular mass regions 4-11 and 26-35 kDa were found to possess marked stimulatory properties. Four potent single antigens were mapped within the stimulatory regions. These purified antigens stimulated T cells isolated from mice at the height of a tuberculous infection to produce large amounts of IFN-gamma. Two of these stimulatory antigens belonged to the antigen 85 complex.


Cytokine production and responsiveness of fetal T-cell receptor V gamma 3 thymocytes. The aim of this study was to examine the cytokine production and cytokine responsiveness of the first T-cell receptor (TcR) positive cells that appear in the murine fetal thymus, namely TcR V gamma 3 cells. It is shown that IL-2-cultured fetal TcR V gamma 3 thymocytes were capable of producing IL-3, GM-CSF, TNF-alpha and IFN-gamma upon TcR triggering. IL-2, IL-4, IL-5 and IL-6 could not be detected. With regard to cytokine responsiveness, TcR V gamma 3 cells proliferated to a high extent when high concentrations of rIL-2 were added. rIL-4 or rIL-7 alone, but not rIL-1 alone, were capable of inducing a modest proliferation of TcR V gamma 3 thymocytes. When combined with low concentrations of IL-2, a synergistic effect could be observed with IL-1, IL-4 or IL-7. It is shown that the synergistic effect of IL-2 with IL-4 was mainly due to induction of IL-2 receptor expression. The synergistic effect of IL-2 and IL-7 on the proliferation of TcR V gamma 3 cells could only be partially inhibited by anti-IL-2 receptor MoAb, and this antibody had no effect on the IL-2 + IL-1 cultures. These observations can explain the extensive proliferation of TcR V gamma 3 thymocytes during fetal life and they indicate that TcR V gamma 3 thymocytes have the potential to play a functional role during fetal thymus development.


Spontaneous in vivo gene transcription of interleukin-2, interleukin-3, interleukin-4, interleukin-6, interferon-gamma, interleukin-2 receptor (CD25) and proto-oncogene c-myc by rheumatoid synovial T lymphocytes. Rheumatoid synovial T lymphocytes were investigated for the presence of mRNA for the cytokines interleukin-2, -3, -4, -6, interferon-gamma, the interleukin-2 receptor (CD25) and the proto-oncogene c-myc. The isolated RNAs were analysed by dot blot and Northern blot hybridization. Our results show that synovial T lymphocytes from patients with rheumatoid arthritis (n = 12) had spontaneous in vivo gene transcription of interleukin-2 (93%), interleukin-4 (67%), interleukin-6 (92%), interleukin-2 receptor (92%) and the proto-oncogene c-myc (67%). Only a few of the RA patients had synovial T cells with increased expression of mRNA for interleukin-3 (25%) and interferon-gamma (25%). The amounts of mRNA for the various cytokines and activation molecules produced by the rheumatoid synovial T lymphocytes were in most instances comparable to those of normal peripheral blood T lymphocytes activated in vitro by the mitogen phytohaemagglutinin. The data thus indicate that the synovial T lymphocytes are activated in vivo in the majority of rheumatoid arthritis patients.


A sensitive simple ELISA for quantitation of sensitizing IgG from dissolved erythrocytes. A simple enzyme-linked immunosorbent assay has been developed for quantitating the rabbit IgG present on the surface of sensitized red blood cells. A suitable cell lysis was carried out by an alkaline buffer, which dissolved the erythrocytes without forming any precipitate and without disruption of IgG, and facilitated the dissociation of the immune complexes, i.e. the erythrocyte-anti-erythrocyte rabbit IgG. In this alkaline buffer of pH 11.4 the IgG adsorbed directly into wells of microtitration plates unprecoated with anti-rabbit IgG. The detection was performed with alkaline phosphatase-conjugated goat anti-rabbit IgG and p-nitrophenyl phosphate as substrate. Constructing a calibration curve from rabbit IgG made possible the calculation of molecules of IgG bound per red cell. The method was sensitive for the detection of fewer than 500 bound IgG molecules per erythrocyte.


Phagocytosis following translocation of the neutrophil b-cytochrome from the specific granule to the plasma membrane is associated with an increased leakage of reactive oxygen species. The effect of neutrophil b-cytochrome translocation on the respiratory burst activation generated during phagocytosis of yeast particles was investigated. Secretion of neutrophil specific granules was induced by the calcium ionophore ionomycin prior to phagocytosis. The secretory process is associated with a translocation from the specific granules to the plasma membrane of the respiratory burst b-cytochrome. Respiratory burst activity was measured as release of hydrogen peroxide in the absence of azide (extracellular leakage) and in the presence of azide (total production). The subcellular localization of the b-cytochrome was found to affect the extracellular release of hydrogen peroxide in that a plasma membrane localization was associated with a significantly increased release during phagocytosis. It should be pointed out, however, that most of the hydrogen peroxide, both in control and in ionomycin-treated cells, is produced intracellularly, probably in the phagosomes.


T-cell receptor identification of an oligodendrocyte-specific autoreactive cytotoxic T-cell clone without self restriction. In addition to myelin basic protein (MBP) and proteolipid protein (PLP), oligodendrocyte (Od) membrane autoantigens, such as the glycoprotein M2/MOG, could participate in the pathogenesis of autoimmune demyelinating diseases of the central nervous system (CNS), such as experimental allergic encephalomyelitis (EAE) or multiple sclerosis (MS). We have described an Od-specific autoreactive and cytotoxic T-cell clone, named C2, which recognized M2/MOG without conventional MHC restriction. In order to analyse the Od/C2 interaction, we determined the alpha/beta T-cell receptor (TCR) variable region usages and structures of C2. Monoclonal antibody stainings of C2 and nucleotide sequences show that the alpha chain is composed of a V alpha 5 and a J alpha identical to J alpha 18BBM142 gene segments, and that the TCR beta chain is composed of V beta 17a, D beta 2.1 and J beta 2.2 gene segments indicating that C2 used a conventional alpha/beta TCR for M2/MOG recognition.


Quantitative imaging ion microscopy: a short review. A short review of recent efforts being made in the quantification of images in ion microscopy is given. Special aspects of instrumentation, detection and acquisition, which are unique to direct imaging secondary ion mass spectrometry, are discussed in relation to the successful application of traditional empirical quantification schemes. Application of such quantification schemes requires proper sample preparation, standardization, analysis, and quite often, special techniques in image processing and the correlation of ion microscopy with other microscopies. Quantification within this technique is a difficult goal which can only be realized if the analyst pays strict attention to every step of the analytical process.


Scanning tunneling microscopy of biological macromolecular structures coated with a conducting film. We have studied the capability of scanning tunneling microscopy (STM) to reveal the three-dimensional structure of biological macromolecular structures that have been rendered conductive by metal-coating. The sample preparation used has been derived from a well established method in transmission electron microscopy (TEM). It includes adsorption, freezing and dehydration by vacuum-sublimation, followed by metal-shadowing of the specimen. As an alternative to adsorption and coating, fluid biomaterials can be replaced by conductive freeze-fracture replica. We give an introduction into the sample preparation of metal-coated specimens and discuss how each step can affect the structural preservation and thereby the quality of the data. Some aspects of the data acquisition and the quantitative evaluation of STM data are shown. Possible contributions of STM in the biological macromolecular research are pointed out.


Can we see living structure in a cell?

Can we see living structure in a cell? Colloid chemistry (kappa o lambda lambda alpha: glue, or gelatin) was introduced in 1861 after the discovery of protoplasm which exhibits gelatin-like properties. Some 80 years later, colloid chemistry (and with it, the concept of protoplasm) was largely abandoned. The membrane (pump) theory, according to which cell water and cell solute like K+ are free as in a dilute KCl solution, became dominant. Later studies revealed that rejecting the protoplasmic approach to cell physiology was not justified. Evidence against the membrane (pump) theory, on the other hand, has stood the test of time. In a new theory of the living cell called the association-induction (AI) hypothesis, the three major components of the living cell (water, proteins and K+) are closely associated; together they exist in a high-(negative)-energy-low entropy state called the living state. The bulk of cell water is adsorbed as polarized multilayers on some fully extended protein chains, and K+ is adsorbed singly on beta- and gamma-carboxyl groups carried on aspartic and glutamic residues of cell proteins. Extensive evidence in support of the AI hypothesis is reviewed. From an extension of the basic concepts of the AI hypothesis and the new knowledge on primary structure of the proteins, one begins to understand at long last what distinguishes gelatin from other proteins; in this new light, new definitions of protoplasm and of colloid chemistry have been introduced. With the return of the concept of protoplasm, living structure takes on renewed significance, linking cell anatomy to cell physiology. Finally, evidence is presented showing that electron microscopists have come close to seeing cell structure in its living state.


Correction for extraneous background in X-ray microanalysis of cell cultures. Some practical aspects of the X-ray microanalysis of cell cultures have been investigated. Cells were cultured on titanium grids covered with Formvar films and analyzed at 100 kV either in the scanning transmission (STEM) or transmission mode (TEM) of the electron microscope. Different holders, grids and configurations were compared with respect to the relative contribution of different factors to the extraneous background in the X-ray spectrum. When low atomic number holders are used, the contribution to the spectrum of electrons scattered through high angles, may be negligible. In practice this may result in negative values for the contribution of these scattered electrons to the background. Computer programs for correction of the extraneous background should ignore these negative values and replace them by zero. When a brass holder is used, the contribution to the spectrum from electrons scattered through high angles becomes more important than that of the uncollimated radiation. The position of the analyzed cell relative to the grid bars is more important than the choice of grid or holder type. The data show that for the specimens used in the present study the correction for extraneous background is of little importance and can be neglected.


An improved method for preparing microvascular corrosion casts of rat embryos. This paper presents an improved method for preparing microvascular corrosion casts of rat embryos (day 16.5-21.5). Special attention was paid on the viscosity of the casting material, the method of mechanical restraint, and the subsequent drying of the casts. The embryos were perfused with Mercox resin diluted with 25% methyl methacrylate monomer followed by undiluted Mercox resin to avoid outflow of resin from the vasculature after perfusion. The specimens were mounted on a stainless steel plate with self-curing resin to prevent flotation, mechanical damage, and collapse of the cast specimens following preparation. After digestion and washing, the cast specimens were freeze dried to prevent deformation of the casts. This specimen preparation resulted in much better scanning electron microscopic images of the embryonic microvasculature of various oral tissues, free from leakage and insufficient filling.


Scanning electron microscopic studies of the oral mucosa and its microvasculature: a review of the palatine mucosa and its microvascular architecture in mammals. The present paper deals with the microvascular architecture of the palatine mucosa in primates, carnivorae, and rodentia utilizing microvascular corrosion castings and epithelium-separated specimens. The submucous vascular network is under-developed since the hard palatine mucosa was designated the mucoperiosteum, except some areas. The palatine venous plexus appears to show regional differences with animal species differences. The well-developed plexus is observed to be two-layered and may contribute to the process of regurgitation of rough food and assist in mastication with the palatine plicae. Formation and patterns of the arterial network in the lamina propria are in a close relation with connective tissue elements. The subepithelial capillary network constitutes an advanced base for the ascending crus of the capillary loop and its pattern is affected by the properties of the connective tissue papillae and the diverging fashion of the capillary loops. Capillary loops of the transverse palatine plica are arranged parallel to the sagittal axis and at right angles to the top line of each plica. Features of the capillary loops are characteristics in the top, the anterior and posterior slopes of the plica, respectively. High connective tissue papillae in both the anterior slope and plical top may be of a resistant form for mollifying exhaustion, affected by the periodicity and mastication function. Although it is difficult to elucidate the lamination of the palatine mucosa in histological slides, it was resolved by examination on its vascular architectures.


Scanning electron microscopy and energy-dispersive X-ray microanalysis studies of early dental calculus on resin plates exposed to human oral cavities. Dental calculus formed after 10 days on resin plates, applied to the lingual sides of the mandibular gingival regions in eight human subjects, was investigated by means of scanning electron microscopy (SEM) and energy-dispersive X-ray microanalysis (EDX). The mineral deposits were mainly divided into three types: A, B, and C. The type A deposits showing an average Ca/P molar ratio of 1.42 were densely packed with fine needle-shaped crystals formed by the intra- and extra-cellular calcification. The type A deposits, probably composed of Ca-deficient apatites and the transitional forms between apatite and octacalcium phosphate (OCP), were observed in all subjects. The type B deposits showing an average Ca/P molar ratio of 0.96 were aggregated with polygonal column, triangular plate-shaped, and rhombohedral crystals. These crystals identified as brushite (CaHPO4-2H2O:dicalcium phosphate dihydrate: DCPD) were found in four subjects. Platelet-shaped crystals of the type C deposits were observed in three subjects. Their Ca/P molar ratio of 1.26 and the crystal shape were similar to those of OCP. Whitlockite crystals were not found although Mg-containing hexagonal disk-like crystals were observed in two subjects.


Scanning electron microscopy-energy dispersive spectroscopy and X-ray diffraction analyses of human salivary stones. Ten salivary stones in the human submandibular gland were investigated by scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) and X-ray diffraction analyses. The stones usually showed a lamellar pattern. SEM observations revealed cubical, plate-like, granular, small and large granules, polyhedral or globular structures in these stones. By X-ray powder diffraction analysis, the main constituents of salivary stones were found to be apatite and whitlockite. SEM-EDS analyses showed that Ca and P were the major elements, frequently accompanied by Mg and S, and less frequently by Na, Al, Si, Cl, K, Fe, Cu and Zn. Ca/P molar ratios ranged from 1.00 to 2.00 with the average of 1.53, showing two maxima of about 1.50 and 1.60. The Ca/P molar ratio of about 1.50 corresponded to the value of whitlockite. The Ca/P molar ratio of 1.60 corresponded approximately to the value of apatite.