The pathway of arginine catabolism in Giardia intestinalis. In Giardia intestinalis, arginine is catabolised by the arginine dihydrolase pathway. The enzymes of the pathway (arginine deiminase, ornithine transcarbamoylase and carbamate kinase) were investigated and their basic kinetic parameters determined. The specific activity of arginine deiminase was 270 +/- 23 nmol min-1 (mg protein)-1; ornithine transcarbamoylase, in the direction of citrulline utilisation 170 +/- 22 nmol min-1 (mg protein)-1, and in the direction of ornithine utilisation 2100 +/- 100 nmol min-1 (mg protein)-1; and carbamate kinase 2100 +/- 400 nmol min-1 (mg protein)-1. The activities of these enzymes are between 10 and 250 fold greater than those reported for the enzymes in Trichomonas vaginalis, the only other parasite in which the arginine dihydrolase pathway has been reported. The flux through the pathway in G. intestinalis, as determined by the liberation of 14CO2 from 1 mM [14C-guanidino]arginine was 30 nmol min-1 (mg protein)-1. This flux was not affected by valinomycin (0.1 microM), nigericin (3 microM), azide (5 mM) or cyanide (1 mM). The flux was only marginally affected by glucose up to 10 mM concentration. Conversely, the flux through glucose metabolism, as determined by the release of 14CO2 from 1 mM [1-14C]glucose was only 2 nmol min-1 (mg protein)-1, and was unaffected by arginine concentrations up to 10 mM. These observations suggest that there is no direct metabolic interface between arginine and glucose catabolism.(ABSTRACT TRUNCATED AT 250 WORDS)


Heparin, fatty acids and sodium, potassium-ATPase inhibition by plasma factors during hemodialysis. To assess the relationship between heparin and the associated increase in nonesterified fatty acids (NEFA) and their possible influence on Na,K-ATPase during hemodialysis, we studied two groups of patients: (1) 12 patients on chronic hemodialysis dialysed with heparin and (2) 6 patients dialysed without heparin. All 12 patients who received heparin anticoagulation had a 7-fold rise in NEFA on average and also had an increase in circulating inhibitors of Na,K-ATPase assayed by 3H-ouabain displacement from Na,K-ATPase and/or by effect of plasma on the uptake of 86Rb by rat aortic rings. Serial assays in 3 of the patients receiving heparin showed NEFA and inhibitory changes to be at or near maximum within 30-60 min. Of the individual NEFA, the greatest relative increases were in oleic (18:1) and linoleic (18:2) acids, and the strongest correlations were between linoleic acid and both 3H-ouabain displacement (r = 0.94) and 86Rb uptake (r = 0.86). However, a small and slower increase in NEFA also occurred in 3 of the patients dialysed without heparin. We conclude that heparin anticoagulation during dialysis leads to a rapid and marked increase in circulating NEFA, particularly the unsaturated fatty acids, with a corresponding interference with Na,K-ATPase activity. The clinical significance of these findings is unknown. The rise in NEFA during dialysis without heparin in some patients suggests that factors other than heparin may also contribute to the rise in NEFA.


Effects of methylguanidine on sodium and organic ion transport. We studied the acute effect of methylguanidine (MG), a suspected uremic toxin that accumulates in renal failure, on p-aminohippurate (PAH) and tetraethylammonium (TEA) uptake in rabbit kidney slices, on Na+,K+ ATPase activity in the microsomal fraction of rabbit kidneys, and on transepithelial active Na transport across toad skin. MG at concentrations ranging from 0.05 (similar to that reported in uremic patients) to 1.0 mM does not affect the organic anion (PAH) uptake, although it exhibits a concentration-dependent inhibition of organic cation (TEA) uptake. MG at concentrations from 0.05 to 5 mM had no effect on kidney Na+,K+ ATPase activity or on active transepithelial Na transport across toad skins when applied to the outside bathing solution; however, MG (greater than 1 mM) stimulated Na transport when applied to the inside bathing solution. These results are not consistent with the hypothesis that MG is a potential uremic toxin that causes the natriuresis and other toxic effects. However, long-term toxic effects of MG on the kidney were not assessed in the present study.


Neurohypophyseal and pituitary-adrenocortical responses to the alpha 1 agonist methoxamine in humans. To test the hypothesis that the release of neurohypophyseal peptides into plasma in humans is stimulated by a central nervous system (CNS) alpha 1 adrenergic mechanism, we measured the responses of arginine vasopressin (AVP) and oxytocin (OT) to intravenous methoxamine, an alpha 1 agonist which enters the CNS following peripheral administration. The potential confound of baroreceptor inhibition of AVP release by the pressor effect of methoxamine was addressed by measuring the plasma AVP response to infusion of norepinephrine (NE), an alpha 1 agonist which does not enter the CNS and which produced an equivalent pressor effect. We also assessed the pituitary adrenocortical system responses to methoxamine and norepinephrine infusions by measuring plasma ACTH and cortisol concentrations. In addition, plasma NE and epinephrine were measured. Methoxamine, but not NE, increased plasma AVP compared to placebo infusion. Neither methoxamine nor NE affected plasma OT. The AVP elevation was delayed until more than 60 min after the methoxamine infusion began and the peak AVP level occurred 30 min after cessation of the infusion. In contrast, ACTH and cortisol increased early during methoxamine infusion and ACTH returned to baseline promptly after the infusion ceased. Although it is possible that the AVP response to methoxamine reflected stimulation of AVP release at a CNS level, it is also possible that the AVP increase represented a rebound response to withdrawal of methoxamine.


Restraint inhibits luteinizing hormone and testosterone secretion in intact male rhesus macaques: effects of concurrent naloxone administration. The present study investigates how restraint affects the hypothalamo-hypophysial adreno-cortical axis and the hypothalamo-hypophysial gonadal axis in intact, adult male rhesus macaques. Restraint was chosen because it is not physically painful or harmful to the animal, but rather serves both as a physical and psychological stressor. Blood samples were collected from a remote site at 15-min intervals beginning at 07.00 h from tethered adult male rhesus macaques. Each of 4 animals was subjected to 6 h of chair restraint after a 3-hour control period in the animals' home cage. Samples were collected for an additional 6 h at the end of the restraint period when the animal was returned to its home cage. Brief anesthesia with ketamine (administered through the indwelling catheter) facilitated transfer of the animals to and from the chair. Blood samples were collected from 4 undisturbed males to document LH and testosterone secretion throughout the day. Plasma ACTH and cortisol, measured as indexes of stress, were elevated within 15 min after initiation of restraint and remained elevated for most of the restraint period. Conversely, LH and testosterone began to fall immediately after restraint and remained suppressed for several hours after the animals were removed from restraint and returned to their home cage. Testosterone levels were more consistently inhibited than were LH levels, a reflection of the fact that in some animals, testosterone remained low after the return of pulsatile LH secretion. In studies with naloxone (Nx), the opiate receptor antagonist (5 mg bolus plus 5 mg/h) was given beginning either at the initiation of restraint (n = 2) or 2 h thereafter (n = 2), and continued until the end of the restraint period. With Nx treatment of the restrained animals, both ACTH and cortisol were elevated as in the controls and LH and testosterone secretion were significantly increased within 1-2 h. However, after the Nx treatment was terminated and the animals were returned to their home cages, plasma levels of LH and testosterone were not different from levels in restrained animals and were significantly less than levels in untreated animals. These data show that restraint is a potent stimulus for activation of the HPAC axis and inhibits both LH and testosterone release. The pathway through which restraint inhibits LH release probably includes endogenous opiate suppression of hypothalamic GnRH release since Nx partially blocks the effect of stress.


Estrogens regulate angiotensin-converting enzyme and angiotensin receptors in female rat anterior pituitary. We studied the effects of the estrous cycle, ovariectomy and estrogen replacement on angiotensin-converting enzyme (ACE) (kininase II, EC 3.4.15.1) and angiotensin II (AT) receptors in the pituitary gland of the female rat. Quantitative autoradiography, with the use of consecutive pituitary sections, allowed for simultaneous determination of changes in binding and in the potential AT synthetic ability of individual pituitaries, and for a correlation between these two phenomena. In the anterior pituitary, ACE activity and binding of the ACE inhibitor [125I]-351A were not changed during the estrous cycle. Ovariectomy produced a significant increase in ACE activity and binding, and both of these parameters returned to normal after estrogen replacement. There were no changes in ACE activity or binding in the posterior pituitary during the estrous cycle or after ovariectomy or hormone replacement. AT receptors were characterized as of the AT1 type, since they were displaced by the selective AT1 antagonist DuP 753 and not by the AT2 competitor PD 123177. There were marked changes in the concentration of AT1 receptors during the estrous cycle, with highest numbers in metestrus, lower in estrus and diestrus, and lowest during proestrus. Estrogen replacement in ovariectomized rats decreased AT1 receptor number in the anterior pituitary. Our results indicate a dual effect of estrogen on anterior pituitary AT, physiologically on AT receptor expression and pharmacologically on ACE activity.


Effects of adrenalectomy and glucocorticoids on rat brain dopamine receptors. The effects of adrenalectomy (ADX) and dexamethasone (DEX) treatment on brain dopamine (DA) receptors of ovariectomized (OVX) rats were investigated by autoradiography using binding of the D1 and D2 antagonists ligands [3H]SCH 23390 and [3H]spiperone, respectively. Fourteen days after ADX, D1 receptors decreased in the middle striatum (M.CPu) and in the dorsal area of the posterior striatum (P.CPu). A more pronounced decrease was observed in the substantia nigra (SN) and no significant changes occurred in the anterior striatum (A.CPu), globus pallidus (GP) and accumbens (Acb). D2 receptors decreased in the M.CPu and dorsal area of the P.CPu and remained unchanged in the A.CPu as compared to OVX rats. Twenty-eight days after ADX, D1 and D2 receptors decreased in the A.CPu (D2), in the M.CPu (D2) and substantially in the SN (D1). DEX treatment (14 days, 0.5 mg/kg, b.i.d., IM, starting 14 days after ADX) reversed these effects in the A.CPu (D2), M.CPu (D2) and SN (D1) when compared to ADX+OVX rats. DEX also increased the density of D1 receptors in Acb, A.CPu and M.CPu when compared to OVX rats. Striatal homogenates of rats treated chronically with ACTH and corticosterone had an increased density of D1 receptors while these treatments alone left these receptors unchanged, thus suggesting either a minor role or no role of the changes in ACTH levels following glucocorticoids manipulations. Our results suggest that the adrenals play a role in the modulation of DA receptors activity in the rat brain.


Alternative excitotoxic hypotheses.

Alternative excitotoxic hypotheses. The concept of excitotoxicity, neuronal death produced by overstimulation of excitatory amino acid receptors, has become a popular way of explaining the pathogenesis of neuronal death in a variety of acute and chronic neurologic diseases. While there is strong evidence supporting the role of excitotoxicity in acute processes such as hypoxia/ischemia and hypoglycemia, the role of excitotoxicity in chronic neurologic disease is not firmly established. To account for the inter- and intraregional variations in pathology of different neurodegenerative disorders, we suggest two modified forms of the excitotoxic hypothesis in which specific populations of neurons become more vulnerable to excitotoxic insult either by (1) possessing abnormal excitatory amino acid receptor subtypes or (2) being afflicted by any disease process that impairs cellular energy metabolism or otherwise decreases neuronal membrane potential. In these ways, excitotoxicity may be a final common pathway of neuronal death in a variety of neurodegenerative diseases.


"Central" and "peripheral" benzodiazepine receptors: opposite changes in human epileptogenic tissue. We measured the density of two benzodiazepine (BZ) receptor subtypes in neurosurgically obtained hippocampal tissue from the seizure focus of patients with temporal lobe epilepsy (TLE) showing mesial temporal sclerosis, the most common pathologic finding in TLE. We performed quantitative in vitro receptor autoradiography with [125I]Ro 16-0154, a probe for the central-type BZ receptor and with [3H]PK 11195, a probe for the peripheral-type BZ receptor. In comparison with autopsy and neurosurgical control groups, patients with mesial temporal sclerosis had regionally selective decreased central-type and increased peripheral-type BZ receptors. These changes paralleled regional losses of neurons and proliferation of glia. Decreases of the inhibitory central-type BZ receptor may be a component of the enhanced excitability of the seizure focus and also may allow localization of the focus by in vivo neuroreceptor imaging. Single photon emission computed tomography (SPECT) imaging of two TLE patients with [123I]Ro 16-0154 suggests that this technique may provide a more sensitive means of localizing the seizure focus than current imaging methods relying on changes in blood flow or glucose metabolism.


Trends in the association of Lambert-Eaton myasthenic syndrome with carcinoma. The Lambert-Eaton myasthenic syndrome (LEMS) is often associated with carcinoma. The exact number of patients with tumor has been reported to be as high as 70%. Recent clinical experience suggests that the actual number of patients with tumor may be substantially lower. We combined data from the clinical experience of the neuromuscular services at West Virginia University and the University of Virginia to determine the rate of occurrence of cancer in this disorder. We identified 28 patients with LEMS, and 14 had cancer. There is a distinct trend for a lower tumor frequency over the past decade, which suggests that the clinical manifestations of the disease may be changing.


Familial inclusion body myositis: evidence for autosomal dominant inheritance. We report a kindred manifesting clinical features and muscle biopsy findings of inclusion body myositis (IBM). In this family, multiple members were affected in two generations with direct male-to-male and female-to-male transmission. This is the first reported instance of autosomal dominant inheritance in IBM, which usually occurs sporadically or, rarely, may be transmitted as an autosomal recessive disorder.


Effects of hypoglycemia on the cerebral circulation in awake goats. We electromagnetically measured blood flow to one cerebral hemisphere and determined cerebrovascular reactivity to vasoconstrictor and vasodilator stimuli during normoglycemia and insulin-induced hypoglycemia in unanesthetized goats. Control blood glucose concentration was 84 +/- 4 mg, and insulin, injected intravenously, decreased glycemia with a concomitant increment in cerebral blood flow and reduction in cerebrovascular resistance in all the animals. When glycemia decreased to 60 to 65 mg/dl, the animals began to show signs of increased adrenergic activity, and when it decreased to less than 30 mg/dl, they showed signs of CNS depression. Cerebral blood flow began to rise significantly at a glycemia of 50 to 55 mg/dl, and progressively increased to reach an increment of 36% +/- 4% when glycemia was less than 30 mg/dl. Norepinephrine (0.3 to 9 micrograms), tyramine (50 to 500 micrograms), and 5-hydroxytryptamine (0.1 to 9 micrograms) reduced cerebral blood flow, and this effect was lower during severe hypoglycemia. Acetylcholine (0.01 to 1 microgram), isoproterenol (0.03 to 3 micrograms), diazoxide (0.3 to 9 mg), and inhalation of 10% CO2 in air increased cerebral blood flow, and this effect was also lower during severe hypoglycemia. The results show that insulin-induced hypoglycemia causes cerebral vasodilation and reduction of the capacity of cerebral blood vessels to constrict and dilate. They also show that the glycemic thresholds for increasing cerebral blood flow are near to, or slightly lower than, the thresholds for hypoglycemic symptoms. This experimental model of hypoglycemia closely resembles the conditions in hypoglycemic patients and permits serial evaluation of the cerebrovascular effects of hypoglycemia without using anesthesia.


[VIPoma: surgical treatment].

[VIPoma: surgical treatment]. This paper reports a case of pancreatic VIPoma with widespread hepatic metastasis which was treated for approximately 2 years with a synthetic somatostatin analog (SMS 201/995). The treatment of choice in cases in which the tumour was fully removable is surgical resection. This occurred rarely since approximately 80% of VIPomas are malignant and are operated late when local infiltration is already widespread; in addition, 50% of cases are already metastasised at diagnosis. In this case, due to the infiltration of the superior mesenteric artery by the primary tumour it was necessary to carry out a left pancreasectomy which included two-thirds of the neoplastic mass. This was justified by slow tumour growth and also facilitated control of diarrhea and ensured a greater efficacy of possible postoperative chemotherapy. The use of synthetic somatostatin analog (SMS 201/995) enabled diarrhea to be satisfactorily controlled and is therefore specifically indicated for this type of tumour. NSE serum assay (neuron specific enolase) allowed the evolution of disease to be monitored during follow-up.


Primary intra-abdominal malignant fibrous histiocytoma. Case report and literature review. Malignant fibrous histiocytoma is a pleomorphic soft tissues sarcoma usually found in the extremity and in the retroperitoneum. The primary intra-abdominal localization of this tumour is very rare and only thirty-five previous reports are present in the world medical literature. The Authors report a case of giant malignant fibrous histiocytoma of the mesentery, which is treated by surgical removal plus adjuvant chemotherapy.


[Vulvar papillary hidrocystadenoma. Description of a case]. Papillary hidradenoma of the vulva is a benign neoplasm arising from apocrine sweat glands of the skin. The treatment of choice is local excision. The prognosis for the patient is excellent. We present one case observed recently.


Adenosine and its nucleotides stimulate proliferation of chick astrocytes and human astrocytoma cells. Aqueous extracts of the brains of 18-day-old white Leghorn chicken embryos contain several substances that stimulate proliferation of primary cultures of chick brain astrocytes. Most of the mitogens are peptides. Purification of one mitogenic fraction was obtained by centrifugation, passage through Amicon Diaflo membranes of nominal molecular weight cutoffs 30, 1 and 0.5 kDa, ion exchange chromatography and reverse phase high performance liquid chromatography (HPLC) using a Deltapak C18 column. The mitogenic fraction contained no amino acids. On the basis of its behaviour on thin layer chromatography, its ultraviolet absorption spectrum, its 1H and 31P nuclear magnetic resonance spectra and its behaviour on positive and negative ion fast atom bombardment mass spectrometry, the mitogenic material was identified as adenosine-5'-monophosphate (AMP). Other adenosine compounds including adenosine, ADP and ATP also stimulated proliferation of and [3H]leucine incorporation into primary cultures of astrocytes. Nitrobenzylthyioinosine (NBTI), an inhibitor of nucleoside transport, did not prevent the stimulation of [3H]leucine incorporation into cultured astrocytes. Polyadenylic acid (Poly A), that mimics the effect of adenosine at adenosine receptors, also stimulated proliferation of the astrocytes. The effects of adenosine and Poly A were not inhibited by 1,3-dipropyl-8-(2-amino-4-chlorophenyl)xanthine (PACPX) but were inhibited by 1,3-dipropyl-7-methyl-xanthine (DPMX), indicating that adenosine and Poly A acted at the cell surface, likely through adenosine A2 receptors. The stimulatory effect of ATP was biphasic. The proliferative effect of low, but not of high, concentrations of ATP were abolished by DPMX. The purinergic P2 receptor agonist 2-methylthioATP and, at higher concentrations, the P2y agonist, alpha,beta-methyleneATP also stimulated incorporation of [3H]thymidine. These data indicate that high concentrations of ATP stimulate cell proliferation through at a P2, possibly a P2y receptor. These results have considerable biological significance. After brain injury, or when cells in brain die or become hypoxic, nucleotides and nucleosides are released from the cells. Their extracellular concentrations can exceed those required to stimulate astrocyte proliferation in vitro. Therefore they may be partly responsible for gliotic changes following cell death in brain.


Fractal dimension of dendritic tree of cerebellar Purkinje cell during onto- and phylogenetic development. The cerebellar Purkinje cell has highly branched dendrites extending in a transverse plane to the cerebellar folium. Branching patterns of the dendrites were subjected to fractal analysis to obtain a quantitative measure of their intricacy. Using a square-covering method, we found that dendrites have a statistical self-similarity so that their fractal dimension can be evaluated. In postnatal mouse, dimensions of Golgi-stained Purkinje cells increased as dendrites developed. The time course of the dimension increase was correlated with development of the width and area of the dendritic fields. After the dimension reached the maximum value at postnatal day 20, the width and area further continued to grow until day 100. In human cerebellum, dimensions of Purkinje cell dendrites increased greatly during pregnancy and reached 97% of the adult value at birth. On the other hand, 41% of the adult dendritic field width and 11% of the adult area had been attained at birth. The physical size of dendrites increased in a more lagged phase than the dimensions, and growth proceeded mainly after birth. These observations indicate that after the maximum complexity and fundamental tree pattern have been attained, the overall tree size further increases. It is observed that phylogenetic development of dendritic trees of Purkinje cells in several species of vertebrates showed a correlation with the increase in dimensions. These studies demonstrated the potency of the fractal analysis in evaluating development of dendritic trees of neurons.


Changes of drebrin expression in the visual cortex of the cat during development. The expression of and developmental changes in drebrin were studied in cat visual cortex using immunohistochemistry and immunoblot analysis. Drebrin is a developmentally regulated brain protein which in the chicken has characteristic changes in expression related to developmental stage. A monoclonal antibody (MAb M2F6) raised against drebrin, was found to label the neuropil of the kitten visual cortex in the early postnatal period. At 1-3 weeks of age, the staining was prominent in layer IV of the visual cortex. The immunoreactivity, however, was found to be dramatically decreased around the end of the sensitive period for ocular dominance plasticity (approximately 3 months of age). In the adult visual cortex, almost no immunostaining was observed. These developmental changes revealed by an immunohistochemical method were confirmed using immunoblot analysis. Upon immunoblot analysis after SDS-PAGE of protein from the kitten visual cortex, MAb M2F6 was found to recognize two protein bands with molecular weights of 130 kDa (drebrin E) and 140 kDa (drebrin A). The developmental profile of the intensity of the two bands of the drebin closely parallels in time the postnatal changes in cortical susceptibility to visual deprivation. These results indicate that the expression of drebrin in kitten visual cortex is restricted to the early postnatal period and suggest that it may play an important role in the experience-dependent modification of cortical circuitry during the sensitive period.


Cat 'P300' and cholinergic septohippocampal neurons: depth recordings, lesions, and choline acetyltransferase immunohistochemistry. The role of septohippocampal circuits in the generation of the P300 response in cats (n = 12) was explored in a series of depth recording, tract-tracing and lesion experiments. Systematic mapping of the hippocampus in 1-mm increments from rostral to caudal extent revealed large positive potentials, greater in amplitude to rare than to frequent stimuli, within the 200-500 ms range. Each map revealed maximal amplitude responses at diverse, widely distributed hippocampus loci. Furthermore, these electrical responses displayed polarity inversion within the hippocampus that was generally localized to the pyramidal cell layer; polarity inversion was also observed in the adjacent entorhinal cortex and amygdala. Injections of propidium iodide, a tract-tracing agent, into these inversion sites resulted in retrograde labeling of small clusters of choline acetyltransferase (ChAT)-positive neurons in the medial septal nucleus and vertical limb of the diagonal band. Aspiration lesions that bilaterally destroyed large amounts of caudal hippocampus from stereotaxic levels A4 to A1 resulted in a decreased number of cells expressing ChAT in the rostral basal nuclear complex. In only 2 cats was the preoperative presence of a significant vertex P300 absent postoperatively. In the majority of cases (5 of 8 animals), hippocampal aspiration produced an enhancement of the preoperative P300 potential. We conclude that cholinergic mechanisms are importantly, albeit not exclusively, involved in the mediation of P300 potentials in cats. Neurons mediating P300 responses appear to be organized in diverse clusters of septal and diagonal band cells. These septal cells may facilitate, and in turn be facilitated or inhibited as a function of hippocampal, or other, allocortical feedback loops.


Bandeiraea simplicifolia lectin I and Vicia villosa agglutinin bind specifically to the vomeronasal axons in the accessory olfactory bulb of the rat. The binding of 21 lectins to the accessory olfactory bulb (AOB) of the rat was examined by histochemistry. Two lectins [Bandeiraea simplicifolia lectin I (BSL-I) and Vicia villosa agglutinin (VVA)] bound specifically to the vomeronasal (VN) axons in the AOB. Seven lectins (Datura stramonium lectin, Erythrina cristagalli lectin, Lycoperisicon esculentum lectin, Ricinus communis agglutinin I, soybean agglutinin, Solanum tuberosum lectin, and Ulex europaeus agglutinin) bound to both VN axons in AOB and olfactory axons in the main olfactory bulb. BSL-I and VVA are useful as the marker of VN axons. This selective binding of lectins indicates the presence of specific glycoconjugates on the surface of VN axons.


Human papillomavirus prevalence in pregnancy. Conventional wisdom has long held that human papillomavirus (HPV) prevalence is increased in pregnancy. We examined cervical swabs of 375 women for HPV DNA using polymerase chain reaction (PCR) and ViraPap with an expanded probe range. Of this population, 115 were pregnant (42 in the first trimester, 46 in the second, and 27 in the third) and 100 were postpartum. The control population consisted of 160 women who were otherwise similar to the pregnant population but were not pregnant or postpartum. Crude associations were examined between HPV prevalence at defined high, low, and overall levels and the pregnancy status. Multivariate analysis indicated no statistically significant association between the prevalence at any level of infection and pregnancy status. As expected, associations were found between measures of HPV prevalence and both Papanicolaou smear results and warts seen on examination. No association was found for race, smoking behavior, or number of sexual partners and HPV prevalence. The estimated risk of an HPV infection decreased as the age of the women increased. We conclude that a significant relationship between pregnancy and HPV prevalence has yet to be established.


Natural history of cervical human papillomavirus lesions does not substantiate the biologic relevance of the Bethesda System. The newly introduced Bethesda System proposes to replace the terms dysplasia, carcinoma in situ (CIS), or cervical intraepithelial neoplasia (CIN) with two terms, low-grade squamous intraepithelial lesion (SIL) or high-grade SIL. We tested the biologic relevance of the Bethesda System (ie, its ability to establish a close correlation between biologic behavior and the different grades of lesions) in a series of 528 women with genital human papillomavirus (HPV)-associated precancerous lesions prospectively followed for 10 years. The cervical biopsies were reclassified as either low-grade or high-grade SIL, and the data obtained by colposcopy, Papanicolaou smear, and HPV typing, as well as the biological behavior of the lesions, were analyzed in these two groups. Altogether, 77.4% (376 of 486) of the lesions were classified as low-grade SIL lesions; the rest (22.6%) belonged to the high-grade SIL category. In the low-grade SIL category, 46.8% of the women were 24 years old or younger, as compared with 37.3% in the high-grade SIL group. The colposcopic appearance was normal significantly more frequently in the low-grade SIL lesions (22.1%) than in the high-grade category (8.5%) (P less than .001). A single Papanicolaou smear was inadequate to distinguish between low-grade and high-grade SIL, as evidenced by almost identical distributions of Papanicolaou smear class I and II cells in both categories. Noteworthy was the discovery of normal Papanicolaou smears in 8.2% of the high-grade SIL lesions.(ABSTRACT TRUNCATED AT 250 WORDS)


Repeated evaluation of human papillomavirus 16 status in cervical swabs of young women with a history of normal Papanicolaou smears. This study was designed to compare single-point prevalence estimates with a cumulative prevalence estimate of human papillomavirus (HPV) type 16 in cervical smears. The influence of the menstrual cycle and the long-term effect of HPV 16 positivity on the development of cervical intraepithelial neoplasia (CIN) were monitored. We examined 21 women (median age 23.6 years) every 5 weeks for 1 year. All women had a history of negative Papanicolaou smears for at least 5 years before enrollment. Cervical swabs were collected at each visit for Papanicolaou smears and HPV 16 detection by the polymerase chain reaction. Five years after completion of the last visit, self-reported information regarding cervical neoplasia was obtained. Human papillomavirus type 16 point-prevalence estimates per visit varied between 14.3-33.3%. The cumulative prevalence was 66.7%; 14 women were positive at least once and seven women were continuously negative for HPV 16. Detection of HPV 16 was significantly higher in the luteal phase. Repeated positivity for HPV 16 by Southern blot and polymerase chain reaction preceded and accompanied CIN in one patient, whereas in the remaining patients, positivity for HPV 16 by polymerase chain reaction alone was not associated with CIN during a 5-year follow-up. Single-point measurements of HPV 16 in cervical smears by polymerase chain reaction are of limited value for assessment of an individual's HPV status. This should be kept in mind when HPV testing for screening programs is considered.


Erythrocytosis associated with a placental-site trophoblastic tumor. We present a case of placental-site trophoblastic tumor associated with erythrocytosis. This 42-year-old woman had persistent amenorrhea and low elevations of her hCG titer after term delivery of a healthy female infant. The woman was noted to have polycythemia of uncertain etiology and was treated with serial phlebotomy. Placental-site trophoblastic tumor was diagnosed and hysterectomy was performed, with subsequent resolution of the polycythemia. Although erythrocytosis has been reported with other gynecologic tumors, this is the first reported association with a placental-site trophoblastic tumor. A role has been suggested for placental lactogen in erythropoiesis during pregnancy based on previous animal studies. Diffuse positive staining for hPL is characteristic of placental-site trophoblastic tumors. We postulate that hPL may have played an ancillary role to erythropoietin in the erythrocytosis demonstrated in this case. Spider angiomata and splenomegaly are interesting clinical features previously described in association with placental-site trophoblastic tumors, and were demonstrated in this patient.


Spontaneous resolution of nonimmune hydrops fetalis secondary to human parvovirus B19 infection. Many instances of nonimmune hydrops fetalis ascribed to human parvovirus B19 have been reported. The leading proposed pathophysiologic mechanism of hydrops in affected fetuses is viral invasion of red blood cell progenitors, causing a profound reticulocytopenic fetal anemia. Although the natural history of fetal parvovirus infection remains to be elucidated fully, there have been recent reports of funipuncuture and intrauterine blood transfusions to diagnose and manage this problem. We report two pregnancies in which parvovirus-related hydrops fetalis was observed to resolve without intervention, followed by uncomplicated vaginal deliveries of healthy infants. These observations emphasize the need for further investigation before recommending routine fetal blood transfusion in affected cases.


Influence of coralline hydroxyapatite used as an ocular implant on the dose distribution of external beam photon radiation therapy. Coralline hydroxyapatite spheres are used as buried integrated ocular implants after enucleation or evisceration surgery. Because such implants are used after surgery for intraocular malignancy and because some patients may require postoperative radiation therapy for orbital tumor recurrence, the radiation attenuation characteristics of the implant are of interest. The authors evaluated the attenuation and scattering properties of coralline hydroxyapatite implants using a 4 MV photon beam and film dosimetry. Optical density analyses indicate that coralline hydroxyapatite implants have no significant influence on the attenuation or scattering properties of the photon beam. As such, there is no basis for concern that such implants might adversely affect external beam photon irradiation.


Surgical repair of rhegmatogenous retinal detachment in immunosuppressed patients with cytomegalovirus retinitis. The authors performed retinal reattachment surgery in 29 eyes of 24 patients with acquired immune deficiency syndrome virus with retinal detachment associated with cytomegalovirus (CMV) retinitis and documented the course of eight additional untreated eyes. Retinal detachment repair using vitrectomy, posterior hyaloid removal, and intraocular tamponade with silicone oil or SF-6 gas resulted in a total retinal reattachment rate of 76% and a macular attachment rate of 90% in one operation. The mean postoperative visual acuity (best corrected) was 20/60, but, in some patients, the visual acuity decreased because of progressive retinitis. Prophylactic laser photocoagulation of fellow eyes to surround CMV lesions did not appear to prevent retinal detachment. The mean postoperative survival was 37 weeks (range, 8 to 127 weeks). The surgical techniques used and pathophysiology of these retinal detachments are discussed.


Recombinant retroviruses encoding human papillomavirus type 18 E6 and E7 genes stimulate proliferation and delay differentiation of human keratinocytes early after infection. Human papillomavirus (HPV) DNAs are detected in most genital dysplasias and cancers, suggesting that these viruses perturb epithelial growth and differentiation. The E6 and E7 genes of HPV type 18 induce immortality in keratinocytes cultured from genital tract epithelia, and the immortal cell lines display aberrant squamous differentiation. To examine whether the E6 and E7 proteins directly alter keratinocyte growth and differentiation, high-titer recombinant retroviruses were constructed for efficient transfer and expression of HPV-18 genes E6, E6* and E7 in cultures of normal human keratinocytes. Infection with retroviruses encoding E6 and E7 stimulated cell proliferation, reduced the requirement for bovine pituitary extract and induced immortality. E6 and E7 also delayed but did not prevent the onset of terminal squamous differentiation. The magnitude of effects on growth and differentiation of cultured cells was directly related to levels of E7 protein expression. Thus, expression of the HPV-18 E6 and E7 genes stimulates cell proliferation and delays differentiation of keratinocytes in vitro.


Amplification of alpha-platelet-derived growth factor receptor gene lacking an exon coding for a portion of the extracellular region in a primary brain tumor of glial origin. In a primary brain tumor of glial origin, we found overexpression of the alpha-platelet-derived growth factor (alpha-PDGF) receptor mRNA. Southern blot analysis of the gene revealed amplification of the rearranged alpha-PDGF receptor gene in the glioma. A cDNA coding for an aberrant transcript from the amplified receptor gene was obtained and characterized. Partial nucleotide sequence analysis of the cDNA revealed a deletion of 243 nucleotides coding for 81 amino acids in a portion of the immunoglobulin-like domains of the extracellular region of the receptor. cDNA polymerase chain reaction (PCR) of the total cellular RNA in the glioma indicated that more than 80% of the transcripts have a deletion of 243 nucleotides. Analysis of a PCR-amplified DNA fragment derived from the amplified alpha-PDGF receptor gene in the glioma revealed that an exon coding for the 81 amino acids was removed by a 2.1 kb gene deletion. We also found amplification of the alpha-PDGF receptor gene in macroscopically normal cortex adjacent to the glioma from the same patient. The amplified gene in the macroscopically normal cortex has no major gene deletion, suggesting that gene amplification is not sufficient for the development of malignant gliomas.


Equivalent expression of paternally and maternally inherited WT1 alleles in normal fetal tissue and Wilms' tumours. Observations of non-random maternal 11p allele loss in Wilms' tumour (WT) have implied the possible involvement of an imprinted 11p locus in WT aetiology. A proposed 11p13 Wilms' tumour gene, WT1, has recently been isolated and encodes a zinc finger DNA-binding protein, the 3' untranslated region of which contains a polymorphic dinucleotide repeat (CA repeat) motif. We have exploited this transcribed CA repeat to examine the allelic expression pattern of WT1 and thereby determine whether transcriptional imprinting of this gene occurs. DNA and reverse-transcribed RNA from tumours and normal tissue were subjected to the polymerase chain reaction (PCR) using radiolabelled primers flanking the CA repeat. The gene was seen to be expressed from both of the constitutive alleles in 9-week human fetal kidney, all informative Wilm's tumours and neonatal kidney tissue adjacent to the tumours. In one tumour, known to be heterozygous for a point mutation in zinc finger 2, direct sequencing confirmed that both mutant and wild-type transcripts were being expressed. These results demonstrate that this gene is not subject to transcriptional imprinting in tumours or normal fetal kidney.


Skeletal muscle arises as a late event during development of wound sarcomas in v-jun transgenic mice. Mice carrying an H-2K-v-jun transgene develop malignant sarcomas by a multistage mechanism following wounding. Here we show that these malignancies are often heterogeneous in composition, containing both undifferentiated mesenchymal cells as well as focal areas of skeletal muscle. Such myogenic areas are not detectable in premalignant precursor lesions, suggesting that cells competent for muscle differentiation arise at a late stage of tumorigenesis. Immunohistochemical staining of transgenic sarcomas reveals that levels of v-Jun correlate inversely with muscle-specific gene expression, suggesting that high levels may be inhibitory to myogenesis. Consistent with this idea, we demonstrate that whereas high levels of v-Jun are able to block MyoD-dependent gene expression in vitro, the levels of v-Jun in sarcoma-derived myogenic cells are below the threshold required to produce this effect. The cell of origin of v-jun wound sarcomas, as well as the relationship between myogenic determination and multistage tumorigenesis, are discussed in the light of these results.


Sustained expression of the pim-1 kinase is specifically induced in myeloid cells by cytokines whose receptors are structurally related. We have examined the effects of myeloid growth factors on expression of the pim-1 kinase protein in human and murine myeloid cells. pim-1 protein was identified in K562 cells by immunoblotting as a 33 kDa protein. In the human factor-dependent myeloid leukemia cell line M07E, pim-1 protein was induced by interleukin 3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF), with maximum expression by 4 h. Expression continued for the duration of growth factor exposure, but declined rapidly when cytokines were removed. GM-CSF induced pim-1 protein in a dose-dependent manner, with expression being proportional to the proliferative effect of the cytokine. To examine the specificity of pim-1 protein induction, we compared pim-1 protein levels in myeloid cells which demonstrated different GM-CSF response phenotypes. We also examined the effects on pim-1 protein expression of different growth factors which induced similar response phenotypes. GM-CSF induced pim-1 protein in several myeloid cell lines, most of which demonstrated a proliferative response, but did not induce pim-1 protein expression in neutrophils or monocytic cells. In contrast, the murine cell line Mac-11 expressed pim-1 message in response to IL-3 and GM-CSF, but not in response to bryostatin or M-CSF, which were equivalent mitogens. In human U937 myeloid cells sustained expression of pim-1 protein was induced by GM-CSF, G-CSF and IL-6, but not by bryostatin. Expression of the pim-1 kinase protein in response to myeloid cytokines depends on both the nature of the growth factor and the response phenotype. The pim-1 kinase may be an important intermediate in transmembrane signaling or response phenotype induced by IL-3, GM-CSF and other cytokines whose receptors are structurally similar. Its constitutive expression in some myeloid leukemia cell lines suggests activation of signal cascades utilized by myeloid growth factors.


Inactivation of the remaining allele of the WT1 gene in a Wilms' tumour from a WAGR patient. A candidate gene (WT1) has recently been described for the 11p13 tumour-suppressor gene involved in the development of Wilms' tumour. This gene encodes a zinc finger protein which can bind to a specific DNA sequence. We have found a 226 base deletion in the mRNA from a unilateral Wilms' tumour, which would cause a frameshift that completely deletes the zinc finger domain. The tumour developed in a patient suffering from the WAGR syndrome, who had a constitutional 11p13 deletion, and so the 226 base deletion represents the inactivation of the remaining WT1 allele in the tumour. This provides further direct evidence that loss of function of WT1 is an essential step in the development of Wilms' tumour.


Chromosomal localization of human p85 alpha, a subunit of phosphatidylinositol 3-kinase, and its homologue p85 beta. The human phosphatidylinositol (PI) 3-kinase p85 alpha subunit gene and its homologue p85 beta were assigned to human chromosomes by analysis of their segregation in a panel of somatic cell hybrids using human-specific polymerase chain reaction primers. The p85 alpha locus was only present in hybrids retaining the human chromosome 5q. The presence of the p85 beta locus coincided with the presence of chromosome 19. The precise chromosomal sublocalization of these two genes was then determined by in situ hybridization. We confirmed the localization of the p85 alpha gene at 5q12-q13, as recently described (Cannizzaro, L.A., Skolnik, E.Y., Margolis, B., Croce, C.M., Schlesinger, J. & Huebner, K. (1991). Cancer Res., 51, 3818-3820) and positioned the p85 beta locus at 19q13.2-q13.4.


Surgical management of adenoid cystic carcinoma in the parotid gland. Although adenoid cystic carcinoma may be found in multiple sites in the head and neck as well as other glandular sites throughout the body, nowhere is management of the disease more controversial than in the parotid gland. Here the facial nerve is at risk from both the disease and the treatment. Seventy-five cases of adenoid cystic carcinoma of the parotid were analyzed. Patients were placed in four groups, depending on the type of parotid surgery received as definitive therapy: (1) lateral lobectomy, (2) total parotidectomy, (3) radical parotidectomy without preoperative facial weakness, and (4) radical parotidectomy with preoperative facial weakness. Patients were assessed with regard to staging of the initial lesion, the status of surgical margins, and the use of postoperative radiotherapy. The incidence of local recurrence and distant metastases were also recorded. Survival statistics are presented for each group. Though associated with facial nerve sacrifice, radical parotidectomy appears to offer clear advantages in terms of long-term disease-free survival in patients with T2 and T3 lesions. The residual facial paralysis may be rehabilitated primarily or secondarily to reduce patient morbidity. Four of 16 patients (25%) with preoperative weakness achieved 10-year survival when radical parotidectomy was used. Obtaining clear margins at the initial setting appears to offer improved survival.


Imaging rounds #111. VATER association.

Imaging rounds #111. VATER association. The following case describes the roentgenographic and clinical findings of a condition of interest to the orthopaedic surgeon. Initial history, physical findings, and roentgenographic examinations are found on this page. The next page presents the final clinical differential diagnosis.


[Use of silicon-impregnated compresses for echographic detection of section planes during hepatectomies]. It is essential to detect section planes during hepatectomy, notably for limited resections in cirrhotic patients. The authors describe a technical artifice, the use silicon-impregnated compresses, to help in the peroperative ultrasonographic detection of these section planes.


Reaction of cyanide with cytochrome ba3 from Thermus thermophilus: spectroscopic characterization of the Fe(II)a3-CN.Cu(II)B-CN complex suggests four 14N atoms are coordinated to CuB. Cytochrome ba3 from Thermus thermophilus reacts slowly with excess HCN at pH 7.4 to create a form of the enzyme in which CuA, cytochrome b, and CuB remain oxidized, while cytochrome a3 is reduced by one electron, presumably with the formation of cyanogen. We have examined this form of the enzyme by UV-visible, resonance Raman, EPR, and electron nuclear double resonance spectroscopies in conjunction with permutations of 13C- and 15N-labeled cyanide. The results support a model in which one CN- binds through the carbon atom to ferrous a3, supporting a low-spin (S = 0) configuration on the Fe; bridging by this cyanide to the CuB is weak or absent. Four 14N atoms, presumably donated by histidine residues of the protein, provide a strong equatorial ligand field about CuB; a second CN- is coordinated through the carbon atom to CuB in an axial position.


V beta 17 T-cell deletion by endogenous mammary tumor virus in wild-type-derived mouse strain. The wild-type-derived mouse strain PWK possesses a beta-chain variable region V beta 17a2 allele, which is expressed on mature T cells as part of the T-cell receptor of most mice expressing I-E, whereas V beta 17 T cells are deleted in all I-E+ laboratory mice bearing a V beta 17a1 allele. However, (PWK x CBA/J)F1 progeny and the wild-type-derived mouse strain MAI, which possesses the V beta 17a2 allele, display deletion of V beta 17 T cells. Analysis of (PWK x CBA/J) x PWK and of (PWK x MAI) x PWK backcrosses demonstrates that endogenous mouse mammary tumor virus MTV-6 from CBA/J and a MTV from strain MAI control the clonal deletion of V beta 17a2 as well as V beta 3 T cells. Furthermore, among I-E- progeny of a (MAI x C57BL/6) x C57BL/6 backcross, we observed that mice inheriting MTV of MAI have a reduced level of V beta 17 T cells, suggesting that the clonal deletion of V beta 17a2 T cells can be mediated in the absence of the I-E molecule. The 3' long terminal repeat of MTV MAI was cloned and translation of the open reading frame was compared to those of MTV known to encode superantigens. Comparisons indicate that MTV MAI has significantly diverged from the other MTVs. However, MTV MAI and MTV-6 share a stretch of 11 identical amino acids at the C terminus, which is divergent in MTV reacting with other V beta s. This suggests that this region is involved in determining the specificity toward V beta s and has been selectively conserved through evolution of the Mus species.


Initiator methionine tRNA is essential for Ty1 transposition in yeast. The yeast retrotransposon Ty1 transposes through an RNA intermediate by a mechanism similar to that of retroviral reverse transcription and integration. Ty1 RNA contains a putative minus strand primer binding site (-PBS) that is complementary to the 3' acceptor stem of the initiator methionine tRNA (tRNA(iMet)). Here we demonstrate that the tRNA(iMet) is used as a primer for Ty1 reverse transcription. Mutations in the Ty1 element that alter 5 of 10 nucleotides that are complementary to the tRNA(iMet) abolish Ty1 transposition, even though they are silent with regard to Ty1 protein coding. We have constructed a yeast strain lacking wild-type tRNA(iMet) that is dependent on a mutant derivative of tRNA(iMet) that has an altered acceptor stem sequence, engineered to restore homology with the Ty1 -PBS mutant. The compensatory mutations made in the tRNA(iMet) alleviate the transposition defect of the Ty1 -PBS mutant. The mutant and wild-type tRNA(iMet) are enriched within Ty1 virus-like particles irrespective of complementarity to the Ty1 -PBS. Thus, complementarity between the Ty1 -PBS and tRNA(iMet) is essential for transposition but is not necessary for packaging of the tRNA inside virus-like particles.


Rat brain expresses an alternatively spliced form of the dihydropyridine-sensitive L-type calcium channel alpha 2 subunit. We have cloned and sequenced a cDNA (rB-alpha 2) encoding an alpha 2 subunit of the voltage-sensitive L-type calcium channel (dihydropyridine receptor) of rat brain. The cDNA (3823 base pairs) encodes a protein of 1091 amino acids with a Mr of 123,822. The deduced amino acid sequence of rB-alpha 2 cDNA is highly similar (95% amino acid identity) to that of rabbit skeletal muscle alpha 2 subunit. The rB-alpha 2 protein is distinct from the previously cloned skeletal muscle alpha 2-subunit protein [Ellis, S. B., Williams, M. E., Ways, N. R., Brenner, R., Sharp, A. H., Leung, A. T., Campbell, K. P., McKenna, E., Koch, W., Hui, A., Schwartz, A. & Harpold, M. M. (1988) Science 241, 1661-1664] because it contains an insertion of 7 amino acid residues and a deletion of a 19-amino acid segment between putative transmembrane domains 1 and 2. We show that the rB-alpha 2 and skeletal muscle alpha 2-subunit transcripts are the variants produced by alternative splicing of the primary transcript and that they are differentially expressed in brain and skeletal muscle, respectively.


The gamma 1(34.5) gene of herpes simplex virus 1 precludes neuroblastoma cells from triggering total shutoff of protein synthesis characteristic of programed cell death in neuronal cells. The gamma 1(34.5) gene of herpes simplex virus 1 was previously shown to play a role in viral virulence since deletion of the gene reduced by a factor of approximately 100,000 the capacity of the virus to replicate in the central nervous system and cause mortality in the mouse. Here we show that in the human neuroblastoma cell line SK-N-SH of neuronal origin gamma 1(34.5) null mutants expressed early proteins, viral DNA, and mRNA of late genes. However, the onset of viral DNA synthesis triggered complete cessation of incorporation of radioactive precursors into proteins. The mutant and wild-type viruses replicated and could not be differentiated in cell lines or cell strains of nonneuronal origin. The results indicate that in the absence of the gamma 1(34.5) gene the SK-N-SH neuroblastoma cells triggered a response similar to the programed cell death of neuronal cells induced by metabolic stress. The gamma 1(34.5) protein precludes this cell response possibly in order to enable the protein synthesis necessary for viral replication.


Biochemical analysis of UV mutagenesis in Escherichia coli by using a cell-free reaction coupled to a bioassay: identification of a DNA repair-dependent, replication-independent pathway. Incubation of UV-irradiated plasmid DNA with a protein extract prepared from Escherichia coli cells led to the production of mutations in the cro gene residing on the plasmid. The mutations were detected in a subsequent bioassay step, which involved transformation of an indicator strain with the plasmid DNA that was retrieved from the reaction mixture, followed by plating on lactose/MacConkey plates. UV mutations produced in this cell-free reaction required the recA and umuC gene products and were prevented by rifampicin, an inhibitor of RNA polymerase, which inhibited plasmid replication. Removal of pyrimidine photodimers from the plasmid by enzymatic photoreactivation after the in vitro stage, but prior to transformation, increased plasmid survival as expected. Surprisingly, it also caused a large increase in the frequency of UV mutations detected in the bioassay. This photoreactivation-stimulated in vitro UV mutagenesis was dependent on the excision repair genes uvrA, uvrB, and uvrC and occurred in the absence of DNA replication. This suggests that two distinct UV mutagenesis pathways occurred in vitro: a replication-dependent pathway (type I) and a repair-dependent pathway (type II). DNA sequence analysis of type II UV mutations revealed a spectrum similar to that of in vivo UV mutagenesis. When the photoreactivation step was included in the protocol, type II UV mutagenesis did not require the RecA and UmuC proteins. These results are in agreement with the in vivo delayed photoreactivation phenomenon, where the removal of photodimers after an incubation period eliminated the requirement for RecA and UmuC in UV mutagenesis. The above system will enable the biochemical analysis of UV mutagenesis and the isolation of proteins involved in the process.


Transgenic indicator mice for studying activated retinoic acid receptors during development. Retinoic acid (RA) receptors (RARs) are ligand-inducible transcription factors that bind to specific DNA sequences associated with the regulatory regions of RA-regulatable genes. Since RA has been implicated as an important factor both in normal development and in teratological studies, one would like to have a model system that detects the presence of activated receptors during development. We have constructed a recombinant reporter gene that has three copies of the RA response element (RARE) from the RAR beta-2 promoter 5' to the herpes simplex virus thymidine kinase promoter; this regulatory region is coupled to the bacterial beta-galactosidase reporter gene. This construct was RA inducible in transient transfection assays in F9 embryonal carcinoma cells. Transgenic embryos with this reporter gene construct exhibited restricted and reproducible patterns of beta-galactosidase activity during embryogenesis, beginning between gestational ages day 7.5 and 8.5. At day 8.5, beta-galactosidase activity was detected in the closed neurotube and somites. Day 8.5 embryos, from pregnant females fed RA 14 hr earlier, exhibited a greater intensity and distribution of beta-galactosidase activity. Similarly, at later stages of gestation, maternal RA exposure resulted in enhanced embryonic beta-galactosidase expression. This type of transgenic indicator mouse should be useful in detailing the role of activated RARs during embryonic development.


Hypoxia injures endothelial cells by increasing endogenous xanthine oxidase activity. Exposure to decreasing oxygen tensions progressively increased xanthine dehydrogenase (XD) and xanthine oxidase (XO) activities over 48 hr in cultured pulmonary artery endothelial cells (EC) without altering XD/XO ratios. Increases in XD and XO activity in EC induced by hypoxia were associated upon reoxygenation with increased (P less than 0.05) extracellular superoxide anion (O2-.) levels that were inhibited by treatment with XO inhibitors (tungsten, allopurinol) or an anion-channel blocker (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid). EC monolayers subjected to hypoxia/reoxygenation also leaked more preloaded 51Cr, were more adherent to neutrophils, and permitted greater albumin transit than control monolayers. Treatment with tungsten, allopurinol, and/or superoxide dismutase decreased (P less than 0.05) 51Cr release, neutrophil adherence, and albumin transit in EC monolayers exposed to hypoxia/reoxygenation. We conclude that prolonged hypoxia increases both XO and XD activity in EC and may predispose the endothelium to oxidative and inflammatory damage.


Construction of small-insert genomic DNA libraries highly enriched for microsatellite repeat sequences. We describe an efficient method for the construction of small-insert genomic libraries enriched for highly polymorphic, simple sequence repeats. With this approach, libraries in which 40-50% of the members contain (CA)n repeats are produced, representing an approximately 50-fold enrichment over conventional small-insert genomic DNA libraries. Briefly, a genomic library with an average insert size of less than 500 base pairs was constructed in a phagemid vector. Amplification of this library in a dut ung strain of Escherichia coli allowed the recovery of the library as closed circular single-stranded DNA with uracil frequently incorporated in place of thymine. This DNA was used as a template for second-strand DNA synthesis, primed with (CA)n or (TG)n oligonucleotides, at elevated temperatures by a thermostable DNA polymerase. Transformation of this mixture into wild-type E. coli strains resulted in the recovery of primer-extended products as a consequence of the strong genetic selection against single-stranded uracil-containing DNA molecules. In this manner, a library highly enriched for the targeted microsatellite-containing clones was recovered. This approach is widely applicable and can be used to generate marker-selected libraries bearing any simple sequence repeat from cDNAs, whole genomes, single chromosomes, or more restricted chromosomal regions of interest.


Evidence for immune selection of hepatitis C virus (HCV) putative envelope glycoprotein variants: potential role in chronic HCV infections. E2/nonstructural protein 1, the putative envelope glycoprotein (gp72) of HCV, possesses an N-terminal hypervariable (E2 HV) domain from amino acids 384 to 414 of unknown significance. The high degree of amino acid sequence variation in the E2 HV domain appears to be comparable to that observed in the human immunodeficiency virus type 1 gp120 V3 domain. This observation and the observation that the HCV E2 HV domain lacks conserved secondary structure imply that, like the V3 loop of human immunodeficiency virus 1 gp120, the N-terminal E2 region may encode protective epitopes that are subject to immune selection. Antibody-epitope binding studies revealed five isolate-specific linear epitopes located in the E2 HV region. These results suggest that the E2 HV domain is a target for the human immune response and that, in addition to the three major groups of HCV, defined by nucleotide and amino acid sequence identity among HCV isolates, E2 HV-specific subgroups also exist. Analysis of the partial or complete E2 sequences of two individuals indicated that E2 HV variants can either coexist simultaneously in a single individual or that a particular variant may predominate during different episodes of disease. In the latter situation, we found one individual who developed antibodies to a subregion of the E2 HV domain (amino acids 396-407) specific to a variant that was predominant during one major episode of hepatitis but who lacked detectable antibodies to the corresponding region of a second variant that was predominant during a later episode of disease. The data suggest that the variability in the E2 HV domain may result from immune selection. The findings of this report could impact vaccine strategies and drug therapy programs designed to control and eliminate HCV.


GLUT4 facilitates insulin stimulation and cAMP-mediated inhibition of glucose transport. The glucose transporter isoform GLUT4 is found only in cells that exhibit insulin-sensitive glucose transport. To investigate the function of this transporter, L6 myoblasts were stably transfected with GLUT4 cDNA. GLUT4 underwent insulin-dependent movement to the cell surface in myoblasts overexpressing the transporter. One cell line (243-6) expressed sufficient levels of the GLUT4 protein to study insulin-dependent glucose transport. Unlike wild-type L6 cells, 243-6 myoblasts exhibited two features that are characteristic of differentiated muscle fibers and adipocytes in vivo: a large insulin-stimulated component of glucose transport and inhibition of this stimulated component by cAMP. Relative to normal L6 cells, 243-6 cells responded to insulin or insulin-like growth factor 1 with a 5-fold larger increase in 2-deoxy[3H]glucose uptake. N6,O2'-Dibutyryladenosine 3',5'-cyclic monophosphate (Bt2cAMP) did not inhibit transport in normal L6 myoblasts, which express only GLUT1, but inhibited IGF-1/insulin-stimulated transport by 50% in 243-6 cells. The effect of cAMP was investigated further by using Chinese hamster ovary cells transiently expressing GLUT1 and GLUT4. Bt2cAMP inhibited glucose transport only in Chinese hamster ovary cells expressing GLUT4. These results indicate that cAMP-mediated inhibition of glucose transport is dependent on expression of the GLUT4 isozyme.


On the expression and regulation of 5-lipoxygenase in human lymphocytes. The expression of arachidonate 5-lipoxygenase (arachidonate:oxygen 5-oxidoreductase, EC 1.13.11.34) and the 5-lipoxygenase-activating protein (FLAP) genes in human tonsillar B cells and lymphoblastoid B-cell lines was demonstrated at the transcriptional level by reverse transcription-PCR analysis. Also, five lymphoblastoid T-cell lines were investigated and found to express the FLAP gene but not the 5-lipoxygenase gene, suggesting that the transcriptional regulation of these two genes is different. Western blot analysis of the cytosolic proteins from a lymphoblastoid B-cell line with an antiserum raised against purified human leukocyte 5-lipoxygenase revealed an immunoreactive band that comigrated with recombinant human 5-lipoxygenase. Intact B cells produced very low amounts of leukotriene B4 and 5-hydroxyeicosatetraenoic acid upon stimulation with the calcium ionophore A23187 and arachidonic acid, in comparison to the amounts formed by sonicates of these cells. However, preincubation of intact lymphoblastoid B cells with the glutathione-depleting agents azodicarboxylic acid bis(dimethylamide) or 1-chloro-2,4-dinitrobenzene prior to the addition of the calcium ionophore A23187 and arachidonic acid led to similar amounts of leukotriene B4 as were formed by sonicated cells. In contrast, the glutathione synthesis inhibitor buthionine sulfoximine diminished the cellular level of glutathione by greater than 90% but did not influence the production of leukotriene B4 or 5-hydroxyeicosatetraenoic acid in intact cells. These results demonstrate that certain drugs affecting the redox status can stimulate the cryptic 5-lipoxygenase activity in intact lymphoblastoid B cells but that the mechanism of this activation is unclear and appears not to be directly related to intracellular glutathione levels.


Parallel pathways in macaque monkey striate cortex: anatomically defined columns in layer III. Visual information reaching striate cortex comes from parallel pathways, and the information is organized, or processed, by the layers and columns of striate cortex. To better understand how this is accomplished anatomically, we asked whether parallel pathways originating in the lateral geniculate nucleus (LGN), and terminating separately in layer IV, remain separate in layer III of macaque monkeys. Layer III is of interest since it may play a special role in color and form vision but not in analysis of visual motion. The chief finding was that cells in "blobs" of layer III that stain densely for cytochrome oxidase receive indirect input, via layer IVC, from both LGN magnocellular (M) and parvocellular (P) cells. This is important because the P and M pathways may represent color/form and motion-processing channels, respectively. Interblob cells receive indirect input, via layers IVC and IVA, from the LGN P cells. Also, as suggested by others, our data demonstrate that layer III can be subdivided. The bottom tier, layer IIIB, receives direct projections from all cortical layers. Output from layer IIIB appears to remain intrinsic to striate cortex. In contrast, the top tier, layer IIIA, receives projections from layer IIIB as well as from layers IVA, IVB (blobs only), and V, but it receives no direct projections from LGN recipient layers IVC and VI. Unlike layer IIIB, the output of layer IIIA reaches extrastriate areas. Thus, impulses arriving from parallel LGN pathways may be recombined through serial stages in striate cortex to produce a set of parallel pathways that are qualitatively different from the original LGN set.


In vivo labeling of L-type Ca2+ channels by fluorescent dihydropyridines: evidence for a functional, extracellular heparin-binding site. We have synthesized and characterized fluorescently labeled dihydropyridines (DHPs) as probes for L-type Ca2+ channels. Racemic as well as (+)- and (-)-1,4-dihydro- 2,6-dimethyl-4-(2-trifluoromethylphenyl)-3,5-pyridinecarboxylic acid 2-(aminoethyl)ethyl ester hydrochlorides were coupled to boron dipyrromethane (Bodipy) derivatives. (4,4-Difluoro-5,7-dimethyl-4-bora-3a,4a-diaza)-3- (s-indacene)propionic acid (DMBodipy)-DHP and (4,4-difluoro-7-styryl-4-bora-3a,4a-diaza)-3-(s-indacene+ ++)propionic acid (STBodipy)-DHP have Kd values in the nanomolar range for membrane-bound or partially purified skeletal muscle and for neuronal L-type Ca2+ channels. (-)- and (+)-STBodipy-DHPs block 45Ca2+ uptake through L-type Ca2+ channels into GH3 cells with IC50 values of 14.8 and 562 nM, respectively. The measurement of bound fluorescence after removal of free DMBodipy-DHP with charcoal shows that the probes can substitute for radioactive ligands to study the properties (equilibrium binding, kinetics, allosteric regulation) of partially purified L-type Ca2+ channels from skeletal muscle. L-type Ca2+ channels on GH3 cells were steroselectively visualized by using the optical enantiomers of STBodipy-DHP. Heparin inhibited GH3 cell labeling by (-)-STBodipy-DHP with an IC50 value of 9.7 micrograms/ml and blocked L-type Ca(2+)-channel-mediated 45Ca2+ uptake with an IC50 value of 32 micrograms/ml. These findings argue for an extracellular orientation of the heparin-binding domain of the Ca2+ channel that is coupled to the DHP receptor.


DNA binding and transcriptional regulatory activity of mammalian achaete-scute homologous (MASH) proteins revealed by interaction with a muscle-specific enhancer. The MASH genes are vertebrate homologues of achaete-scute, genes required for neuronal determination in Drosophila. The sequence of MASH1 and MASH2 contains a basic helix-loop-helix (bHLH) motif that is present in other transcriptional regulators such as MyoD and E12. In the absence of an authentic target for the MASH proteins, we examined their DNA binding and transcriptional regulatory activity by using a binding site (the E box) from the muscle creatine kinase (MCK) gene, a target of MyoD. Like myogenic bHLH proteins, the MASH proteins form heterooligomers with E12 that bind the MCK E box with high affinity in vitro. Unexpectedly, however, MASH1 and MASH2 also activate transcription of both exogenous and endogenous MCK in transfected C3H/10T1/2 fibroblasts. However, they do not induce myogenesis. Myogenic activity is not exclusively a property of the MyoD basic region, as substitution of this domain fails to confer myogenic activity on MASH1. These data suggest that different bHLH proteins may activate overlapping but distinct sets of target genes in the same cell type.


The synaptic vesicle protein synaptotagmin associates with calcium channels and is a putative Lambert-Eaton myasthenic syndrome antigen. Immunoglobulin G fractions from patients with Lambert-Eaton myasthenic syndrome (LEMS), an autoimmune disease of neuromuscular transmission, immunoprecipitate 125I-labeled omega-conotoxin GVIA-labeled calcium channels solubilized from rat brain. A 58-kDa antigen was detected by probing Western blots of partially purified calcium channels with LEMS plasma and IgG and was shown to be the relevant antigen in omega-conotoxin receptor immunoprecipitation. Monoclonal antibody 1D12, produced by immunizing mice with synaptic membranes, has properties similar to these autoimmune IgGs in both immunoprecipitation and Western blotting assays. 1D12 antigen was purified by immunoaffinity chromatography and shown to bind LEMS IgG. The antigen was identified by screening a rat brain cDNA library with 1D12 and was found to have strong homology to the synaptic vesicle membrane protein synaptotagmin. Our results indicate therefore that these antibodies immunoprecipitate omega-conotoxin receptors by binding to synaptotagmin that is associated with calcium channels. We suggest that the interaction between synaptotagmin and the voltage-gated calcium channel plays a role in docking synaptic vesicles at the plasma membrane prior to rapid neurotransmitter release and that autoantibody binding to a synaptotagmin-calcium-channel complex may be involved in the etiology of LEMS.


Human nucleotide excision nuclease removes thymine dimers from DNA by incising the 22nd phosphodiester bond 5' and the 6th phosphodiester bond 3' to the photodimer. By using a human cell-free system capable of nucleotide excision repair, a synthetic substrate consisting of a plasmid containing four thymidine dimers at unique locations, and deoxyribonucleoside 5'-[alpha-thio]triphosphates for repair synthesis, we obtained DNA fragments containing repair patches with phosphorothioate linkages. Based on the resistance of these linkages to digestion by exonuclease III and their sensitivity to cleavage by I2, we were able to delineate the borders of the repair patch to single-nucleotide resolution and found an asymmetric patch with sharp boundaries. That the repair patch was produced by filling in a gap generated by an excision nuclease and not by nick-translation was confirmed by the finding that the thymidine dimer was released in a 27- to 29-nucleotide oligomer.


[Chemoembolization of hepatocellular carcinoma in cases of isolated liver involvement]. Chemoembolization is an effective treatment for hepatocellular carcinoma, giving results equally as good as surgical therapy for T2 tumours. Survival can be prolonged and side-effects can be reduced by combining Lipiodol and Gelfoam for chemoembolization, employing a modified technique, with repeated procedures, and using appropriate follow-up treatment. The toxicity of the procedure is acceptable, but it requires supportive therapy necessitating an intense interdisciplinary co-operation.


Evaluation of hypertension screening in the Hutterite population. The purpose of this study was to examine the impact of a hypertension screening program on Hutterites from Alberta, Canada who were found to have a diastolic blood pressure greater than 90 mm Hg. At approximately 16 months postscreening, 200 subjects completed a self-administered questionnaire requesting information about physician follow-up and pharmacologic and nonpharmacologic therapy. Seventy-four percent of the subjects reported that they had their blood pressure measured at least twice since screening. A third of the respondents reported changes in pharmacologic management. Subjects reported weight loss (36%), and salt and alcohol reduction (64% and 48%, respectively) as nonpharmacologic methods for lowering blood pressure. In terms of physician follow-up and pharmacologic and nonpharmacologic modification, the screening program was effective.


[Adrenergic control of myocardial contractile function in rats with incorporated 137Cs]. It was shown on isolated myocardium preparations that internal gamma irradiation of rats with 137Cs (1.6 MBq/kg) decreased the contractile function of ventricles and had no essential effect on the auricle contractility. The inotropic response of myocardium to activation of alpha-adrenergic receptors decreased, and of beta-adrenergic receptors decreased.


The effects of noise stress on leukocyte function in rats. It has been reported that exposure to increased noise levels impairs wound healing in surgical patients and in rats. The purpose of the present study was to determine if exposure to noise stress would alter the biological function of neutrophils, macrophages, and lymphocytes, leukocytes that are involved in wound healing. Rats were exposed to 80 db of "rock" music for 24 hr, during which time the control animals were maintained in their usual environment. Leukocyte subpopulations were obtained and stimulated in vitro. Neutrophils and macrophages from noise-exposed animals secreted significantly less superoxide anion and interleukin-1 than cells from control animals. Lymphocyte function was not altered following noise stress. We conclude that short-term exposure of rats to noise stress alters some of the biological functions of leukocytes.


[The spectral and paramagnetic properties of oxyhemoglobin solutions UV-irradiated in the presence of ascorbic acid]. Absorption spectra and ESR of aqueous and aqueous/glyceric solutions of oxyhemoglobin exposed to UV radiation (250-400 nm) at 293 and 77 K in the presence of ascorbic acid have been analyzed. Vitamin C (5 x 10(-5) M) has been shown to exert a photoprotective effect with regard to oxyhemoglobin (2 x 10(-6) M) UV-irradiated with a dose of 0.86 x 10(5) J/m2 at 293 K. The photoprotective effect of ascorbic acid is also displayed after UV irradiation of frozen (77 K) aqueous/glyceric oxyhemoglobin solutions (2.53 x 10(-5) M). It is concluded that ascorbic acid can be a scavenger with respect to active UV-induced particles in protein systems, including O2-. and OH. Proposed is a mode of processes leading to UV inactivation of hemoprotein molecules.


[The characteristics of the realization of cytogenetic damage in mammalian and plant cells exposed to low doses of radiation]. A complicated character of the cytogenetic injury dependence upon radiation dose was revealed after low-level gamma irradiation of Vicia faba seedlings and Chinese hamster fibroblasts. The dependence was linear with low-level secondary exposure to 70 GeV protons. The authors discuss a threshold nature of induction of the cytogenetic damage repair responsible for a high outcome of damages under the effect of low-level gamma radiation.


[A phase II trial of pirarubicin in untreated disseminated small cell lung cancer. A cooperative study of the French Pneumo-Cancerology Group]. The usual form of chemotherapy of metastatic small cell lung cancer gives a 50% objective response with a mean survival of 7-8 months. We have tested a new antimitotic drug using pirarubicin alone in 26 patients. After the second treatment we noticed a response level of 12% with moderate toxicity. Then, we undertook classical chemotherapy using cisplatin-V16. After 3 doses the response level was 50% with a median survival of 32 weeks. In our study the use of a single drug pirarubicin in metastatic small cell cancer did not appear to worsen the chance of survival in patients if polychemotherapy was carried out immediately in cases which failed on the single drug. Our monotherapy did not appear to induce resistance to affective polychemotherapy. This method applied carefully to patients with metastatic disease with a strict follow up may be utilised in the assessment of the efficacy of the newer antimitotic drugs.


In vitro glycated low-density lipoprotein interaction with human monocyte-derived macrophages. Human low-density lipoprotein (LDL) was glycated in vitro (5 days, glucose 50 mmol/l), labelled with 125I, and its binding and uptake by human monocyte-derived macrophages studied. Glycation produced lower binding and lower uptake. Competition experiments using unlabelled LDL (control, glycated, and acetyl-LDL) showed that most glycated LDL was taken up by the apolipoprotein-B100: E receptor pathway. Results suggest that less of the glycated LDL may enter the cells via scavenger receptors, and very minute amount via non-saturable receptor-independent pathways.


Lipopolysaccharide suppresses cytokine release from coxsackie virus-infected human monocytes. Infections by coxsackie virus B3 (CVB3) have been reported to be associated with an enhanced influx of mononuclear leukocytes into afflicted tissue. Current evidence indicates that monocytes/macrophages are specifically involved in CVB3-induced myocarditis by maintaining a chronic inflammatory response. To examine susceptibility and reactivity to CVB3, freshly isolated human monocytes were exposed to various virus doses (0.1-10 MOI) in the presence or absence of macrophage-activating lipopolysaccharide (LPS). CVB3 infection alone induced an activation of monocytes as evidenced by enhanced adherence, release of cytokines and secretion of prostaglandin E2 (PGE2). Simultaneous addition of LPS almost entirely suppressed LPS-specific production of tumour necrosis factor alpha (TNF alpha) and PGE2, partially inhibited release of interleukin 1 beta (IL 1 beta) and did not affect interleukin 6 (IL6) synthesis of CVB3-infected monocytes. These data show that CVB3 activates monocytes to cytokine production but renders them unreactive to further activating stimuli. Further studies should determine the extent to which continuous cytokine release from persistently CVB3-infected monocytes, and their apparent unresponsiveness to other stimuli, contribute to chronic myocarditis.


A histopathologic study of the pigmented fundus lesions in familial adenomatous polyposis. A postmortem examination of the eyes of a 61-year-old woman with familial adenomatous polyposis was performed using light microscopy and transmission and scanning electron microscopy. Numerous lesions of the retinal pigment epithelium (RPE) were identified, which had one of three basic configurations: a monolayer of hypertrophic cells, a mound of RPE cells interposed between the RPE basement membrane and the inner collagenous layer of Bruch's membrane, or a multilayered mound of hyperplastic cells. The presence of abnormal pigment granules in cells within the lesions and cells from areas of grossly normal RPE indicates a generalized defect in melanogenesis. Although the pigmented fundus lesions of familial adenomatous polyposis are often referred to clinically as congenital hypertrophy of the RPE, the prominent component of cellular hyperplasia more appropriately designates them as hamartomas of the RPE. Their development is likely the result of the same loss of regulatory control of cell growth and replication that gives rise to the multiple colorectal polyps and soft tissue tumors that characterize this condition.


[Electrophysiological investigation of polyneuropathies]. Needle electromyography, nerve conduction velocity measurement and recording of T, H or F waves, are the most commonly used methods to investigate patients with polyneuropathies. The grade, distribution, and functional type of the lesions are to be determined. A classification is proposed according to acute or chronic character of the neuropathy course and to prevailing signs of demyelination or axonal degeneration.


[Diagnostic approach].

[Diagnostic approach]. Multiple mononeuropathies and peripheral polyneuropathies are due to a large variety of causes, and they frequently raise difficult diagnostic problems. Clinical examination has lost nothing of its value, but electrophysiological techniques and, in some cases, neuromuscular biopsy provide a better approach to the diagnosis of peripheral neuropathies. These neuropathies can now be classified according to their acute, subacute or chronic course and to the axonal or demyelinating nature of the physiopathological process involved.


[Multifocal neuropathy with persistent conduction block]. Multifocal neuropathy, recently individualized, is characterized by progressive, asymmetrical lesions predominant in the upper limbs and associated with multifocal and persistent conduction block. Two clinical presentations have been identified: sensorimotor multineuritis and pure motor form which may mimick an anterior grey horn disease. The pathophysiology of the syndrome and the significance of its frequent association with the antibody to GM1 ganglioside remain debated. Various treatments have been tried with inconstant results.


[Sensory neuropathies].

[Sensory neuropathies]. This review paper analyzes the different steps in the diagnosis of peripheral sensory neuropathies. Although electrodiagnostic tests are almost invariably needed, other investigations should be performed according to an accurate prior clinical evaluation. Some causes are frequent and easy to discover, such as diabetes and chronic renal failure. On the opposite, genetically determined and dysimmune neuropathies are less frequent, and may go unrecognized for a long time. A careful survey of the clinical, biological and electrophysiological features sometimes discloses a specific aetiology of an initially unclassified neuropathy.


[Polyneuropathies and monoclonal gammopathies, so-called benign]. As all other monoclonal dysglobulinaemias, the allegedly benign monoclonal dysglobulinaemia (also called of undetermined nature) can be responsible for peripheral neuropathy. It must be noted that electrophoresis and immuno-electrophoresis may be insufficient to demonstrate its presence and that this can be done with specific immunological techniques, such as ELISA or immunoblot). Once the monoclonal gammopathy has been recognized, its relation to the peripheral nerve lesion is not always easy to prove. Antibody activity against certain constituents of the nerve can sometimes be evidenced, notably for IgM. It is interesting to point out that almost all IgM-related neuropathies have highly characteristic clinical, histological and immunological features. Various other mechanisms, often still unknown, are most probably involved.


[Therapeutic approach in peripheral neuropathies]. On the whole, the treatment of peripheral neuropathies is disappointing, even when a cause has been identified, because nerve lesions, and particularly axonal diseases, are frequently severe, and the mechanisms or peripheral nerve fibre repair is very slow. However, during the last few years important advances have been made in the treatment of acute and chronic acute inflammatory demyelinating polyneuropathies, most probably of dysimmune origin, with plasma exchanges and, more recently, intravenous human plasma immunoglobulins. Therapeutic trials are in progress in neuropathies associated with monoclonal gammopathies, notably IgM. In diabetic neuropathy numerous studies are going on, in particular with aldolase reductase inhibitors and with gangliosides.


[Peripheral neurologic manifestations of infection by the human immunodeficiency virus]. Peripheral nerve lesions observed in the progressive immunodeficiency associated with infection are remarkable for their diversity and their potential severity. At the onset, demyelinating neuropathies predominate, often with signs of atypical polyradiculoneuritis associated with inflammatory lesions or vasculitis sometimes of the necrotizing type, as that observed in periarteritis nodosa. Later on, at the immunosuppression stage, axonal lesions predominate. At the AIDS stage one may find lymphomatous infiltration of nerves and nerve roots and, chiefly, opportunistic cytomegalovirus (CMV) infection. This infection produces meningoradiculitis, mainly in the territory of the cauda equina, or multifocal neuropathy frequently associated with CMV retinitis. Early treatment of CMV neuropathies may stabilize the lesions or even result in functional improvement, but the overall prognosis of late neuropathies remains poor.


[Encephalopathies in infection by human immunodeficiency virus]. Encephalitis occupies a large part in the neurological complications of HIV infection. It is frequent and in most cases of poor prognosis. Some cases of encephalitis are directly related to HIV while others are caused by an opportunistic infection. Among the former is the acute encephalitis coincident with seroconversion, which is exceptional and spontaneously regressive, and the subacute encephalitis better known as HIV encephalopathy which has a constantly pernicious course ending in subcortical dementia lethal within a few months. Some cases of opportunistic encephalitis are associated with a virus: a Papovavirus is responsible for progressive multifocal leucoencephalopathy where mental deterioration is combined with focal symptoms, both resulting in death in less than 6 months. Cytomegalovirus is responsible for an encephalitis that is frequently found on pathological examination but is usually subclinical. Anecdotic cases of toxoplasmic encephalitis have been reported. Finally, emphasis should be placed on the frequency of encephalitis-associated pathologies with all possible combinations, the most common being HIV encephalitis with another encephalitis and/or focal ou multifocal infectious or tumoral processes.


[Diarrhea in AIDS. Group AIDS GIT].

[Diarrhea in AIDS. Group AIDS GIT]. Gastrointestinal (GI) infections are frequent in AIDS patients. The frequency and type of opportunistic GI infections are exactly the same in homosexuals and heterosexuals. Diarrhoea is the usual sign of GI infection, and its mechanism seems to combine a secretory component and a malabsorption. Although a number of pathogens can be isolated, in many cases the diarrhoea cannot be explained by an infection or a lesion. The hypothesis of a primary HIV infection in the epithelium of the small bowel and colon has not been confirmed by immunofluorescence and molecular hybridization. The HIV virus has been found in the GI mucosa, but it was probably carried by the immune cells in general circulation (CD4 lymphocytes and macrophages) which subsequently colonize the chorion of the mucosa.


Participation of non-zinc finger residues in DNA binding by two nuclear orphan receptors. Steroid-thyroid hormone receptors typically bind as dimers to DNA sequences that contain repeated elements termed half-sites. NGFI-B, an early response protein and orphan member of this receptor superfamily, binds to a DNA sequence that contains only one half-site (5'-AAAGGTCA-3'). A domain separate from the NGFI-B zinc fingers, termed the A box, was identified and is required for recognition of the two adenine-thymidine (A-T) base pairs at the 5' end of the NGFI-B DNA binding element. In addition, a domain downstream of the zinc fingers of the orphan receptor H-2 region II binding protein, termed the T box, determined binding to tandem repeats of the estrogen receptor half-site (5'-AGGTCA-3').


Body-wall muscle formation in Caenorhabditis elegans embryos that lack the MyoD homolog hlh-1. The myoD family of DNA binding proteins has been implicated in the control of myogenesis in a variety of organisms. Searches for homologs in the nematode Caenorhabditis elegans yielded only one gene, designated hlh-1, expressed in body-wall muscle cells and their precursors. To assess the role of hlh-1 in C. elegans myogenesis, genetic deficiencies spanning the hlh-1 locus were isolated after gamma irradiation. Embryos homozygous for these deficiencies exhibited extensive body-wall muscle differentiation, including expression of several characteristic myofilament proteins and weak contracile behavior. Thus, zygotic hlh-1 expression was not required for body-wall muscle precursors to adopt muscle cell fates.


Lithium-sensitive production of inositol phosphates during amphibian embryonic mesoderm induction. Mesoderm induction and body axis determination in frog (Xenopus) embryos are thought to involve growth factor-mediated cell-cell signaling, but the signal transduction pathways are unknown. Li+, which inhibits the polyphosphoinositide (PI) cycle  signal transduction pathway in many cells, also disrupts axis determination and mesoderm induction. Amounts of the PI cycle-derived second messenger, inositol 1,4,5-trisphosphate, increased during mesoderm induction in normal embryos; addition of Li+ inhibited the embryonic inositol monophosphatase and reversed this increase. Embryonic PI cycle activity thus shows characteristics that indicate it may function in mesoderm induction and axis determination.


Calcium-regulated phosphorylation within the leucine zipper of C/EBP beta. Alterations in intracellular calcium levels activate several signal transduction pathways resulting in distinct patterns of gene expression. Here, a pathway for calcium-mediated signals is demonstrated that involves C/EBP beta, a member of the bZip family of transcription factors. In pituitary cells C/EBP beta was phosphorylated in response to increased intracellular calcium concentrations as a consequence of the activation of a calcium-calmodulin-dependent protein kinase. Phosphorylation of serine at position 276 within the leucine zipper of C/EBP beta appeared to confer calcium-regulated transcriptional stimulation of a promoter that contained binding sites for C/EBP beta.


Case report 714. Postirradiation osteosarcoma after radiation of metastatic skeletal lesion. Two cases of postirradiation osteosarcoma are presented--one in a 76-year-old woman with breast carcinoma and subsequent osteosarcoma after radiation therapy for a metastatic lesion in the right tibia, and the other in a 16-year-old girl with hepatocellular carcinoma metastatic to the left tibia and osteosarcoma after radiation therapy to that bone. Microscopically, both cases were high-grade spindle cell lesions with osteoid production. Both patients fared poorly. This is a rare complication of radiation therapy.


Case report 716. Soft-tissue metastasis in synovial sarcoma. We describe the case of a 21-year-old woman with a synovial sarcoma of the thigh, who exhibited a soft-tissue metastasis to the shoulder approximately 3 months after her initial presentation. Due to its increasing availability and the recognized importance of MRI in the preoperative evaluation of patients with soft-tissue masses, it is important to be aware of even the uncommon manifestations of frequently encountered soft-tissue sarcomas.


Case report 720. Multicentric mixed tumor of bone with pulmonary involvement. A man presented with a mass in the left first metacarpal bone. Later, his chest radiograph showed extensive, bilateral, rounded opacities in both lungs with enlarged hilar lymph nodes, and he developed expanding lesions in the left radius, ulna, and metacarpal bones. The pulmonary lesions were treated with radiotherapy and cytotoxic agents, and the tumor mass in the first metacarpal was debulked. All biopsies showed similar features of a mixed tumor (pleomorphic adenoma) with metastatic or embolic pulmonary involvement; ultrastructural and immunocytochemical investigations supported this unique diagnosis. The patient remains well 15 years after the initial diagnosis. It is possible that the myoepithelial elements in this case had been displaced intraosseously during development. We are not aware of a similar case in the literature.


Central pontine myelinolysis as a manifestation of the paraneoplastic syndrome. We have described a patient who had lung carcinoma, syndrome of inappropriate antidiuretic hormone, and central pontine myelinolysis (CPM). Although this association is well-known, it appears to be the first report having radiographic documentation of both the intracranial and intrathoracic abnormalities. When searching for the cause of CPM, the possibility of an underlying malignancy should also be considered. This may be more common than the radiology literature would suggest.


Evaluation of cervical cord function using spinal evoked potentials from surface electrode. Spinal evoked potentials from cervical skin surface (surface spinal evoked potentials) were measured to evaluate spinal cord function as a convenient method that precludes inserting electrodes into the epidural space, and results were compared with those of the former epidural recording method. Surface spinal evoked potentials were obtained from cervical skin surface over the C3, C5, and C7 spinous processes after median nerve stimulation in 18 normal subjects and 37 patients with a cervical lesion. In normal subjects, surface spinal evoked potentials consisted of three negative waves (N1, N2, N3). Abnormal N2 (80%) and abnormal N3 (100%) were observed in cervical myelopathy, and abnormal N2 was observed only in radiculopathy; this allows for differentiation between myelopathy and radiculopathy. Comparing preoperative and postoperative surface spinal evoked potentials, it was seen that improvement of clinical symptoms was proportional to that of surface spinal evoked potentials.


Spinous process-splitting laminoplasty using hydroxyapatite spinous process spacer. Between February 1986 and August 1989, 45 patients with cervical myelopathy were treated by spinous process-splitting laminoplasty. Hydroxyapatite intraspinous spacers were used to maintain the enlargement of the cervical spinal canal. The shape of this spacer is trapezoidal. After sagittal splitting of the spinous process, the spacer was inserted between the two halves and affixed with the wire. Histologic study showed there was good fusion between the spacer and bone. In all cases, good enlargement of the cervical spinal canal was achieved. Spacer displacement, wire breakage, and postoperative infection were not seen. There was no postoperative neurologic deterioration. Computed tomography showed that the width of the cervical spinal canal was maintained.


Lipid peroxidation and loss of potassium from red blood cells produced by phototoxic quinolones. Alterations of the cationic permeability of red blood cell membranes induced by the photosensitiser nalidixic acid were demonstrated by evaluating the potassium loss from intact erythrocytes. The results show that an increase in intracellular potassium efflux, precedes the photohemolysis induced by nalidixic acid. The addition of a nonpermeable osmotic solute, such as sucrose, inhibited photohemolysis but not the potassium loss, indicating a colloid osmotic lysis. Lipid peroxidation induced by nalidixic acid and other photosensitiser quinolones (oxolinic acid and rosoxacin) was time irradiation-dependent. Although rosoxacin was the most photoperoxidative, none of the three quinolones studied produced significant lipid peroxidation. However, of the three quinolones studied, only rosoxacin considerably diminished the percentage of the cholesterol extracted from red blood cell membranes. It is postulated that the increased cation permeability induced by nalidixic and oxolinic acids cannot be attributed to cholesterol oxidation nor to lipid peroxidation; a more probable mechanism is photo-oxidation of amino acid residues of the membrane proteins. However, the lysis induced by rosoxacin is caused by photo-oxidation of cholesterol, not excluding other cellular targets.