Chronic low-dose alprazolam augments gamma-aminobutyric acid(A) receptor function. After acute administration of low doses, alprazolam displays unusual behavioral and neurochemical characteristics. To determine whether chronic low-dose alprazolam has unique effects, we treated mice for 1-14 days with alprazolam 0.2 mg/kg per day and evaluated open-field activity, benzodiazepine receptor binding, t-butylbicyclophosphorothionate binding, and muscimol-stimulated chloride uptake. Open-field activity in treated mice was similar to that of control mice at each timepoint during alprazolam administration. Similarly, benzodiazepine receptor binding in vivo was unchanged in five brain regions. Benzodiazepine receptor binding in vivo was unchanged in five brain regions. Benzodiazepine binding in vitro in the cortex was unaffected by alprazolam treatment, as was t-butylbicyclophosphorothionate binding in the cortex. However, muscimol-stimulated chloride uptake was increased after 2 and 4 days of alprazolam compared with results after 1, 7, and 14 days. These results are consistent with prior reports of unusual effects of low-dose alprazolam and extend these findings to chronic administration.


Detoxification for triazolam physical dependence. The clinical characteristics of a series of patients who developed physical dependence to triazolam has not previously been described. We report five cases of high-dose triazolam abuse in which the daily dosage ranged from 5 to 15 mg (100-300 mg diazepam equivalent) and the duration of use ranged from 3 months to 5 years. Physical dependence usually occurs in the context of a history of alcohol or other drug abuse, a history of panic attacks or anxiety disorder, substitution of triazolam for a longer-acting benzodiazepine, or inappropriate use of triazolam as an anxiolytic agent given as multiple daily doses. A rational approach to the pharmacotherapy of triazolam withdrawal is described. Treatment to replace the triazolam with a long half-life CNS depressant and strategies for this pharmacologic therapy are discussed. The abuse liability of triazolam may be no greater and is most likely less than that of some other benzodiazepines.


Competition between an aphakic and an occluded eye for territory in striate cortex of developing rhesus monkeys: cytochrome oxidase histochemistry in layer 4C. Monkey models were used to examine the effects of competition for cortical territory between two eyes which were deprived simultaneously, but each eye experienced a different type of deprivation. We wanted to determine whether, under this condition of binocular unequal deprivation, the postnatal process of segregation into ocular dominance columns proceeds according to the same rules as those that apply to competition between a deprived and an undeprived fellow eye. Our models involved surgical removal of the natural lens from one eye in newborn rhesus monkeys. The resulting aphakia was corrected optically to a near point with extended-wear contact lenses. The fellow eyes were either left unmanipulated or occluded with opaque contact lenses for varying periods during the day. At the end of the rearing period, some monkeys from each experimental group had either one eye enucleated or sustained injury to the retinal ganglion cells of one eye. The histochemical reaction for cytochrome oxidase was used to reveal the widths of ocular dominance columns in layer 4C of striate cortex in these monkeys. Under all experimental conditions, the axons related to the two eyes occupied segregated fields. The amount of cortical territory related to the aphakic, optically corrected eye depended on the manipulations of the fellow eye. In competition with an unmanipulated fellow eye, the aphakic eye's territory was greatly reduced. In competition with a part-time occluded eye, its territory was reduced to a lesser degree, depending on the duration of the occlusion. In competition with a continuously occluded eye, however, the space related to the aphakic, optically corrected eye was slightly greater than that related to the occluded eye. Since neither the aphakic nor the continuously occluded eye receives normal visual input, they are both impaired. Therefore, they may compete on an almost equal basis for synaptic territory in layer 4C of striate cortex. Moreover, it is likely that activities originating in the aphakic and the continuously occluded eye are asynchronous, and that this condition is sufficient to drive the postnatal segregation of inputs from the two deprived eyes.


Expression of GABA and GABAA receptors by neurons of the subplate zone in developing primate occipital cortex: evidence for transient local circuits. Several lines of evidence suggest that the transient subplate zone of the embryonic mammalian telencephalon could influence cortical development through synaptic or trophic interactions with growing cortical afferents and migrating neurons. Since such interactions may involve neurotransmitters and their receptor molecules, we have examined the expression of GABA and subunits of the GABAA/benzodiazepine receptor complex in the occipital lobe of embryonic rhesus monkeys by immunochemistry and in situ hybridization. We found that during the second half of gestation, when the subplate zone reaches peak maturity in this species, many neurons can be immunolabeled with both GABA antisera and monoclonal antibodies against GABAA receptor subunits. The most robust labeling occurs at approximately embryonic day (E)125 (birth is at E165). Electron microscopic observations of receptor subunit-immunolabeled material confirmed that subunits of the GABAA receptor are localized in the subplate neurons and their dendritic processes. In many instances the reaction product is associated with the plasma membranes of labeled processes, some of which form symmetrical synapses with small unlabeled axon terminals. The results of in situ hybridization are in accord with the results of receptor subunit immunochemistry. From E80 to E141, hybridization signal for GABAA receptor subunit mRNA occurs in the subplate zone and increases steadily to peak levels between E125 and E141. The present results reveal that all the elements necessary for the formation of functional GABAergic synaptic circuitry are present in the subplate zone. Further, the ages showing the most pronounced receptor and transmitter expression in this primate coincide with the ingrowth of major cortical afferent systems. Taken together, these findings suggest that GABAergic local neuronal circuits in the subplate may be involved in the development of long tract connections stationed in this zone prior to their transfer to the overlying cortical plate.


Effect of a two-solution fluoride mouthrinse on remineralization of enamel lesions in vitro. A previous study showed that a two-solution fluoride (F) rinse deposited significantly more loosely-bound F on the tooth surface than did a sodium fluoride (NaF) rinse with the same F concentration (12 mmol/L). In the present study, this experimental rinse was evaluated for its ability to cause remineralization of enamel lesions in an in vitro pH-cycling model. Caries-like lesions were formed in the enamel of extracted human molars by means of a pH 4 demineralizing solution. Fifty-one approximately 120-microns-thick sections containing lesions were randomly divided into (1) control, (2) NaF rinse, and (3) two-solution F rinse groups. With the cut surfaces protected, the control samples were immersed in a pH 7 remineralizing solution for 12 days, and twice daily the sections were also exposed to a pH 4 demineralizing solution for 30 min. Samples in the NaF group received an additional one-minute rinse with a NaF (12 mmol/L) solution twice daily. Samples in the two-solution rinse group received the rinse treatment with a 12 mmol/L F solution prepared by combination of a Na2SiF6 and phosphate-containing solution with a calcium solution just before use. The mineral contents of the lesions were assessed by quantitative microradiography. The results showed that (1) no significant de- or remineralization was detected in the controls; (2) a 46% decrease in mineral loss (delta Z) of the lesion was produced by the NaF rinses; and (3) a 94% decrease in delta Z and a 20-microns-thick, mineral-dense surface-coating were produced by the two-solution F rinse treatment.


Silica-induced precipitation of calcium phosphate in the presence of inhibitors of hydroxyapatite formation. The promotion and the inhibition of hydroxyapatite formation by various substances were determined by measurement of the induction time of spontaneous precipitation (ti) from supersaturated solutions. Silica was found to decrease ti in Hepes-buffered (pH 7.2) supersaturated solutions with a wide range of calcium-to-phosphate ratios and concentrations. Also, in suspensions of the oral bacteria S. mutans or C. matruchotii in 1 mmol/L calcium, 7.5 mmol/L phosphate, and 50 mmol/L Hepes (pH 7.2), silica was capable of stimulating precipitation. Macromolecules derived from these bacteria by freezing and thawing appeared to be strong inhibitors of calcium phosphate precipitation. In the presence of silica, the effects of these bacterial inhibitors could be partially overcome, which supports the idea that silica in dental plaque is a promoter of calculus formation. In contrast, inhibition of calcium phosphate precipitation by a low-molecular-weight inhibitor, pyrophosphate, could not be counteracted by silica.


Risk level, knowledge, and preventive behavior for human papillomaviruses among sexually active college women. During the past decade, there has been mounting scientific evidence linking human papillomavirus (HPV) with cervical cancer and, at the same time, a great increase in physician consultations for HPV infections. HPV infection risk factors include multiple sex partners, early age at first intercourse, history of sexually transmitted diseases (STDs), and smoking. This study surveyed 263 sexually active college women and concluded that (1) they are at considerable risk for contracting HPV, (2) they lack awareness of HPV, and (3) they are not practicing preventive behaviors that could reduce their risk of HPV and its serious consequences.


Selective impairment of neuroendocrine and hemodynamic responses to a mu-opioid peptide in aged rats. The objective of this study was to determine if there are age-related alterations in hemodynamic and/or neuroendocrine responses to the mu-opioid receptor agonist, [D-Ala2,MePhe4,Gly(ol)5] enkephalin (DAMGO), or corticotropin releasing hormone (CRH) administered centrally. To this end, DAMGO (1-3 nmoles) or CRH (1 nmole) was injected intracerebroventricularly (icv) to freely moving young (6-8 month) and aged (24-26 month) Fischer 344 male rats. Blood pressure, heart rate (HR), and plasma concentrations of norepinephrine (NE), epinephrine (EPI), adrenocorticotropin (ACTH), and prolactin (PRL) were measured over time. Under basal conditions, NE levels were higher and blood pressures were lower in aged rats, whereas there were no significant differences in EPI, ACTH, or PRL levels. The stimulatory effect of DAMGO on blood pressure, HR, and plasma EPI and ACTH was attenuated, but the PRL response was enhanced in aged cohorts. In contrast, there were no age-related differences in the NE responses to DAMGO or CRH nor in CRH-induced increases in EPI or ACTH. The sympathoadrenal and hemodynamic effects of DAMGO were blocked by naloxone in both age groups. These results indicate that alterations in mu-opioid function with age are specific for the opioid system and do not reflect a generalized decline in central regulation of neuroendocrine and cardiovascular function.


[Celioscopic removal of cysts: a fortuitous histological discovery of an ovarian carcinoma. A case report]. The authors report a case of cancer of the ovary that had not been recognised but which was discovered after laparoscopic surgical cystectomy. Laparoscopy is very reliable in a presumptive diagnosis of the nature of an ovarian mass, but in this case shows that the operator has to be very attentive especially when there is a rapid recurrence of cyst formation in the ovary that has been operated on. They discuss the indications for intra-peritoneal laparoscopic ovariectomy in short term recurrences after ovarian cyst removal and in particular when these were mucinous. For most authors the risk of dissemination when an ovarian cyst which has proved to be malignant is punctured, seems to be nil, and unlikely to effect the prognosis. It may be possible that in the future the diagnosis of these cases can be improved when the clinical picture, the ultrasound picture and the laparoscopic picture are added to a new development of intraperitoneal laparoscopic ultrasound which will make it possible to view the contents of the cyst with direct contact.


Myogenes and myotubes.

Myogenes and myotubes. Investigation of the congenital myopathies has been limited by a lack of knowledge regarding basic mechanisms involved in normal myogenesis of human muscle and the relative rarity of these diseases. A newly recognized family of regulatory genes has been shown to be necessary for myogenesis to proceed to formation of normal mature muscle. It is likely that investigation of patients with one or more types of structural myopathy may show that abnormalities of the regulatory basic helix-loop-helix (bHLH) genes may be responsible for disease.


Enhanced synthesis of gelatinase and stromelysin by myoepithelial cells during involution of the rat mammary gland. During the involution of the mammary gland there is destruction of the basement membrane as the secretory alveolar structures degenerate. Immunofluorescence staining of sections of rat mammary gland with antibodies to 72 KD gelatinase (MMP-2) and stromelysin (MMP-3) revealed increased production of these two proteinases during involution. This increased expression was mostly restricted to myoepithelial cells. Increased expression during involution was also demonstrated by immunoblotting techniques. Gelatin zymography indicated that the predominant metalloproteinase present in involuting rat mammary glands was a 66 KD gelatinase.


Subtalar joint arthroereisis with SILASTIC silicone sphere: a retrospective study. Many authors have discussed the various surgical techniques of subtalar joint arthroereisis in the treatment of flexible flatfoot. Some authors advocate the use of bone grafts in the sinus tarsi to limit excessive subtalar joint pronation, while others advocate the use of endoprosthetic devices. These authors will present a retrospective study of subtalar joint arthroereisis utilizing a SILASTIC silicone sphere technique devised by the senior authors.


Tolerance induction in resting memory B cells specific for a protein antigen. Resting memory B lymphocytes specific for the model protein Ag cytochrome c have been shown to be susceptible to tolerance induction in in vitro splenic fragment cultures. This induction of nonresponsiveness is dependent upon the strength of the interaction between surface Ig and specific Ag, where concentration, valency, affinity, and time of exposure all appear to be important factors, as is the case for tolerance induction in immature or primary B cells. The induction of nonresponsivenes in greater than 80% of Ag-specific memory B cells was achieved by incubation with 1 microM cytochrome polymer for 24 h in the absence of T cell help. Not only were memory B cells unresponsive to specific Ag, they were also unable to become activated through nonspecific uptake and presentation of an Ag to which T cells have been primed, demonstrating that the induction of nonresponsiveness involves more than a modulation or blockade of surface Ig receptors. Although soluble factors collected from activated T cells failed to prevent memory B cells from becoming nonresponsive after surface Ig cross-linking, the direct activation of T cells within splenic fragment cultures did partially inhibit tolerance induction in splenic fragment memory B cells. In addition, the induction of tolerance was partially blocked by protein tyrosine kinase inhibitors, suggesting a physiologic change within the B cells associated with the state of nonresponsiveness and resulting from tyrosine-specific phosphorylation.


Anti-tumor necrosis factor antibody therapy fails to prevent lethality after cecal ligation and puncture or endotoxemia. Cytokines have been studied intensively to delineate their role in the altered pathophysiology observed in septic shock. We studied the role of TNF in the lethality of two well characterized models of septic shock by inhibiting TNF's activity with a specific antibody. In the first model, sepsis was induced by cecal ligation and puncture (CLP), and in the second model sepsis was induced by either an i.p. or i.v. injection of LPS. After CLP, plasma endotoxin was detectable within 4 h and reached a peak at 8 h (136 +/- 109 ng/ml). TNF bioactivity peaked at 12 h (528 +/- 267 pg/ml) at a significantly higher level than sham-operated control mice (64 +/- 31 pg/ml). After i.p. LPS, TNF peaked much more quickly (90 min) compared with CLP and at a significantly higher level (107,900 +/- 25,000 pg/ml). Another cytokine studied in septic shock, IL-6, peaked at 12 h after CLP at 1011 +/- 431 pg/ml, and at 90 min after lethal LPS at 16,300 +/- 3,700 pg/ml. Mice were treated with an anti-TNF antibody that has been shown previously to inhibit in vivo TNF activity. Antibody treatment of mice subjected to CLP significantly reduced TNF bioactivity but did not reduce mortality or pulmonary neutrophilic infiltration. In the i.v. LPS model, anti-TNF antibody treatment concomitant with LPS injection reduced plasma TNF activity from 80,000 +/- 20,000 pg/ml to undetectable levels. However, anti-TNF treatment immediately before either i.v. or i.p. LPS did not reduce mortality. Additionally, when the antibody was administered 4 h before the lethal i.v. LPS, there was no reduction in lethality. These data show that in two separate models of septic shock blockade of TNF biologic activity will not prevent lethality.


IFN-gamma mediates increased glucocorticoid receptor expression in murine macrophages. Exposure of the murine macrophage cell line, RAW 264.7, to murine rIFN-gamma resulted in a significant increase in the number of glucocorticoid receptors (GcR). A doubling in the number of GcR was observed as early as 24 h after rIFN-gamma treatment, and receptor number was maximal by 36 h after rIFN-gamma treatment and represented approximately a fourfold increase. Scatchard analysis indicated that a twofold increase in GcR affinity was concomitant with the rIFN-gamma-induced increase in GcR number in RAW 264.7 cells. Increased GcR numbers were induced after exposure of RAW 264.7 cells to as little as 0.1 U/ml rIFN-gamma, and optimal expression was observed at 5 U/ml. Treatment of peritoneal exudate macrophages from C3H/OuJ mice and the LPS hyporesponsive mouse strain, C3H/HeJ, with rIFN-gamma induced an approximately twofold increase in the GcR with no concomitant change in receptor affinity. These results suggest that IFN-gamma may be essential not only for macrophage activation, but also for increasing macrophage sensitivity to feedback inhibition by glucocorticoids by increasing the number and/or affinity of available GcR.


Effects of eleven cytokines and of IL-1 and tumor necrosis factor inhibitors in a human B cell assay. The effects of different recombinant human cytokines and cytokine inhibitors were compared in a culture system in which cell contact with mutant EL-4 thymoma cells of murine origin efficiently stimulates human B cell proliferation and Ig secretion in conjunction with human T cell supernatant. IL-1 alpha, IL-1 beta, TNF-alpha, and IL-2 co-stimulated B cell proliferation and IgM, IgG, and IgA secretion, whereas IL-3, IL-4, IL-5, IL-6, IFN-gamma, or GM-CSF had weak or no activity in this regard. In contrast, TGF-beta 1 was strongly inhibitory. A very strict hierarchy of cytokine interactions was found in that IL-1 was necessary to induce TNF-alpha responsiveness, and TNF-alpha the IL-2 responsiveness, of the B cells. Most likely the small number of starting B cells in the present assay (300 FACS-separated B cells/200 microliters) minimized the effects of autocrine B cell factors. IL-4 together with IL-1 induced IgE secretion, and the IgE secretion was further increased by TNF-alpha. IFN-gamma had no modulatory effect on the IL-4 dependent IgE response in this system. Pretreatment of B cells with IL-1R antagonist (IL-1ra, which binds to IL-1R) or addition of soluble TNF receptor type 1 (sTNF-R55, which binds to TNF) completely inhibited the IL-1 or TNF-alpha effects, respectively. This occurred in a specific manner; the inhibition was reversed by a large excess of cytokine. IL-1ra also inhibited a B cell response induced by PMA-preactivated EL-4 cells alone. Because B cells responding to such preactivated EL-4 cells did not acquire TNF-alpha responsiveness, no IL-1 was apparently involved under this assay condition. It appears, therefore, 1) that IL-1ra can act on B cells and 2) that this antagonist may not only block IL-1R, but may provide a direct or indirect inhibitory signal interfering even with IL-1-independent B cell activation.


Preferential involvement of a phospholipase A2-dependent pathway in CD69-mediated platelet activation. CD69 is a signal transducing disulfide-linked homodimer functionally expressed on platelets, CD3bright thymocytes, and activated lymphocytes. In an attempt to investigate early molecular events in CD69-mediated cell activation we studied the relative contribution of phospholipase A2 (PLA2) and phosphatidylinositol-specific phospholipase C-dependent pathways during platelet activation induced by CD69 stimulation. Thromboxane A2 (TXA2) synthetase inhibitor and TXA2R inhibitor R68070 were able to inhibit platelet aggregation induced by CD69 stimulation, indicating that TXA2 was the main mediator of the response. CD69-induced arachidonic acid release and TXA2 production were essentially PLA2 dependent because they could be blocked by the PLA2 inhibitor quinacrine. Inositol 1,3,4-trisphosphate generation was clearly detectable after CD69 cross-linking, but it was completely abrogated by quinacrine and R68070 and therefore secondary to TXA2 release and TXA2R engagement. Finally, direct measurement of enzymatic activity in vitro using radiolabeled phospholipid vesicles showed that CD69 cross-linking resulted in PLA2-dependent arachidonic acid and lysophosphatidylcholine generation from phosphatidylcholine, which was sensitive to quinacrine but not to R68070. By contrast, CD69-induced 1,2-diacylglycerol release from phosphatidylinositol 4,5-bisphosphate was blocked by both inhibitors. These results indicate a preferential involvement of PLA2 in CD69-dependent signal transduction in platelets and provide evidence for the unique role of PLA2-mediated activation pathways in transmembrane receptor signaling.


Inhibition of basal and tumor necrosis factor-enhanced binding of murine tumor cells to murine endothelium by transforming growth factor-beta 1. The adherence of cells to microvascular endothelium is important in a number of processes, including inflammatory responses and metastasis. It has been demonstrated that in human models, cytokines such as TNF, IL-1, IFN-gamma increase the adhesiveness of endothelium for cells of the immune and inflammatory system by stimulating the expression of cell adhesion molecules on endothelial cell surfaces. We and others have shown similar cytokine-induced endothelial adhesiveness for tumor cells in murine and human models. In contrast to the effect of those modulators, transforming growth factor-beta (TGF-beta) has been shown to inhibit the binding of human neutrophils and T lymphocytes to human endothelium, although the mechanism of TGF-beta action remains unknown. Little is known about the effect of TGF-beta on tumor cell-endothelial interaction. In the present study, we demonstrate that TGF-beta inhibits basal and TNF-enhanced binding of murine P815 mastocytoma cells to murine microvascular endothelium (MME). The alterations in MME mediated by TGF-beta, also lead to the inhibition of adherence of murine splenocytes, thymocytes, and human lymphoblastoid cells but do not inhibit adherence of murine B16 melanoma cells. The effect of TGF-beta is transient and inhibition of the endothelial adhesive phenotype is strongest 12 to 24 h after addition of the factor to MME. The TGF-beta-mediated inhibition of P815 basal binding to endothelium is dependent on protein synthesis because cycloheximide reverses the TGF-beta effect. TGF-beta does not appear to activate classical signal transduction pathways. Inhibitors of G proteins do not abolish TGF-beta action, protein kinase C and protein kinase A activators elicit an effect opposite to that of the factor, TGF-beta does not increase intracellular cAMP levels, and finally calcium-mobilizing agents do not mimic, but rather inhibit the effect of TGF-beta. However, TGF-beta-mediated inhibition of both basal binding and TNF-enhanced P815 binding to MME is completely abolished in the presence of the protein phosphatase inhibitor okadaic acid which suggests that TGF-beta may elicit its effect by stimulating protein phosphatase activity.


Interferon pretreatment regulates interferon and interleukin-6 production in L929 cells in a coordinated manner. Two sublines of L929 cells with different interferon (IFN)-producing capacities synthesized IFN and interleukin-6 (IL-6) simultaneously in response to Sendai virus infection. IFN pretreatment primed the production of both cytokines. However, the difference in IL-6 production between the "high and low producer" L-cell sublines was about one magnitude of order larger than in the case of their IFN production. The determination of the corresponding mRNA levels also reflected this difference.


Occurrence and characterization of acyclovir-resistant herpes simplex virus isolates: report on a two-year sensitivity screening survey. For the past 2 years, a survey network was established for the screening of acyclovir (ACV)-resistant clinical isolates of herpes simplex virus (HSV). Among 889 strains tested for in vitro ACV sensitivity, 14 HSV-1 and 6 HSV-2 were resistant to ACV concentrations exceeding 3 micrograms/ml. These resistant isolates were most often obtained after prolonged ACV treatment of severely immunocompromised patients. For five patients, the emergence of ACV-resistant virus correlated with treatment failure. In particular, a decrease in the in vitro sensitivity to ACV was observed for eight successive HSV-1 isolates from one immunodeficient patient undergoing therapy. All ACV-resistant isolates were studied for their sensitivity to different antiherpetic compounds and showed various cross-sensitive and -resistant patterns. The examination of viral populations by plaque autoradiography procedures frequently revealed their heterogeneity in terms of thymidine kinase (TK) phenotype and allowed the detection of various proportions of TK-positive (TK+), TK-deficient (TKD), or TK-altered (TKA) viruses. Our data underline the importance of monitoring the emergence of drug-resistant virus during the course of antiviral therapy, and the need for the detection and characterization of TK mutants in clinical specimens. The routine examination of drug sensitivity of HSV isolates provides useful information to clinicians for the management of ACV treatment in the hope of preventing ACV-resistant mutants from becoming predominant in mixed viral populations.


Hepatitis C virus RNA in saliva of patients with posttransfusion hepatitis and low efficiency of transmission among spouses. In a prospective study of posttransfusion hepatitis, 14 patients who were diagnosed with posttransfusion hepatitis C were enrolled randomly for the study of hepatitis C virus (HCV) RNA in saliva. Saliva and serum samples were collected on the same day. Spouses of 11 married patients were also tested for anti-C100 and HCV RNA. Paired serum and saliva samples were tested for HCV RNA by a nested polymerase chain reaction (PCR). Two primer pairs specific for the non-coding region of HCV were used for the PCR and a oligonucleotide sequence between the primers was used as the probe for Southern hybridization. Six patients were positive for HCV RNA by first round PCR amplification and an additional four patients were detected after second round PCR. All patients were negative for HCV RNA in saliva after first round PCR, while seven were positive after second round PCR amplification. All seven patients were positive for HCV RNA in paired serum samples. HCV RNA was detectable in saliva from 1 week to 38 months after the onset of hepatitis. All spouses were negative for anti-C100 and HCV RNA. We conclude that HCV RNA is present in the saliva of approximately half of patients with acute and chronic hepatitis C, and the presence of HCV RNA correlates with HCV viremia. The efficiency of HCV transmission is low among spouses.


Correlation between cyclophosphamide-induced viral susceptibility and depletion of Junin virus-induced suppressor populations. In contrast to lymphocytic choriomeningitis virus, another arenavirus, Junin virus (JV), the etiologic agent of Argentine hemorrhagic fever, when inoculated into suckling mice, induces lethal meningoencephalitis characterized by a delayed-type hypersensitivity (DTH)-like immune response. However, the adult BALB/c mouse is resistant to infection and no DTH reaction can be seen. This different viral sensitivity may be related to the development of an antigen non-specific DTH-suppressor cell pathway at work in the adult mouse. When the resistant mice are treated with cyclophosphamide (Cy) (50 mg/kg each dose) given at days -1,+1,+4 (zero: infection day), animals become susceptible and develop DTH reaction in brain that leads to death. We analyze the influence of the timing of Cy administration on the suppressor system developing after infection. It was found that Cy depletes the previously described JV-induced suppressor populations (Tsv) but a new suppressor cell (Tsv*) is disclosed bearing the Thy 1+ Ly1+2- phenotype which is unable to depress DTH in Cy-treated animals. With only two doses of Cy corresponding to days -1 and +1, the target of Tsv* cells is depleted but the third dose is still required to achieve full depletion of Tsv cells which are able to employ the Cy-resistant antigen-specific suppressor cells as targets. Since the Cy treatment is able to deplete the Tsv population together with the target of Tsv* cells, animals became unable to regulate lethal DTH reaction. Thus, a cellular explanation for an empirically established Cy schedule able to abrogate the adult mouse resistance to JV is proposed.


Platinum levels and clinical responses of tumours treated by cisplatin with and without concurrent hyperthermia: a case study. The heterogeneity of platinum distributed within tissue after clinical administration of cisplatin was evaluated for the first time. Platinum levels were correlated with the observed clinical responses of separate superficial histiocytic sarcomas located in the forearm of a 74-year-old male patient. One of four lesions received four weekly treatments with hyperthermia administered concurrently with 30 mg/m2 cisplatin, while three lesions were treated with cisplatin alone. The lesion receiving hyperthermia concurrently with cisplatin had a solid partial response during a 6-week period following this therapy, whereas two other tumours receiving cisplatin alone progressed. One lesion could not be clinically evaluated. Platinum levels were determined in multiple samplings from three of the four lesions and normal tissue in order to evaluate the validity of taking a single tumour sample of 100 mg or less for the analysis of platinum content. Such a small single sample might provide a value significantly different from the true average because of sampling error. The range in platinum distribution encompassed an average of three-fold difference within eight separate sample groups, with a factor of six being the greatest difference in a single sample group. This degree of heterogeneity is great enough to suggest that conclusions made from the analysis of small and single random tissue samples could be sufficiently in error to misdirect investigative or medical decisions.


Temperature enhancement of syncytium formation by HIV and Sendai virus. Syncytium formation, the characteristic cytopathic effect (CPE) of the human immunodeficiency virus (HIV) and cell fusion by Sendai virus, is accelerated by increasing the ambient temperature to values at which normal metabolic activity is inhibited. Uninfected C8166, CEM, and H9 cells were absorbed at 4 degrees C onto monolayers of H9 cells chronically infected with HIV and incubated subsequently at either 37 degrees C or 45 degrees C. Similarly chick and human erythrocytes and Hela cells were agglutinated with Sendai virus at 4 degrees C before incubation at temperatures of up to 50 degrees C. With both viruses the rate of cell fusion was directly related to temperature. Since membrane fluidity is dependent on the phase-transition temperature points of the membrane lipids it is proposed that sufficient membrane fluidity is essential for cell fusion to occur. The implication of these observations on the cytopathology of HIV is discussed.


Peripheral neuropathy and myonecrosis following hyperthermia and radiation therapy for recurrent prostatic cancer: correlation of damage with predicted SAR pattern. During the past 10 years, numerous phase I-II studies were conducted and provided clinical experience with combined radiation therapy and hyperthermia treatments. Among the rare complications reported in these combined radiation therapy-hyperthermia trials were myonecrosis and peripheral neuropathy which were felt, mainly on a clinical basis, to be caused by local heat damage. Recently, such complications were noted in two patients with recurrent prostatic cancer treated in our department with radiation therapy combined with deep regional hyperthermia delivered by the Sigma-60 applicator of the BSD 2000 hyperthermia system (Salt Lake City, UT, USA). Analysis of the results of three-dimensional modelling of the SAR (specific absorption rate, W/kg) pattern in these patients indicated high SAR at the sites of the complications. Pretreatment three-dimensional modelling or other methods of predicting potential areas of high power deposition may have a role for future hyperthermia treatment planning aiding in the prevention of possible local heat damage and providing improved delivery of heat to the target volume.


Transfusion-acquired cytomegalovirus infection in children in a hyperendemic area. Thirty-nine children without previous cytomegalovirus (CMV) infection received blood transfusion in the National Taiwan University Hospital. The overall transfusion-acquired CMV infection rate was 36% (14/39). Donor CMV seropositive rate was 70%. None of the nine children who had received seronegative blood became infected, in contrast to 14 of the 21 children (67%) who had received seropositive blood (P = 0.002). Another significant risk factor associated with CMV infection was the use of fresh blood: 13 of 15 (87%) with fresh seropositive blood were infected, in contrast to one of six (17%) with "old" seropositive blood (P = 0.01). Most of the fresh blood was used within 24 hours. This blood processing method was shown to account for the extremely high rate of CMV infection in those who had received fresh seropositive blood. The results indicated that the incidence of CMV infection can be reduced by avoiding the use of fresh blood, especially blood less than 24 hours old. For such a population in Taiwan with high prevalence of positive CMV antibody, this approach was more applicable than screening donor blood for CMV antibody.


Detection of HPV DNA in archival specimens of cervical cancer using in situ hybridisation and the polymerase chain reaction. An archival survey of 98 cervical cancer specimens dating from the 1920s to the 1980s was undertaken to determine whether changes had occurred in the prevalence of human papilloma-virus (HPV) DNA. HPV DNA was detected in paraffin sections of cancers fixed in 10% formalin by in situ hybridisation (ISH) using HPV 6, 11, 16, and 18 32P-labelled DNA probes under conditions of high stringency; and by the polymerase chain reaction (PCR) using 20-mer oligonucleotide primers to amplify 109 bases of the E6 region of HPV 16. In 30 instances results obtained from Southern blot hybridisations which had been carried out on specimens of fresh tissue from the same cancers collected during the 1980s were available for comparison. The rates of HPV DNA detection in cervical cancers ranged from 83% (by Southern or PCR) and 70% (by ISH) on specimens from the 1980s, to 50% and 63% (by ISH and PCR, respectively) on specimens from the 1920s. HPV 16 was by far the most common type, being identified by Southern or ISH in approximately 92% of HPV DNA-positive specimens. No significant change in the prevalence of HPV DNA, or of HPV types, in cervical cancers was found over the 65 year period examined.


Immunoglobulin G subclasses and lymphocyte stimulatory responses to cytomegalovirus in transplant patients with primary cytomegalovirus infections. The early activation of T- and B-cell responses to cytomegalovirus was studied in immunosuppressed patients. Primary lymphocyte stimulation to cytomegalovirus (CMV) antigen, a measure of T-helper activity, and anti-CMV IgG subclass responses were analyzed. Ten patients suffering from primary CMV infection following renal transplantation were studied. Of the ten, nine became positive for CMV induced lymphocyte proliferation 5-40 weeks after transplantation. Nine showed an almost simultaneous appearance of anti-CMV IgG1 and three at 3-32 weeks after transplantation, while one patient synthesized only low levels of anti-CMV IgG1. The lymphocyte proliferation assays have limited diagnostic value for primary CMV infection in renal transplant patients. The humoral and cellular immune responses seemed to be independent of each other.


Synthesis of glial fibrillary acidic protein in rat C6 glioma in chemically defined medium: cyclic AMP-dependent transcriptional and translational regulation. Glial fibrillary acidic protein (GFA) expression was induced in rat C6 glioma in chemically defined medium by the addition of N6, O2'-dibutyryl cyclic AMP (dbcAMP). Induction was dependent on the increase in intracellular cyclic AMP (cAMP), which was linearly correlated with added dbcAMP. Contrary to GFA mRNA synthesis, which can be obtained by cAMP-dependent and -independent pathways, translation of mRNA into GFA was observed only above a cellular cAMP concentration of approximately 0.2 fmol/cell. dbcAMP stimulation did not affect the vimentin concentration, which remained at a low level, but changed the cellular morphology from a bipolar to a stellate shape. A similar morphological change was observed after stimulation of C6 with lipopolysaccharide (LPS). However, LPS did not significantly increase the intracellular concentration of cAMP and the LPS-induced mRNA was not translated into GFA. Our results indicate that GFA synthesis is regulated at the mRNA level and at the translational level and that a cAMP-dependent mechanism determines the ultimate synthesis of GFA by a yet unknown mechanism.


Membrane localization of endopeptidase-24.11 and peptidyl dipeptidase A (angiotensin converting enzyme) in the pig brain: a study using subcellular fractionation and electron microscopic immunocytochemistry. Brains from piglets were dissected and a block of tissue including the substantia nigra, globus pallidus, and entopeduncular nucleus was homogenized and then fractionated on discontinuous Percoll gradients. Ligand-binding assays using (-)-[3H]nicotine and [3H]quinuclidinyl benzilate served to delineate fractions containing nicotinic and muscarinic acetylcholine receptors. In this system endopeptidase-24.11 exhibited a biphasic distribution, consistent with its presence on both pre- and postsynaptic membranes. Peptidyl dipeptidase A (angiotensin converting enzyme; ACE) was associated with membrane fractions containing muscarinic receptors. An immunoblot of these fractions with an affinity-purified polyclonal antibody to ACE revealed only the neuronal form of ACE (Mr 170,000), the endothelial form (Mr 180,000) being undetectable. Electron microscopic immunoperoxidase staining of the substantia nigra, with an affinity-purified antibody to endopeptidase-24.11 at the preembedding stage, showed this antigen to be confined to the plasma membranes of boutons, axons, and some dendrites. Both pre- and postsynaptic membranes were stained, and occasionally other regions of the dendritic membrane were positive. No staining of synaptic vesicles within the boutons was observed. Thus, two independent approaches indicate that endopeptidase-24.11 is present on both pre- and postsynaptic membranes in the pig substantia nigra. The subcellular fractionation suggests that neuronal ACE is confined to dendritic membranes.


Purification and characterization of naturally occurring benzodiazepine receptor ligands in rat and human brain. Chemicals that are active at the benzodiazepine receptor (endozepines) are naturally present in the CNS. These substances are present in tissue from humans and animals and in plants and fungi. Using selective extraction protocols, HPLC purification, receptor binding displacement studies, and selective anti-benzodiazepine antibodies, we have identified six or seven peaks of endozepines in rat and human brain. All material could competitively displace [3H]flunitrazepam binding to cerebellar benzodiazepine binding sites. Two peaks also competitively displaced Ro 5-4864 binding to the mitochondrial benzodiazepine binding site. Total amounts of brain endozepines were estimated to be present in potentially physiological concentrations, based on their ability to displace [3H]flunitrazepam binding. Although endozepine peaks 1 and 2 had HPLC retention profiles similar to those of nordiazepam and diazepam, respectively, gas chromatography-mass spectrometry as well as high-performance TLC revealed biologically insignificant amounts of diazepam (less than 0.02 pg/g) and nordiazepam (less than 0.02 pg/g) in the purified material. Electrophysiologically, some purified endozepines positively modulated gamma-aminobutyric acid (GABA) action on Cl- conductance, monitored in patch-clamped cultured cortical neurons or in mammalian cells transfected with cDNA encoding various GABAA receptor subunits. These studies demonstrate that mammalian brains contain endozepines that could serve as potent endogenous positive allosteric modulators of GABAA receptors.


Concerted CMP-dependent [3H]inositol labeling of phosphoinositides and agonist activation of phospholipase C in rat brain cortical membranes. [3H]Inositol ([3H]Ins) labeling of phosphoinositides was studied in rat brain cortical membranes. [3H]Ins was incorporated into a common lipid pool through both CMP-dependent and independent mechanisms. These are as follows: (1) a reverse reaction catalyzed by phosphatidyl-inositol (PtdIns) synthase, and (2) the reaction performed by the PtdIns headgroup exchange enzyme, respectively. Membrane phosphoinositides prelabeled in either CMP-dependent or independent fashions were hydrolyzed by guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S)- and carbachol-stimulated phospholipase C. Unlike CMP-dependent labeling, however, CMP-independent incorporation of [3H]Ins into lipids was inhibited by 1 mM (0.04%) sodium deoxycholate. Thus, when PtdIns labeling and phospholipase C stimulation were studied in a concerted fashion, [3H]Ins was incorporated into lipids primarily through the PtdIns synthase-catalyzed reaction because of the presence of deoxycholate required to observe carbachol-stimulation of phospholipase C. Little direct breakdown of [3H]PtdIns was detected because production of myo-[3H]inositol 1-monophosphate was minimal and myo-[3H]inositol 1,4-bisphosphate was the predominant product. Although PtdIns labeling and 3H-polyphosphoinositide formation were unaffected by GTP gamma S and carbachol and had no or little lag period, GTP gamma S- and carbachol-stimulated appearance of 3H-Ins phosphates exhibited an appreciable lag (10 min). Also, flux of label from [3H]Ins to 3H-Ins phosphates was restricted to a narrow range of free calcium concentrations (10-300 nM). These results show the concerted activities of PtdIns synthase, PtdIns 4-kinase, and phospholipase C, and constitute a simple assay for guanine nucleotide-dependent agonist stimulation of phospholipase C in a brain membrane system using [3H]Ins as labeled precursor.


gamma-Aminobutyric acidA receptor pharmacology in rat cerebral cortical synaptoneurosomes. Equilibrium binding interactions at the gamma-aminobutyric acid (GABA) and benzodiazepine recognition sites on the GABAA receptor-Cl- ionophore complex were studied using a vesicular synaptoneurosome (microsacs) preparation of rat brain in a physiological HEPES buffer similar to that applied successfully in recent GABAergic 36Cl- flux measurements. NO 328, a GABA reuptake inhibitor, was included in the binding assays to prevent the uptake of [3H]muscimol. Under these conditions, the equilibrium dissociation constant (KD) values for [3H]muscimol and [3H]diazepam bindings are 1.9 microM and 40 nM, respectively. Binding affinities for these and other GABA and benzodiazepine agonists and antagonists correlate well with the known physiological doses required to elicit functional activity. This new in vitro binding protocol coupled with 36Cl- flux studies should prove to be of value in reassessing the pharmacology of the GABAA receptor complex in a more physiological environment.


Alterations of benzodiazepine receptor binding in tremor rats with absence-like seizures. Tremor rats begin to exhibit clinical or electrical absence-like seizures after 6 weeks of age, and by 14 weeks of age, all have seizures. Central-type benzodiazepine receptor binding was investigated in tremor rats and control rats, aged 4 weeks and 16 weeks. Significantly lower benzodiazepine receptor density and no differences in affinity were found in the hippocampus of the tremor rats in comparison with that of control rats at both ages. This abnormality is considered to be due to a tremor gene and may be the cause of absence-like seizures in tremor rats. A significantly lower receptor density was found in the cerebellum at 4 weeks of age in the tremor rats than in the control rats. These changes may be related to tremorous movements in the tremor rats. Receptor density was significantly lower in the brainstems of tremor rats and control rats at 16 weeks of age than at 4 weeks of age, and the decrease was more marked in control rats. These facts may reflect a reduced decrease in the response to the dysfunction of gamma-aminobutyric acidergic neurons, or the function of the gamma-aminobutyric acid/benzodiazepine receptor system may be secondarily increased to suppress seizures in 16-week-old tremor rats.


Modulation of extracellular gamma-aminobutyric acid in the ventral pallidum using in vivo microdialysis. Intracranial microdialysis was used to investigate the origin of extracellular gamma-aminobutyric acid (GABA) in the ventral pallidum. Changes in basal GABA levels in response to membrane depolarizers, ion-channel blockers, and receptor agonists were determined. Antagonism of Ca2+ fluxes with high Mg2+ in a Ca(2+)-free perfusion buffer decreased GABA levels by up to 30%. Inhibition of voltage-dependent Na+ channels by the addition of tetrodotoxin also significantly decreased basal extracellular GABA concentrations by up to 45%, and blockade of Ca2+ and Na+ channels with verapamil reduced extracellular GABA by as much as 30%. The addition of either the GABAA agonist, muscimol, or the GABAB agonist, baclofen, produced a 40% reduction in extracellular GABA. GABA release was stimulated by high K+ and the addition of veratridine to increase Na+ influx. High K(+)-induced release was predominantly Ca(2+)-dependent, whereas the effect of veratridine was potentiated in the absence of extracellular Ca2+. Both high K(+)- and veratridine-induced elevations in extracellular GABA were inhibited by baclofen, whereas only veratridine-induced release was antagonized by muscimol. These results demonstrate that at least 50% of basal extracellular GABA in the ventral pallidum is derived from Ca(2+)- or Na(+)-dependent mechanisms. They also suggest that Na(+)-dependent release of GABA via reversal of the uptake carrier can be shown in vivo.


Phosphatidylinositol kinase is reduced in Alzheimer's disease. Phosphatidylinositol (PI) kinase and PI phosphate (PIP) kinase activities were measured in postmortem samples of brain tissue from patients with Alzheimer's disease and nondemented control subjects. A membrane-free cytosolic fraction from four neocortical locations, with exogenous inositol lipids as the substrate, was used. Tissue from patients with Alzheimer's disease was characterized by reduced PIP formation; the reduction was 50% in prefrontal cortex, temporal cortex, and parietal cortex and 40% in precentral gyrus. In contrast, no alterations were found in PI bisphosphate formation in these four neocortical locations. The specific changes in PI kinase but not PIP kinase activity suggest that the findings may have functional relevance to the involvement of brain membrane processes in Alzheimer's disease.


Lymphoma-induced polyradiculopathy in AIDS: two cases. Progressive polyradiculopathy is a rare, well-documented complication of the acquired immunodeficiency syndrome in man. It has been commonly attributed to a cytomegalovirus (CMV) infection. We report two HIV-infected patients with clinical and electrophysiological features of a unique, subacute, progressive polyradiculopathy. Post-mortem examination in case 1 disclosed an infiltration of the leptomeninges, the lumbar spinal cord, and the anterior and posterior roots by a B-cell immunoblastic lymphoma. Immunochemistry for HIV1 and CMV was negative in the peripheral and the central nervous system. Case 2 showed bone-marrow involvement by a Burkitt type lymphoma. Specific chemotherapy was followed by both clinical improvement of the polyradiculopathy and complete remission on a second bone-marrow biopsy. These findings may indicate that a lymphoma must also be considered a possible cause of polyradiculopathy in AIDS.


The course of neuropathy after cessation of cisplatin treatment, combined with Org 2766 or placebo. Peripheral neuropathy is an important and disabling side-effect of cisplatin treatment. A new drug, Org 2766, has been found to prevent this neuropathy up to 1 month after treatment. A group of 18 patients with ovarian cancer, who participated in an earlier randomized study with placebo or Org 2766, together with cisplatin and cyclophophamide, were thereafter prospectively followed up to 2 years after discontinuation of treatment to monitor the development of neurological signs and symptoms and vibration perception threshold (VPT). Exploratory, descriptive data analysis shows that between 1 and 4 months after the last cycle the average sum score for neurological signs and symptoms and VPT had deteriorated compared with 1 month after treatment. Thereafter a gradual but incomplete improvement was seen between 4-12 and 12-24 months after treatment. These changes were seen in all patients regardless of previous treatment with Org 2766 or placebo, but deterioration was less pronounced in patients previously treated with Org 2766. These results suggests that treatment with Org 2766 to prevent a cisplatin-induced neuropathy should possibly be continued up to 4 months after the last cycle of cisplatin.


Sequential quantitative scintigraphy of parotid glands with chronic inflammatory diseases. A practical, time-saving procedure for sequential quantitative scintigraphy is introduced and 4 parameters chosen from 12 parameters by discriminant analysis are used to evaluate the function of the parotid gland. The examination was performed in 120 cases, including 16 cases with recurrent parotitis in childhood, 33 with chronic obstructive parotitis (COP), 37 with Sjögren's syndrome (SS), 4 with sialadenosis, and 30 normal controls. The scintigraphic findings were analyzed and compared with the histologic findings. The diagnostic value of this method was investigated and scaling for differential diagnosis of COP and SS was established. Scintigraphy is considered to be a useful method for evaluation of parotid function and as a diagnostic aid for SS and COP, especially in patients in whom sialography cannot be performed.


Idiopathic trigeminal sensory neuropathy. Idiopathic trigeminal sensory neuropathy is a rare disorder characterized by transient sensory disturbances in the territory of one or more branches of the trigeminal nerve. Acute and chronic forms of the disease have been recognized, but the boundaries between these groups are not clearly defined. Six cases of idiopathic trigeminal sensory neuropathy are reported. All patients presented with sensory abnormalities limited to the territory of one trigeminal nerve, in the absence of other clinical features. The symptoms were on the left side of the face in five cases and on the right side in one. No bilateral involvement was seen. There were no cases of muscle weakness, and the corneal reflex was always present. In two cases, sensory disturbances began in the tongue or lips and later spread through the entire territory of the three trigeminal branches. Both patients recovered completely after 3 months. The other four patients had involvement of the second and third branches (two cases), or the third branch alone (two cases), and the symptoms persisted unmodified for 2 to 4 years. There were no clinical variables at onset to enable one to predict an acute or chronic evolution of the disease.


OKT3 treatment of steroid-resistant rejection in pediatric liver transplant recipients. Of a total of 187 consecutive liver grafts in 149 pediatric recipients, 59 episodes of steroid-resistant, biopsy-proven rejection (32% of grafts) were treated with OKT3 monoclonal antibody. After 59 OKT3-treated episodes, liver function at the end of treatment was normal in 40%, improved in 35%, and unchanged in 24%. Of 21 partial responses, 12 episodes eventually resolved to yield an overall complete response rate of 59%. CD3-positive T-cells greater than 5% occurred during 61% of OKT3-treated rejection episodes and was associated with impaired efficacy of OKT3 (30% complete response rate). Six grafts were re-treated with OKT3 for rejection and in all CD3-positive T-cells could not be maintained less than 5%. Of the six grafts requiring repeat OKT3, five failed-retreatment and required retransplantation. OKT3 antibodies of low titer were found prior to a second use of OKT3 in 65% of episodes. Patients treated with OKT3 after failing more than two preceding steroid courses had a significantly increased chance of graft loss (57%; p = 0.01). We conclude that this group of pediatric patients appeared less responsive to OKT3 compared to other series combining pediatric and adult recipients, possibly due to a more vigorous immune response in the child.


The relationship between cyclic adenosine 3',5'-monophosphate and morphology in exponential phase Candida albicans. The relationship between changes in cyclic AMP content and germ tube formation in exponential phase Candida albicans was investigated using two simple media containing glucose plus ammonium chloride, or N-acetyl-D-glucosamine (GlcNAc). The glucose medium did not promote germ tube formation unless the cells were starved before inoculation, whereas the GlcNAc medium promoted germ tube formation in both non-starved and starved cells. The cyclic AMP content of exponential phase cells, non-starved cells and starved cells was 0.21, 0.34 and 0.64 pmol mg-1 dry wt, respectively. In glucose medium, cyclic AMP content in both non-starved cells and starved cells increased for a period of 60 min after inoculation, but then decreased for a further 120 min. The cyclic AMP content of non-starved cells and starved cells was 0.16 and 0.29 pmol mg-1 dry wt, respectively, after 180 min. The maximum percentage of non-starved cells with germ tubes was around 20%. Starved cells with germ tubes were observed after 40 min and reached a maximum (around 90%) after 140 min. The number of germ tubes remained constant for the next 40 min. In GlcNAc medium, the cyclic AMP content of both non-starved cells and starved cells showed a tendency to increase for 180 min. The content of non-starved cells and starved cells was 2.02 and 1.75 pmol mg-1 dry wt, respectively, after 180 min. Germ tube formation in non-starved cells started after 70 min, reached around 80% after 150 min, and remained stable for the next 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)


Treponema denticola and Porphyromonas gingivalis as prognostic markers following periodontal treatment. Subgingival plaque samples were collected from individuals with advanced periodontitis before and 3 to 11 weeks after scaling and root planing periodontal treatment. The plaque levels of Treponema denticola and Porphyromonas gingivalis antigens were measured before and after treatment by a quantitative immunoassay procedure using monoclonal antibodies specific for these oral bacteria. A decrease in mean levels of T. denticola (P less than .05) and P. gingivalis antigens (P less than .09) were observed following periodontal therapy. Improved health, as measured by a decrease in probing depth, was associated with a decrease in T. denticola antigen (P less than .05). These results suggest that the T. denticola levels of successfully treated sites decreased, while non-responding sites had levels of this microbial marker which were equal to or greater than the pre-treatment levels. These results provide additional evidence that T. denticola is associated with human adult severe periodontal disease, and can serve as a prognostic marker for disease recurrence.


Microbiology of healthy and diseased periodontal sites in poorly controlled insulin dependent diabetics. A group of poorly-controlled insulin dependent diabetes mellitus (IDDM) patients were examined in a cross-sectional design for total microbial levels, microbial incidence, and the percent levels of selected periodontal microorganisms. These organisms were selected on the basis of prior reports that associated them with either periodontal disease or health. One periodontally-healthy and one periodontally-diseased site were examined in each IDDM patient. Increased levels of the periodontal pathogens Prevotella intermedia, P. melaninogenica spp., Bacteroides gracilis, Eikenella corrodens, Fusobacterium nucleatum and Campylobacter rectus (formerly Wolinella recta) were found at the periodontal diseased sites. Increased prevalence of the organisms P. intermedia, P. melaninogenica spp., and C. rectus were found at the diseased sites. A significantly higher percentage of P. intermedia was found at the sites exhibiting deep pockets and attachment loss.


Apoptosis: a gene-directed programme of cell death. Apoptosis is a particular type of programmed cell death which commonly occurs in the developing embryo, in normal healthy adult tissues and in many pathological settings. In contrast to necrosis, apoptosis is not a passive phenomenon but is gene-directed, usually requiring ongoing protein synthesis. The dying cell is characterised by having a raised level of cytosolic Ca2+; this activates a non-lysosomal Ca(2+)- and Mg(2+)-dependent endonuclease which digests the chromatin into oligonucleosome length fragments. The dying cell may or may not fragment into a number of apoptotic bodies, but in all cases the cell contents are bounded by a membrane which prevents the spillage of harmful substances such as DNA. Apoptotic cells are eliminated through phagocytosis by neighbouring cells and macrophages, and cell surface changes on apoptotic cells aid their recognition and engulfment by the phagocytosing cells. Extrinsic signals can both stimulate and inhibit apoptosis, and even direct damage to the cell can activate the process. Apoptosis is widely involved in organ formation in the embryo, and its occurrence in response to noxious stimuli such as cytotoxic drugs, irradiation and hyperthermia may be viewed as an altruistic suicide. Apoptosis provides a safe disposal mechanism for neutrophils at inflamed sites, and within the immune system it is considered responsible for eliminating self-reactive T-cell clones and for the affinity maturation of antibody producing cells. A failure to undergo apoptosis has been invoked in the pathogenesis of low-grade follicular lymphoma, and the triggering of apoptosis with monoclonal antibodies specifically in tumour cells has been achieved in one or two cases.


A criterion based audit of inpatient asthma care. Closing the feedback loop. We have assessed the care of patients admitted to a specialist respiratory medical ward acutely ill with asthma, using a criterion based audit derived from a standard management protocol already in use in our hospitals. The audit was first performed from 01.01.90 to 31.08.90; after implementing certain changes, the audit was repeated from 1.12.90 to 31.1.91. Special attention was paid in each audit review to pre-admission measures, inpatient management and pre-discharge and follow-up management. During both audit periods, of a total of 78 patients, 74 patients gave a reason for the worsening of their asthma; 59 had had PEF measured and 58 had received systemic steroids before admission; 77 patients had full objective assessment of severity on admission; 76 patients were discharged on oral steroids; 62 had PEF meters for home monitoring; and 65 of the 68 patients who lived in our district were seen again within six weeks as outpatients in the chest clinic. However, only 30/55 (54%) had PEF variability of 20% or less (our criterion for appropriateness of discharge, in the first audit period) and only 32/55 had a written check on their inhaler technique in the first audit period. By relaxing our PEF criterion for discharge (in line with national guidelines), by introducing a stamp for recording that inhaler technique had been checked, and with encouragement and exhortation from senior staff, we improved our performance of meeting the set standards to 17 of 23 (74%) patients for PEF variability and to 22 of 23 (96%) patients for written check on inhaler technique in the second audit period.(ABSTRACT TRUNCATED AT 250 WORDS)


Lunate Silastic arthroplasty in Kienbock's disease. The use of Silastic lunate replacement for Kienbock's disease has many advocates. We reviewed the long-term results of eight patients who had this procedure performed. Although good short-term results were obtained, this was not maintained and only two patients were regarded as satisfactory at an average follow-up time of 44 months. Evidence of synovitis was apparent in seven cases. It is now our view that if surgical intervention is necessary for this condition, the best surgical option is probably a scapho-trapezio-trapezoid fusion.


Pyrimido[1,6-a]benzimidazoles: a new class of DNA gyrase inhibitors. Substituted 4-quinolone- (1, A = CH) and 1,8-naphthyrid-4-one- (1, A = N) 3-carboxylic acids are currently the only classes of clinically useful antibacterial agents exerting their activity by inhibiting the subunit A of DNA gyrase. Pyrimido[1,6-alpha]benzimidazoles 11 have been found to be a new class of inhibitors of this enzyme. The design, synthesis, and biological activity of these compounds are reported.


Synthesis and antiviral activity of acyclic nucleosides with a 3(S),5-dihydroxypentyl or 4(R)-methoxy-3(S),5-dihydroxypentyl side chain. Optically pure acyclic nucleoside analogues with a 3(S),5-dihydroxypentyl or 4(R)-methoxy-3(S),5-dihydroxypentyl side chain were synthesized starting from 2-deoxy-D-ribose. The acyclic nucleosides were obtained by alkylation of the bases with the mesylates 16 and 17. Of these series of novel nucleoside analogues only 9-[3(S),5-dihydroxypent-1-yl]guanine (6d) showed marked antiviral activity. It inhibited the cytopathogenicity of herpes simplex virus type 1 (HSV-1) at a concentration of 0.4-0.6 microgram/mL, which thus points to a greater antiviral activity than recently reported for the mixture of the R and S enantiomers (12.5 micrograms/mL). In contrast with 6d, its 4(R)-methoxy derivative 7d did not show antiviral activity, which implies that the 4'-methoxy group is unable to mimic the 1',4'-oxygen bridge of the normal furanose ring.


Visceral leishmaniasis in infancy and childhood epidemiology and clinicopathological study of 63 cases in Al-Baha Province, Saudi Arabia. The epidemiology, clinicopathological features, and response to therapy of 63 Saudi patients with visceral leishmaniasis are described. The clinical features in our cases were similar to those described from Asir province, India, and Ethiopia, except for the presence of lymphadenopathy. Fever, hepatosplenomegaly, pancytopenia, and liver dysfunction were common findings. The unusual feature is the seasonal variation in the distribution of the disease. The response to sodium stibogluconate was excellent and the mortality rate was low (less than 1 per cent).


Clinical aspects of kala-azar in children from the Sudan: a comparison with the disease in adults. The clinical presentation of kala-azar in 43 children and 45 adults was compared. In both groups fever, left upper quadrant abdominal pain and swelling, and weight loss were equally the most common presenting symptoms. Lymphadenopathy was observed in 86 per cent of children and 76 per cent of adults. Splenomegaly was absent in 2 per cent of children and 7 per cent of adults. No significant difference was found in frequency distribution of symptoms and signs between children and adults. Haematological indices were compared in both children and adults with kala-azar and their control groups. In both children and adults with kala-azar, haemoglobin concentration, total white cell count, and platelet count were significantly lower before than after treatment. Only haemoglobin concentration was lower in children with kala-azar as compared with adults with the disease. Children in the control group had lower haemoglobin and higher total white cell count than adult controls. Response to therapy was evaluated in 693 patients. Two-hundred-and-fifty children and 373 adults were treated with sodium stibogluconate 10 mg/kg for 30 days; in both groups 12 per cent deaths and 4 per cent relapses occurred. Thirty children and 40 adults were treated with sodium stibogluconate 2 x 10 mg/kg for 15 days. In children, 3 per cent deaths and 7 per cent relapses were noted; in adults there were 8 per cent deaths and 5 per cent relapses. No significant difference in death rate or relapse rate was found between children and adults in both regimens. Both regimens performed equally well in children and adults with regard to death rate and relapse rate.


Pharmacokinetics of enrofloxacin in fingerling rainbow trout (Oncorhynchus mykiss). The pharmacokinetics of intravenously and orally administered enrofloxacin was determined in fingerling rainbow trout (Oncorhynchus mykiss). Doses of 5 or 10 mg enrofloxacin/kg body weight were administered intravenously to 26 fish for each dose and blood was sampled over a 60-h period at 15 degrees C. Two groups of fish were treated orally with 5, 10, or 50 mg/kg (80 fish/dose at each temperature) and held at 15 degrees C or 10 degrees C during the 60-h sampling period. Following intravenous administration, the serum concentration-time data of enrofloxacin in rainbow trout were best described by a two-compartment open model for both doses of 5 and 10 mg enrofloxacin/kg. The hybrid rate constants alpha and beta did not differ between doses. The distributional phase was rapid with a half-life of 6-7 min for both doses. Overall half-lives of elimination were 24.4 h (95% CI = 20.2-30.8) and 30.4 h (24.2-41.0), respectively, for the 5- and 10-mg/kg doses. A large Vd(area) was observed following dosing of either 5 or 10 mg enrofloxacin/kg,: 3.22 and 2.56 l/kg, respectively. Whole body clearance for 5 mg/kg was 92 ml/h.kg and 58 ml/h.kg at the 10-mg/kg dose. Following oral administration, the serum concentration-time data for enrofloxacin were best described as a one-compartment open model with first-order absorption and elimination. Apparent Ka over all doses at 10 degrees C averaged 62% less than apparent Ka at 15 degrees C. Estimates of the apparent t(1/2)e over both temperatures ranged from 29.5 h (18.4-73.4) to 56.3 h (38.3-106.6). Bioavailability averaged 42% over all doses at 15 degrees C and was decreased to an average of 25% at 10 degrees C. Peak serum concentrations appeared between 6 and 8 h following dosing. A dose of 5 mg/kg/day was estimated to provide average steady-state serum concentrations at 10 degrees C that are approximately 4.5 times the highest reported MIC values for Streptococcus spp., the fish pathogen least sensitive to enrofloxacin. Owing to the long apparent half-life of elimination of enrofloxacin in fingerling trout, it would take approximately 5 to 9 days to achieve these predicted steady-state serum concentrations; this estimate is important when considering the duration of therapy in clinical trials.


Effect of systemic L-arginine administration on hemodynamics and nitric oxide release in man. The effects of L-arginine administration on systemic hemodynamics and plasma concentrations of neuro-endocrine hormones and amino acids were investigated in 10 normotensive healthy volunteers. Nitrite/nitrate in urine and cyclic guanosine monophosphate (c-GMP) in plasma were also measured as indicators of release of nitric oxide (NO). L-arginine administration (30 g/300 ml/30 min) caused hypotension (mean arterial pressure; 79.3 +/- 3.9 mmHg fell to 68.8 +/- 2.2 mmHg) with tachycardia (62.3 +/- 2.3 beats/min increased to 67.5 +/- 1.9 beats/min). The plasma concentration of L-arginine before administration was 98.8 +/- 8.2 mumol/l and increased to 7263 +/- 567 mumol/l 20 min after administration. Cardiac output also increased to 127.2 +/- 3.9% by L-arginine administration. Total peripheral resistance was calculated to fall to 65.9 +/- 2.0%. L-arginine administration slightly changed several hormones, but all values were within normal ranges. Nitrite/nitrate in urine increased 142.1 +/- 12.4% compared to the values before L-arginine administration. Plasma concentrations of c-GMP and L-citrulline, the by-product of NO from L-arginine, were also significantly increased by L-arginine administration. All our results provide evidence for the first time that systemically administered L-arginine releases NO in man.


Rapid reconstruction of three dimensional images from ultrasonography using a linearly moving mirror. A system for reconstructing three dimensional (3-D) images from ultrasonography using a linearly moving mirror was modified. The device could display the 3-D structure of objects in real time by employing an on-line system. In a case of hepatocellular carcinoma, structures of the tumor and blood vessels were selectively visualized. One 3-D image was generated within five minutes. This modified system will be of use in the analysis of liver tumors.


Immunohistochemical detection of cluster 1 small cell lung cancer antigen and chromogranin A in lung carcinomas. Eighteen small cell lung cancers (SCLCs), 108 non-SCLCs (67 adenocarcinomas, 29 squamous cell carcinomas and 12 large cell carcinomas) were immunohistochemically examined for expressions of cluster 1 SCLC antigen/N-CAM and chromogranin A with monoclonal antibodies NCC-Lu-243 and anti-chromogranin A. The cell membranes of all the SCLCs and three of the 67 adenocarcinomas (4.5%) were stained for cluster 1 SCLC antigen/N-CAM. Eight of the 18 SCLCs (44.4%), and three of the 67 adenocarcinomas (4.5%) were stained for chromogranin A, but no squamous cell carcinoma or large cell carcinoma was stained for both antigens. Two of the three adenocarcinomas which expressed cluster 1 SCLC antigen/N-CAM had been suspected of being either SCLC or poorly-differentiated adenocarcinoma cytologically, and were resected after chemotherapy and radiotherapy. Histologically, they were poorly-differentiated adenocarcinoma with rosette-like tubules. The remaining one was moderately-differentiated papillary adenocarcinoma resembling bronchial surface epithelial cells without mucin (BSE type adenocarcinoma). The three adenocarcinomas which expressed chromogranin A were well- to moderately-differentiated BSE type adenocarcinomas, and stained tumor cells were distributed sparsely as neuroendocrine cells in the normal bronchial mucosa. One of them also expressed cluster 1 SCLC antigen/N-CAM. In the present study, we demonstrated the usefulness of NCC-Lu-243 in the immunohistochemical detection of adenocarcinomas with neuroendocrine features.


Evidence for bovine leukemia virus infection of peripheral blood monocytes and limited antigen expression in bovine lymphoid tissue. Bovine leukemia virus (BLV) infection of cattle is a common but inapparent retrovirus infection from which less than 5% of infected cattle manifest clinically with lymphoma. During the course of life-long infection, cattle maintain a high anti-BLV titer during an apparent latent infection of B cells from which proviral DNA but not transcripts can be detected. To investigate if BLV infection is truly latent and restricted only to B lymphocytes, peripheral blood leukocytes were purified from cattle with naturally acquired BLV infection with various stages of subclinical and clinical disease. These cells were purified into populations of polymorphonuclear cells, monocytes and subsequently B cells and T cells by fluorescent-activated cell sorting. Southern blot analysis revealed the presence of provirus in monocytes and B cells but not T cells. Secondly, the ability of provirus containing cell types to express BLV antigens was then confirmed by mitogen stimulation in vitro. Subsequently, the pattern of expression of BLV antigens was studied in situ in tissue sections to determine the location of BLV antigen expressing cells in vivo. Antigen expressing cells were infrequent and solitary in intrafollicular and marginal zone cells of architecturally normal lymph nodes of chronic, BLV-infected cattle. These results demonstrate that BLV persists in cells of the monocyte/macrophage lineage in addition to B lymphocytes and that expression of BLV antigens in cattle doses occur but rarely in lymph nodes of BLV infected cattle. The high frequency of provirus containing cells and infrequent expression of BLV antigen in vivo suggests that BLV expression is not truly latent but highly regulated, possibly triggered by rare events in host lymphoid tissue.


Expression of retinoic acid receptor genes in developing rat livers and hepatoma cells. The expression of retinoic acid receptor alpha, beta, and gamma mRNA was examined in developing rat livers and rat hepatoma-derived cell lines H-4-II-E, McA-RH 7777, and 8994 that represent different hepatocyte phenotypes. Northern blot hybridization demonstrated that all three receptor mRNAs were expressed in the fetal livers of different gestational ages, and the levels of expression increased significantly 3 to 4 weeks after birth. In the hepatoma cell lines, the expression pattern of retinoic acid receptor alpha and gamma mRNA did not correlate with the phenotype. In contrast, retinoic acid receptor beta mRNA was only detected in the adult phenotypic H-4-II-E cells but not in McA-RH 7777 and 8994 cells, which represent embryonic and fetal hepatocyte phenotypes, respectively. The levels of retinoic acid receptor beta mRNA in hepatoma cell lines were lower than adult rat liver. These data suggest that the increased expression of retinoic acid receptor beta gene is associated with differentiation or maturation of rat hepatocytes. The effect of retinoic acid on retinoic acid receptor gene expression was also studied in hepatoma cells. Retinoic acid did not regulate retinoic acid receptor gene expression in McA-RH 7777 and 8994 cells, and the retinoic acid receptor beta gene remained inactivated in these cells. However, Southern blot hybridization indicated that the gross structure of retinoic acid receptor beta gene was not altered during malignant transformation. In H-4-II-E cells, retinoic acid increased the expression of retinoic acid receptor beta and gamma gene. Because of the similarity between H-4-II-E cells and normal adult hepatocytes, this type of autoregulation may be a mechanism by which retinoic acid regulates its own effect in vivo.


Bone metastases of hepatocellular carcinoma after liver resection. Between January 1985 and July 1990, 323 cases of hepatocellular carcinoma underwent liver resection in our department. Bone metastases were found in 12 of these cases (3.7%). Bone metastases were mainly found in vertebral bone (58.3%) and pelvic bone (41.7%). The time interval to the development of bone metastasis after liver resection was closely related to the presence of intrahepatic metastasis and the stage at operation. In all cases, the initial clinical symptom was pain and/or motor disturbance. Radiotherapy was performed in 10 cases and transcatheter arterial embolization or surgery was performed in 4 cases. The pain or neurological symptoms improved with these therapies in all cases. Cumulative survival was 1 year in 74%, 2 years in 34%, and 3 years in 17%, respectively.


Primary hepatic tumours in children: a 26-year review. Twenty-one children were admitted to a single paediatric institution between 1964-1990 with histologically proven primary liver tumours. The diagnosis was hepatoblastoma (HBL) in 15 patients, hepatocellular carcinoma (HCA) in 2, rhabdomyosarcoma (RMS) in 2, non-Hodgkin's lymphoma (NHL) in 1, and haemangioendothelioma (HE) in 1. The common presenting clinical features were anaemia, abdominal mass, and abdominal pain. Serum alpha-foetoprotein was useful in establishing a diagnosis in HBL and in monitoring disease activity. Computed tomographic (CT) scan, ultrasound, and angiography were useful preoperative investigations for assessing site and resectability of tumour. There were no survivors in patients with malignant hepatic tumours (n = 10) who had surgery alone prior to 1981. Of 7 patients with HBL diagnosed after 1981 who had adequate surgical resection and chemotherapy, 5(72%) are currently alive and disease free between 15 months and 8 years from diagnosis. We conclude that adequate surgical resection and adjuvant chemotherapy can improve disease free survival for children with HBL. Optimal treatment has yet to be devised for other malignant hepatic tumours.


The feasibility of hind foot amputation in selected sarcomas of the foot. The treatment of foot sarcomas is generally a below knee amputation. In selected sarcomas of the forefoot, however, a transtarsal amputation according to Chopart, a calcaneotibial arthrodesis according to Pirogoff, or a supramalleolar amputation according to Syme can be considered the treatment of choice, leading to an optimal functional outcome.


Epstein-Barr virus detection in neck metastases by polymerase chain reaction. Cervical nodal metastasis from occult carcinomas represents a diagnostic challenge. This is a common presentation of undifferentiated nasopharyngeal carcinoma (UNPC), but metastatic carcinomas from other sites must be considered. UNPC has the distinguishing feature of a close association with Epstein-Barr virus (EBV). Since the polymerase chain reaction (PCR) can detect EBV in archival tissues, it offers significant advantages over previous methods for the detection of viral genomes. Its extreme sensitivity allows analysis of small samples from needle aspirates. Using the polymerase chain reaction to amplify EBV sequences from archival tissues, 15 of 18 NPC samples were positive for EBV. Of these 18, 14 of 14 with UNPC were positive, 1 of 2 with squamous cell carcinoma (SCC) were positive, and 10 of 2 with adenocarcinoma were positive. All 6 UNPC metastatic to lymph nodes were positive. Carcinoma metastatic to cervical nodes from 17 of 17 non-UNPC occult primaries lacked EBV. This demonstrates the utility of EBV detection by the polymerase chain reaction in the evaluation of patients with metastases to neck nodes from occult primary carcinomas in order to identify cases of UNPC.


Effect of coolant flow on the performance of round sapphire tips in a saline field. Round sapphire and silica tips have been studied as surgical probes for focusing Nd:YAG laser radiation during various surgical indications. Since most of these data have been obtained with silica tips from Surgical Laser Technology, there are limited data on the physical performance of round sapphire tips. An investigation of round sapphire tip performance during tissue ablation was undertaken. Nd:YAG laser power was delivered in repetitive 2-sec exposures to a round sapphire tip placed on fresh skinned chicken breast. Total exposures of 1,000 J were performed at different saline perfusion rates. The tip's performance in terms of tissue perforation rate, blanched tissue area, tissue heating rate, and forward transmission was measured. Best tip performance occurred at the lowest cooling rates and was independent of the tip's forward transmission value. No physical deterioration of the tip's optical surface was observed. Round sapphire tip performance in a saline field is primarily determined by the tip's temperature.


The effects of lithium isotopes on the myo-inositol 1-phosphatase reaction in rat brain, liver, and testes. Enzyme inhibition studies were performed with several lithium isotopes in order to more precisely define how lithium inhibits the enzyme myo-inositol 1-phosphatase. This lithium-induced inhibition is thought to be central to the therapeutic effects of lithium in the treatment of manic-depressive disorder. Naturally occurring lithium (NLi) exists as a combination of isotopes: 6Li and 7Li. Lethality studies were performed comparing 6LiCl, 7LiCl, and NLiCl, did not demonstrate a differential effect as previous studies had suggested. Enzyme inhibition studies were performed with these individual lithium isotopes, and compared to the effects of the naturally occurring combination (NLi) on the inhibition of myo-inositol 1-phosphatase using a partially purified enzyme preparation from rat brain, liver and testes. Identical inhibition was observed with all lithium isotopes and their combinations. In addition, both D- and L-myo-inositol 1-phosphates were used as enzyme substrates and found to be equivalent. These experiments, along with previous work demonstrating lithium acting as an uncompetitive inhibitor in the reaction, and the lack of lithium binding sites on the enzyme, suggests the hypothesis that lithium is possibly inhibiting this reaction by interfering with the formation of a transition cyclic intermediate, myo-inositol 1,3-cyclic phosphate, which may be formed from either the D- or L-substrates. This proposal is in contrast to previous suggestions regarding the inhibitory mechanism of action of lithium on the myo-inositol 1-phosphatase reaction.


[Detection of herpes simplex virus 2 (HSV) IgG and IgM using ELISA in transsexuals and homosexuals]. In this study, the sera of 12 cases of transsexuals with vaginoplasty and 15 cases of homosexuals have been investigated with ELISA from the view point of HSV-2 IgG and IgM. Twenty-five cases were within the 18-26 age groups and the other 2 transsexuals were 36 years old. Our study group has not significant clinical pathology. All of the 27 cases of transsexuals and homosexuals have been found positive for HSV-2 IgG. In 7 cases HSV-2 IgM have been found positive, the other 20 cases gave negative results. In the one third of the transsexuals and one fifth of the homosexuals, HSV-2 IgG have been found positive. Twenty-six percent of seropositivity in transsexuals and homosexuals show that they are the significant risk group for HSV-2 infection.


Primary human herpesvirus 6 infection in young children. Human herpesvirus 6 (HHV-6) is a recently discovered virus that, on the basis of serologic evidence, appears to infect most children by the age of three years. However, the clinical manifestations of primary HHV-6 infection have not been well defined. We studied consecutive children two years old or younger who presented to an emergency ward with febrile illnesses. Our evaluation included the isolation of HHV-6 from peripheral-blood mononuclear cells, an immunofluorescent-antibody assay, the detection of HHV-6 by the polymerase chain reaction (PCR), and restriction-endonuclease-fragment profiles of HHV-6 isolates. HHV-6 was isolated from 34 of 243 acutely ill children (14 percent). The children with viremia had irritability, high temperatures (mean, 39.7 degrees C), and inflammation of tympanic membranes (in 21), but few other localizing signs. Two children were hospitalized, but all 34 recovered after an average of four days of fever. The rash characteristic of roseola, which has been associated with HHV-6 infection, was noted in only three children. In 29 children (85 percent), serum samples obtained during convalescence had at least a fourfold increase in IgG antibody titers; 4 infants less than three months old who presumably had maternal antibody did not have this increase. HHV-6 was isolated from blood obtained during convalescence in only one child, but in two thirds of the children the virus could be detected by PCR. The isolates had genomic heterogeneity, indicating the presence of multiple strains. Primary infection with HHV-6 is a major cause of acute febrile illness in young children. Such infection is associated with varied clinical manifestations, viremia, and the frequent persistence of the viral genome in mononuclear cells.


Specific low-affinity recognition of major histocompatibility complex plus peptide by soluble T-cell receptor. The T-cell receptor is necessary and sufficient for recognition of peptides presented by major histocompatibility complex molecules. Other adhesion molecules, like CD4 or CD8, play an auxiliary role in antigen recognition by T cells. Here we analyse T-cell receptor (TCR) binding using a soluble rather than a cell-bound receptor molecule. A TCR-immunoglobulin chimaera is constructed with the variable and the first constant regions of both the TCR alpha- and beta-chains linked to the immunoglobulin light-chain constant regions. This soluble TCR is expressed, assembled and secreted as an alpha beta heterodimer by a myeloma cell line transfected with the recombinant genes. Furthermore, the soluble TCR is biologically active: it specifically inhibits antigen-dependent activation of the relevant T-cell clones and thus discriminates between proper and irrelevant peptides presented by major histocompatibility complex molecules.


A protein of the endoplasmic reticulum involved early in polypeptide translocation. To identify components of the mammalian endoplasmic reticulum involved in the translocation of secretory proteins, crosslinking and reconstitution methods were combined. A multispanning abundant membrane glycoprotein was found which is in proximity to nascent chains early in translocation. In reconstituted proteoliposomes, this protein is stimulatory or required for the translocation of secretory proteins.


Calcium-dependent enhancement of calcium current in smooth muscle by calmodulin-dependent protein kinase II. Calcium entry through voltage-activated Ca2+ channels is important in regulating many cellular functions. Activation of these channels in many cell types results in feedback regulation of channel activity. Mechanisms linking Ca2+ channel activity with its downregulation have been described, but little is known of the events responsible for the enhancement of Ca2+ current that in many cells follows Ca2+ channel activation and an increase in cytoplasmic Ca2+ concentration. Here we investigate how this positive feedback is achieved in single smooth muscle cells. We find that in these cells voltage-activated calcium current is persistently but reversibly enhanced after periods of activation. This persistent enhancement of the Ca2+ current is mediated by activation of calmodulin-dependent protein kinase II because it is blocked when either the rise in cytoplasmic Ca2+ is inhibited or activation of calmodulin-dependent protein kinase II is prevented by specific peptide inhibitors of calcium-calmodulin or calmodulin-dependent protein kinase II itself. This mechanism may be important in different forms of Ca2+ current potentiation, such as those that depend on prior Ca2+ channel activation or are a result of agonist-induced release of Ca2+ from internal stores.


[Experiences with arthrotomy of the knee in day care]. A retrospective study was performed of patients who had undergone an open knee procedure treated as day cases at the Regional Hospital of Zevenaar. An analysis was made of the rate of complications, whether complications were treated adequately, and how patients experienced an arthrotomy of the knee as a day-case procedure. Eighty-seven healthy patients aged 15 to 50 years underwent an arthrotomy of one knee as a day-case procedure, secondary to a diagnostic arthroscopy. The mean follow-up period was 8 months. Patients were reviewed by questionnaire, after which the knee was examined. The number of complications (6) was comparable with those reported in the literature. Complications within a few days after operation were reported to the GPs, who either treated themselves, or referred patients to the hospital. Of the patients 76% preferred this procedure to an operation in a clinical setting. The amount of pain suffered appears to be of great influence on the preference of the patient. The results of this study demonstrate that an arthrotomy of the knee is feasible as a day-care procedure, although extensive pre- and postoperative instructions to the patients may be improved.


[The physiopathological aspects of the atrial natriuretic factor]. The original observation by de Bold et al. (1981) of a rapid, massive, and short-lasting diuretic and natriuretic effect following injection of rat atrial extracts into intact rats, led to the identification, isolation and purification of the atrial natriuretic factor (ANF). ANF is stored in atrial myocytes and released into the blood stream by atrial distension. Available data suggest that the mechanism of ANF-induced natriuresis involves either renal hemodynamic effects, such as the increase in glomerular filtration rate and reduction of medullary tonicity, or direct effect on sodium transport in the medullary collecting ducts. ANF induces relaxation of vascular smooth muscle, decreases blood pressure and cardiac output. All these effects displayed by ANF are associated to the an inhibition of aldosterone, renin and vasopressin release. Most of these actions are mediated by specific high affinity receptors, which are coupled to a particulate guanylate cyclase. Although ANF levels are increased in some disorders, such as severe heart failure, hypertension, chronic renal failure, the role of the peptide is uncertain. To better define the potential physiopathological role and the possible therapeutic implications of this new hormonal system in conditions of disturbed body fluid and sodium homeostasis, further experimental and clinical data must be awaited.


Mitoxantrone in patients affected by hepatocellular carcinoma with unfavorable prognostic factors. Patients affected by hepatocellular carcinoma (HCC) with unfavorable prognostic factors have limited therapeutic options due to moderate responsiveness to chemotherapeutic agents and lack of compliance with such treatments. In this study the feasibility and efficacy were evaluated of a treatment with mitoxantrone(dihydroxyanthracenedione, DHAD), 12 mg/m2 i.v. on day 1 every 3 weeks. We included 18 patients with poor-risk HCC ineligible for our other trials in relation to their performance status [8], pretreatments [6], age over 70 years [5], severe concomitant illness [6], and altered blood count [platelets less than 100,000/mm3) [8]. Of 17 evaluable cases, there were 4 partial remissions (23%) (95% confidence interval, 10-47%), 6 no changes, and 7 progressions. There were no drug-related deaths, and side effects were moderate and documented only in a few cases. Median survival of evaluable patients was 5 months (range, 1-15). The results were comparable with those obtained with adriamycin but without important toxicity. We conclude that DHAD seems to be safe and moderately active in poor-risk HCC.


Human papillomavirus type 16 DNA in oral white sponge nevus. White sponge nevus (WSN) is a benign hereditary lesion of the mucous membranes. DNA extracted from a biopsy specimen of oral WSN was assayed for the presence of DNA sequences homologous to human papillomavirus (HPV) types 1, 2, 4, 6, 11, 13, 16, and 18 by Southern blot hybridization. Only HPV-16 homologous DNA sequences were detected at a copy number of approximately 200 to 250 genome copies per diploid cell. The viral DNA sequences did not appear to be integrated into the host cell chromosome. The finding of HPV-16 in an inherited lesion such as WSN indicates that caution must be exercised in ascribing a causal association in relation to the demonstration of HPV in other mucosal disorders.


Adsorption of a hydrophobic mutagen to dietary fiber from taro (Colocasia esculenta), an important food plant of the South Pacific. The incidence of colorectal cancer is lower in Polynesian populations of the South Pacific than in European populations. This difference in incidence of the disease may be, at least partly, related to diet. Dietary fiber is believed to protect against colorectal cancer, and one of the ways it may act is by adsorbing mutagens that are carcinogenic. Very little is known about the chemical composition or the ability to adsorb mutagens of these dietary fibers from South Pacific food plants. In contrast to European food plants, which are mostly dicotyledons, South Pacific food plants are mainly monocotyledons. We isolated cell walls (dietary fiber) from the three edible parts of taro (Colocasia esculenta), which is a monocotyledon and a major South Pacific food plant. The ability of these three unlignified cell-wall preparations to adsorb the hydrophobic environmental mutagen 1,8-dinitropyrene was studied. The greatest adsorption occurred with walls from leaf blade, followed by petiole and corm walls, although the differences were not major. The amount of adsorption was intermediate between the low adsorption previously found with unlignified dicotyledon walls (from the flesh of potato tubers and immature cabbage leaves) and the much higher adsorption found with unlignified walls from monocotyledons of the grass and cereal family (Poaceae) (from leaves of seedling Italian ryegrass). These data are consistent with the monosaccharide compositions of the taro wall preparations, which were more similar to those of unlignified walls of dicotyledons than to unlignified walls of the Poaceae. These findings are consistent with the hypothesis that the composition of the dietary fiber determines its adsorptive properties and that there may be important differences between the major dietary fibers of South Pacific and European food plants.


Potentials of biodegraded cashew pomace for cake baking. The use of biodegraded cashew pomace processed into flour for cake baking was investigated. The physico-chemical changes during the submerged fermentation of the pomace and the organoleptic qualities of the composite cake were also monitored. There was an increase of about 50% in protein content of the pomace after 96 h of fermentation. However, a reduction of about 61% in the total microbial count after 24 h was due to the toxic effect of the organic acids on the microbial cells during fermentation. The cashew flour had high crude fibre (ca. 20-33%) and carbohydrate (ca. 16-47%) values. The composite cake made from a 10:90 combination of 96 h-degraded cashew flour/wheat flour respectively was the most accepted. The cake which had a specific volume of 0.53 ml/g lost 11.1% moisture when 38 g of its batter was exposed to 190 degrees C for 10 minutes. This cake had a calorie value of 293.8/100 g and may be useful in feeding diabetic patients who require low carbohydrate foods.


Effect of cereal-legume chapati diets on absorption and retention of calcium. Wheat chapati, wheat + Bengal gram (80:20 and 70:30) chapatis and casein diets, at 10 percent protein level, were fed to rats for 12 weeks to study the effect on calcium utilization. The supplementation of Bengal gram to wheat diets significantly improved the calcium absorption. The urinary calcium excretion in wheat + Bengal gram chapati diets was significantly less than that of wheat chapati diet fed group. The in vivo 45Ca absorption (CPM/100 microliters serum) was also less in wheat chapati group as compared to wheat + Bengal gram chapati diets fed groups. The supplementation of legumes to cereals improved the calcium utilization and it may be concluded that there will be no risk of occurrence of protein induced hypercalciuria.


Studies on the effect of dry Sundakai (Solanum torvum) powder supplementation on lipid profile, glycated proteins and amino acids in non-insulin dependent diabetic patients. The effect of dry Sundakai powder supplementation (7 g providing 1.23 g of crude fibre) on glycemic control, lipidemic control, total amino acids and uronic acid was studied on 30 non-insulin dependent diabetes mellitus patients. All the patients were on hypoglycemic drugs. The above parameters were monitored at day 1, 15 and 30 days. After one month of fibre supplementation, no significant changes were observed with respect to glucose, lipid profile, glycated proteins, total amino acids and uronic acid levels in these subjects.