Treatment with a non-toxic, self-replicating anti-prion delays or prevents prion disease in vivo.

Transmissible spongiform encephalopathies (TSEs) are fatal neurological disorders caused by prions, which are composed of a misfolded protein (PrPSc) that self-propagates in the brain of infected individuals by converting the normal prion protein (PrPC) into the pathological isoform. Here, we report a novel experimental strategy for preventing prion disease based on producing a self-replicating, but innocuous PrPSc-like form, termed anti-prion, which can compete with the replication of pathogenic prions. Our results show that a prophylactic inoculation of prion-infected animals with an anti-prion delays the onset of the disease and in some animals completely prevents the development of clinical symptoms and brain damage. The data indicate that a single injection of the anti-prion eliminated ~99% of the infectivity associated to pathogenic prions. Furthermore, this treatment caused significant changes in the profile of regional PrPSc deposition in the brains of animals that were treated, but still succumbed to the disease. Our findings provide new insights for a mechanistic understanding of prion replication and support the concept that prion replication can be separated from toxicity, providing a novel target for therapeutic intervention.

Molecular interaction between type 2 diabetes and Alzheimer's disease through cross-seeding of protein misfolding.

Numerous epidemiological studies have shown a significantly higher risk for development of Alzheimer's disease (AD) in patients affected by type 2 diabetes (T2D), but the molecular mechanism responsible for this association is presently unknown. Both diseases are considered protein misfolding disorders associated with the accumulation of protein aggregates; amyloid-beta (Aß) and tau in the brain during AD, and islet amyloid polypeptide (IAPP) in pancreatic islets in T2D. Formation and accumulation of these proteins follows a seeding-nucleation model, where a misfolded aggregate or 'seed' promotes the rapid misfolding and aggregation of the native protein. Our underlying hypothesis is that misfolded IAPP produced in T2D potentiates AD pathology by cross-seeding Aß, providing a molecular explanation for the link between these diseases. Here, we examined how misfolded IAPP affects Aß aggregation and AD pathology in vitro and in vivo. We observed that addition of IAPP seeds accelerates Aß aggregation in vitro in a seeding-like manner and the resulting fibrils are composed of both peptides. Transgenic animals expressing both human proteins exhibited exacerbated AD-like pathology compared with AD transgenic mice or AD transgenic animals with type 1 diabetes (T1D). Remarkably, IAPP colocalized with amyloid plaques in brain parenchymal deposits, suggesting that these peptides may directly interact and aggravate the disease. Furthermore, inoculation of pancreatic IAPP aggregates into the brains of AD transgenic mice resulted in more severe AD pathology and significantly greater memory impairments than untreated animals. These data provide a proof-of-concept for a new disease mechanism involving the interaction of misfolded proteins through cross-seeding events which may contribute to accelerate or exacerbate disease pathogenesis. Our findings could shed light on understanding the linkage between T2D and AD, two of the most prevalent protein misfolding disorders.

Decolorization and biogas production by an anaerobic consortium: effect of different azo dyes and quinoid redox mediators.

The inhibitory effect of azo dyes and quinoid compounds on an anaerobic consortium was evaluated during a decolorization process and biogas production. In addition, the impact of quinoid compounds such as lawsone (LAW) and anthraquinone-2,6-disulfonate (AQDS) on the rate of decolorization of Direct Blue 71 (DB71) was assessed. The anaerobic consortium was not completely inhibited under all tested dye concentrations (0.1-2 mmol l(-1)), evidenced by an active decolorization process and biogas production. The presence of quinoid compounds at different concentrations (4, 8, and 12 mmol l(-1)) also inhibited biogas production compared to the control incubated without the quinoid compounds. In summary, the anaerobic consortium was affected to a greater extent by increasing the quantity of azo dyes or quinoid compounds. Nevertheless, at a lower concentration (1 mmol l(-1)) of quinoid compounds, the anaerobic consortium effectively decolorized 2 mmol l(-1) of DB71, increasing up to 5.2- and 20.4-fold the rate of decolorization with AQDS and LAW, respectively, compared to the control lacking quinoid compounds.

First Report of Powdery Mildew (Pseudoidium anacardii) of Mango Trees in Sinaloa, Mexico.

Powdery mildew of mango is an important disease in Mexico's northern Sinaloa state. Identification of the causal fungal agent has been hindered by the absence of information regarding its teleomorph, as well as a detailed morphometric analysis of the anamorph and molecular characterization. The first symptoms of the disease appear in mango inflorescences of early February, and it subsequently affects young fruits. The disease progresses during March and early April, causing significant fruit abortion and a scabby appearance in a high percentage of fruits that remain attached to the trees. We observed the disease on inflorescences but not in leaves during our sampling period. Powdery mildew specimens were collected during 2011 and 2012 and included Kent and Keith varieties from commercial orchards, and creole materials from backyards of private residences in the Ahome and Fuerte Counties of northern Sinaloa, Mexico. Symptomatic inflorescences were analyzed morphologically. Conidiophores and conidia were prepared by touching the whitish lesions with clear adhesive tape, which was then placed over microscope slides with a drop of distilled water and observed under a compound microscope. The anamorph structures of the pathogen were measured. The mycelium was septate and ramified on the surface of the host, forming a dense coat of branching hyphae. The mycelium had a diameter of 2.5 to 8.7 µm; conidiophores (Pseudoidium type) emerged from the superficial mycelium, were unbranched, and consisted of 1 to 3 cells with conidia forming singly from the apex. The length of the conidiophores varied from 30.0 to 77.5 µm; the foot cell of the conidiophores was straight, 10.0 to 47.5 µm long and with a diameter of 5.0 to 15.5 µm across its midpoint. Conidia without fibrosin bodies were borne singly, and were ellipsoid/ovoid, 22.5 to 46.2 µm long and 15.0 to 27.5 µm wide. Eighty percent of the germ tubes were forked (lobed); the rest were simple, emerged from the end, and were occasionally on the side of the conidia. Germ tubes ranged from 2.0 to 7.2 µm at the midpoint. The surface of the conidia appeared smooth under the scanning electron microscope, and elliptical conidia appeared constricted at their ends; this, however, was not observed in the ovoid conidia. In both cases, the terminal end of the conidia was smooth. The teleomorph was not found. Molecular and phylogenetic analysis of the ITS rDNA (2) region showed that samples are closely related to specimens of Pseudoidium anacardii (1) (teleomorph: Erysiphe quercicola [4]) collected from mango trees in diverse countries. Measurements of somatic and asexual structures are in agreement with descriptions of P. anachardii (formerly known as Oidium mangiferae) from India (3). The nucleotide sequences derived from this research were deposited in GenBank (Accession Nos. JX893951 to JX893957). To our knowledge, this is the first report of P. anacardii associated to mango inflorescences in Sinaloa, Mexico. Due to the economic importance of powdery mildew of mango trees in Sinaloa, future research directions should focus on finding the teleomorph of the fungus to support its identity. References: (1) U. Braun and R. T. A. Cook CBS Biodiversity Series No. 11, 2012. (2) S. Limkaisang et al. Mycoscience 47:327, 2006. (3) O. Prakash and K. C. Srivastava. Mango diseases and their management. A World Review Today and Tomorrow Publishers. New Delhi, India, 1987. (4) S. Takamatsu et al. Mycol. Res. 111:809, 2007.

Sex-specifics of ECT outcome.

OBJECTIVE: Electroconvulsive therapy (ECT) is the most effective treatment for patients with severe major depressive disorder (MDD). Given the known sex differences in MDD, improved knowledge may provide more sex-specific recommendations in clinical guidelines and improve outcome. In the present study we examine sex differences in ECT outcome and its predictors. METHODS: Clinical data from 20 independent sites participating in the Global ECT-MRI Research Collaboration (GEMRIC) were obtained for analysis, totaling 500 patients with MDD (58.6 % women) with a mean age of 54.8 years. Severity of depression before and after ECT was assessed with validated depression scales. Remission was defined as a HAM-D score of 7 points or below after ECT. Variables associated with remission were selected based on literature (i.e. depression severity at baseline, age, duration of index episode, and presence of psychotic symptoms). RESULTS: Remission rates of ECT were independent of sex, 48.0 % in women and 45.7 % in men (X2(1) = 0.2, p = 0.70). In the logistic regression analyses, a shorter index duration was identified as a sex-specific predictor for ECT outcome in women (X2(1) = 7.05, p = 0.01). The corresponding predictive margins did show overlapping confidence intervals for men and women. CONCLUSION: The evidence provided by our study suggests that ECT as a biological treatment for MDD is equally effective in women and men. A shorter duration of index episode was an additional sex- specific predictor for remission in women. Future research should establish whether the confidence intervals for the corresponding predictive margins are overlapping, as we find, or not.

Lesion environments direct transplanted neural progenitors towards a wound repair astroglial phenotype in mice.

Neural progenitor cells (NPC) represent potential cell transplantation therapies for CNS injuries. To understand how lesion environments influence transplanted NPC fate in vivo, we derived NPC expressing a ribosomal protein-hemagglutinin tag (RiboTag) for transcriptional profiling of transplanted NPC. Here, we show that NPC grafted into uninjured mouse CNS generate cells that are transcriptionally similar to healthy astrocytes and oligodendrocyte lineages. In striking contrast, NPC transplanted into subacute CNS lesions after stroke or spinal cord injury in mice generate cells that share transcriptional, morphological and functional features with newly proliferated host astroglia that restrict inflammation and fibrosis and isolate lesions from adjacent viable neural tissue. Our findings reveal overlapping differentiation potentials of grafted NPC and proliferating host astrocytes; and show that in the absence of other interventions, non-cell autonomous cues in subacute CNS lesions direct the differentiation of grafted NPC towards a naturally occurring wound repair astroglial phenotype.

The quinohaemoprotein alcohol dehydrogenase from Gluconacetobacter xylinus: molecular and catalytic properties.

Gluconacetobacter xylinus possesses a constitutive membrane-bound oxidase system for the use of ethanol. Its alcohol dehydrogenase complex (ADH) was purified to homogeneity and characterized. It is a 119-kDa heterodimer (68 and 41 kDa subunits). The peroxidase reaction confirmed the presence of haem C in both subunits. Four cytochromes c per enzyme were determined by pyridine hemochrome spectroscopy. Redox titrations of the purified ADH revealed the presence of four haem c redox centers, with apparent mid-point potential values (Em(7)) of -33, +55, +132 and +310 mV, respectively. The ADH complex contains one mol of pyrroloquinoline quinone as determined by HPLC. The enzyme was purified in full reduced state; oxidation was induced by potassium ferricyanide and substrate restores full reduction. Activity responses to pH were sharp, showing two distinct optimal pH values (i.e. pH 5.5 and 6.5) depending on the electron acceptor used. Purified ADH oxidizes primary alcohols (C2-C6) but not methanol. Noteworthy, aliphatic aldehydes (C1-C4) were also good substrates. Myxothiazol and antymicin A were powerful inhibitors of the purified ADH complex, most likely acting at the ubiquinone acceptor site in subunit II.

Concentraciones de oxígeno intermitentes tienen impacto en el desarrollo postnatal temprano de ratas Sprague-Dawley: efectos en CA1 del hipocampo / Intermittent oxygen concentrations have an impact on the early postnatal development of Sprague-Dawley rats: effects on hippocampal CA1

El periodo postnatal temprano se caracteriza por rápido crecimiento cerebral, posiblemente relacionado con variaciones del oxígeno tisular. Esto ha motivado el estudio de protocolos que suministran diferentes concentraciones de oxígeno intermitentes, para observar sus efectos morfológicos y cerebrales. Se utilizaron 52 crías de ratas Sprague Dawley, distribuidas en igual número a cuatro grupos experimentales, Control (C, 21 %O2), Hipoxia Intermitente (HI, 11 %O2), Hiperoxia Intermitente (HOI, 30 %O2) e Hipoxia Hiperoxia Intermitente (HHI, 11 % -30 %O2). Los protocolos consideraron 5 ciclos de 5 minutos de dosificación, durante 50 minutos diarios. Se realizó en una cámara semihermética entre los días 5 al 11 postnatales. Las evaluaciones de crecimiento corporal y cuantificación neuronal, se realizaron en las crías macho, en el día 28 postnatal. El peso corporal en el grupo hipoxia intermitente mostró diferencias significativas respecto al grupo hiperoxia intermitente (HI vs HOI, p<0,01) y al grupo hipoxia-hiperoxia Intermitente (HI vs HHI, p< 0,001). La talla corporal disminuyó en el grupo hipoxia-hiperoxia intermitente con diferencias significativas respecto del grupo control (C vs HHI, p<0,05) y respecto del grupo hipoxia intermitente (HHI vs HI, p< 0,01). El conteo neuronal en el área CA1 del hipocampo aumentó en el grupo hipoxia intermitente con diferencias significativas respecto a los grupos control (C vs HI; p<0,05), al grupo hiperoxia intermitente (HI vs HOI; p<0,001) y al grupo hipoxia-hiperoxia intermitente (HI vs HHI; p<0,001). Finalmente, el grupo hipoxia- hiperoxia Intermitente disminuyó significativamente en la cantidad de neuronas en comparación al grupo hiperoxia intermitente (HHI vs HOI; p<0,001). La hipoxia intermitente mostró resultados beneficiosos en el crecimiento corporal y cantidad de neuronas en el área CA1 del hipocampo, en contraste, la hipoxia hiperoxia intermitente experimentó resultados adversos con disminución de estas variables, en el periodo postnatal temprano de la rata.

SUMMARY: The early postnatal period is characterized by rapid brain growth, possibly related to variations in tissue oxygen. This has motivated the study of protocols that supply different intermittent oxygen concentrations, to observe their morphological and cerebral effects. Fifty-two pups Sprague-Dawley rats were distributed in equal numbers into four experimental groups, Control (C, 21 %O), Intermittent Hypoxia (HI, 11 %O), Intermittent Hyperoxia (HOI, 30 %O2) and Intermittent Hypoxia Hyperoxia (HHI, 11 % - 30 %O2). The protocols considered 5 cycles of 5 min of dosing, for 50 min diary. It was performed in a semi- hermetic chamber between 5 to 11postnatal days. The evaluations of body growth and neuronal quantification were analyzed in male pups, on postnatal day 28. Body weight in the intermittent hypoxia group showed significant differences compared to the intermittent hyperoxia group (HI vs HOI, p<0.01) and the intermittent hypoxia- hyperoxia group (HI vs HHI, p<0.001). Body size decreased in the Intermittent hypoxia-hyperoxia group with significant differences compared to the control group (C vs HHI, p<0.05) and with respect to the intermittent hypoxia group (HHI vs HI, p<0.01). The neuronal count in the area CA1 of the hippocampus increased in the intermittent hypoxia group with significant differences compared to the control groups (C vs HI; p<0.05), to the intermittent hyperoxia group (HI vs HOI; p< 0.001) and the intermittent hypoxia-hyperoxia group (HI vs HHI; p<0.001). Finally, the intermittent hypoxia- hyperoxia group decreased significantly in the number of neurons compared with the intermittent hyperoxia group (HHI vs HOI; p<0.001). Intermittent hypoxia showed beneficial results in body growth and the number of neurons in the CA1 area of the hippocampus, in contrast, intermittent hypoxia-hyperoxia experienced adverse results with a decrease in these variables, in the early postnatal period of the rat.

sn-1,2-diacylglycerol and choline increase after fertilization in Xenopus laevis.

sn-1,2-Diacylglycerol (DAG) mass and translocation of protein kinase C alpha and beta to a membrane fraction increased approximately 7 min after insemination of Xenopus laevis eggs. The DAG mass increase of 48 pmol (from 62 to 110 pmol/cell) was greater than that for inositol 1,4,5-trisphosphate (IP3; an increase of approximately 170 fmol or approximately 280-fold smaller than the DAG increase), and DAG peaks approximately 5 min after IP3. Choline mass (a measure of phosphatidyl choline-specific phospholipase D) also peaked before DAG and the choline increase (134 pmol/cell) was greater than that of DAG. There was no detectable change in phosphocholine mass (a measure of phosphatidylcholine-specific phospholipase C). During first cleavage, DAG decreased, PKC translocation was low, and choline increased and peaked (whereas published work shows an increase in IP3 mass). Artificial elevation of intracellular calcium ([Ca2+]i) increased DAG levels but prevention of the [Ca2+]i increase after fertilization did not block DAG production. Thus, sperm stimulate production of DAG and choline through [Ca2+]i-independent and [Ca2+]i-dependent paths.

Unilateral retinitis pigmentosa. A case report. / Retinitis pigmentosa unilateral a propósito de un caso.

CLINICAL CASE: A 27-year-old woman with a history of nyctalopia and constriction of visual field of the right eye. The ophthalmological examination showed a visual field and electroretinogram that were compatible with unilateral retinitis pigmentosa (RP). After a one year follow-up, the unilateral condition remained. DISCUSSION: Unilateral retinitis pigmentosa is a rare condition, with a frequency between 0.2%-5% of the RP. It mainly affects women and older age groups than bilateral RP. For a definitive diagnosis, it is necessary to have a funduscopy and electroretinogram (ERG) altered unilaterally, and exclude infectious, inflammatory, and vascular causes.

COVID prolongado / Long COVID

Existen múltiples reportes de manifestaciones persistentes en pacientes que cursaron infecciones por SARS-CoV-2, independiente de su gravedad, configurando el síndrome de COVID-19 prolongado. No existe una definición consensuada de este síndrome, cuya patogenia pareciera ser multifactorial. Considerando las más de 500 millones de infecciones en todo el mundo, este síndrome pudiese incidir en una insospechada y prolongada carga sobre los sistemas sanitarios. Reportes recientes han asociado a la vacunación con esquema primario completo como una asociación protectora para el desarrollo de COVID-19 prolongado, transformándose en otro beneficio poblacional asociado a las vacunas.(AU)

There are multiple reports of persistent manifestations in patients who had SARS-CoV-2 infections, regardless of their severity, configuring the prolonged COVID-19 syndrome. There is no agreed definition of this syndrome whose pathogenesis seems to be multifactorial. Considering the more than 500 million infections worldwide, this syndrome could have an unsuspected and prolonged burden on health systems . Recent reports have associated vaccination with a complete primary schedule as a protective association with the development of prolonged COVID-19, becoming another population benefit associated with vaccines.(AU)

Aerial pesticide application causes DNA damage in pilots from Sinaloa, Mexico.

The use of pesticides in agricultural production originates residues in the environment where they are applied. Pesticide aerial application is a frequent source of exposure to pesticides by persons dedicated to agricultural practices and those living in neighboring communities of sprayed fields. The aim of the study was to assess the genotoxic effects of pesticides in workers occupationally exposed to these chemicals during their aerial application to agricultural fields of Sinaloa, Mexico. The study involved 30 pilots of airplanes used to apply pesticides via aerial application and 30 unexposed controls. Damage was evaluated through the micronucleus assay and by other nuclear abnormalities in epithelial cells of oral mucosa. The highest frequency ratios (FR) equal to 269.5 corresponded to binucleated cells followed by 54.2, corresponding to cells with pyknotic nuclei, 45.2 of cells with chromatin condensation, 3.7 of cells with broken-egg, 3.6 of cells with micronucleus, and 2.0 of karyolytic cells. Age, worked time, smoking, and alcohol consumption did not have significant influence on nuclear abnormalities in the pilots studied. Pesticide exposure was the main factor for nuclear abnormality results and DNA damage. Marked genotoxic damage was developed even in younger pilots with 2 years of short working period, caused by their daily occupational exposure to pesticides.

Effect of obese living donors on the outcome and metabolic features in recipients of kidney transplantation.

Living kidney donors must be evaluated carefully focusing on risk factors for long-term complications. Our transplant center performs 70% of its kidney transplantations from living sources including 19.9% obese donors. We evaluated the long-term follow-up of recipients of organs from 37 living donor patients with obesity defined by a body mass index (BMI) > 30 kg/m2. We compared this group with a control group of normal BMI before donation. The follow-up was 50.8 +/- 28.5 months. We observed a lower glomerular filtration rate among organs from obese versus non-obese donors. At the same time we reviewed percentage of acute rejection episodes (ARE), primary allograft function, and surgical complications we observed incidence of ARE higher among the group who received kidneys from obese donors.

Prevalence of erectile dysfunction in kidney transplant recipients.

Prevalence and severity of erectile dysfunction increase with advancing age. Patients with end-stage renal disease (ESRD) experience disturbances in erectile function related to organic factors including as uremia, hypertension, endocrine, and nonorganic factors like depression. Recipients of kidney transplants show a high prevalence of erectile dysfunction, 32.2% to 50.7%. We conducted a study of the prevalence of erectile dysfunction among male renal transplant recipients using the International Index of Erectile Function. Among 182 men with kidney transplantations, there were 89 recipients (48.9%) with erectile dysfunction; 60 recipients had normal sexual function (32.9%); and whereas 33 recipients had no sexual activity.

Variante Ómicron SARS-CoV-2: una nueva variante de preocupación / Ómicron SARS-CoV-2 variant: anew variant of concern

Ha surgido una nueva variante de preocupación de SARS-CoV-2, cuyos efectos en la evolución de la pandemia parecen inciertos. Sin embargo, ha comenzado a surgir evidencia con respecto al comportamiento viral en cuanto a su transmisibilidad, unión a receptor de la célula hospedadora y escape del sistema inmune. Presentamos una revisión actualizada de los datos existentes en la literatura respecto a los aspectos microbiológicos y epidemiológicos que pueden ayudarnos a comprender las futuras investigaciones en esta variante.(AU)

A new variant of concern for SARS-CoV-2 has emerged, the effects of which on the evolution of the pandemic appear uncertain. However, evidence has begun to emerge regarding viral behavior in terms of its transmissibility, receptor binding on the host cell, and escape from the immune system. We present an updated review of the existing data in the literature regarding the microbiological and epidemiological aspects that can help us understand future research on this variant.(AU)

Electroconvulsive therapy and structural neuroplasticity in neocortical, limbic and paralimbic cortex.

Electroconvulsive therapy (ECT) is a highly effective and rapidly acting treatment for severe depression. To understand the biological bases of therapeutic response, we examined variations in cortical thickness from magnetic resonance imaging (MRI) data in 29 patients scanned at three time points during an ECT treatment index series and in 29 controls at two time points. Changes in thickness across time and with symptom improvement were evaluated at high spatial resolution across the cortex and within discrete cortical regions of interest. Patients showed increased thickness over the course of ECT in the bilateral anterior cingulate cortex (ACC), inferior and superior temporal, parahippocampal, entorhinal and fusiform cortex and in distributed prefrontal areas. No changes across time occurred in controls. In temporal and fusiform regions showing significant ECT effects, thickness differed between patients and controls at baseline and change in thickness related to therapeutic response in patients. In the ACC, these relationships occurred in treatment responders only, and thickness measured soon after treatment initiation predicted the overall ECT response. ECT leads to widespread neuroplasticity in neocortical, limbic and paralimbic regions and changes relate to the extent of antidepressant response. Variations in ACC thickness, which discriminate treatment responders and predict response early in the course of ECT, may represent a biomarker of overall clinical outcome. Because post-mortem studies show focal reductions in glial density and neuronal size in patients with severe depression, ECT-related increases in thickness may be attributable to neuroplastic processes affecting the size and/or density of neurons and glia and their connections.

Correlation of Allograft Weight to Recipient Body Weight Index on Renal Function in Kidney Transplantation.

BACKGROUND: The function of renal transplant grafts can be modified by many factors. In one study of graft weight/weight of the recipient, it was concluded to avoid renal transplantation in patients with kidneys with a low ratio between the graft and recipient weight (<2.5 g/kg). OBJECTIVE: The objective of this study was to describe the association between renal allograft weight and renal function 1 month after renal transplantation in the Mexican population. MATERIAL AND METHODS: We studied patients who underwent transplantation from living or cadaveric donors with 1 month of follow-up with a functioning graft. An observational, retrospective, analytic study from January 1, 2014 to November 1, 2014 was conducted. Graft weight, donor weight, recipient weight, age, donor gender, recipient creatinine, and renal function were recorded. RESULTS: A total of 88 patients were included: 35 women (39.8%) and 53 men (60.2%). Sixty (68%) received kidneys from living donors and 28 (31.8%) from deceased donors. Mean recipient body mass index (BMI) was 24.7 (±2.6). Mean graft weight was 152 g (±33.9). Creatinine at 1 month post-transplantation was 1.6 mg/dL (±2.0). Using a linear regression model cold ischemia time was related to serum creatinine at 1 month post-transplantation (P = .020). Using multivariate analysis, significance was observed with respect to these indexes and recipient renal function. Recipient gender also was related and showed statistical significance (P = .015). CONCLUSIONS: Renal graft function depends on many factors including the amount of functional renal mass and nephrons required according to the recipient's weight. The donor kidney weigt (DKW) / receptor body weigt (RBW) index should be considered as selection criteria of donors.

The antidiabetic drug metformin elevates receptor tyrosine kinase activity and inositol 1,4,5-trisphosphate mass in Xenopus oocytes.

Although metformin is an important antidiabetic, its mechanism of action is still unknown. To study its mechanism, we examined metformin stimulation of insulin action on the Xenopus oocyte. Similar to therapeutic concentrations, maximal stimulation of insulin-induced meiotic cell division was achieved at about 1-10 microg/ml (or 7.7-77 /microM) metformin. An equivalent concentration of metformin was required to elevate receptor tyrosine kinase activity (in whole cells or a membrane-cortex preparation) and, through this tyrosine kinase activation, inositol 1,4,5-trisphosphate (IP3) production. With whole cells, the preincubation time for metformin stimulation of insulin action (approximately 1 h) was equivalent to the time required for metformin to maximize tyrosine phosphorylation and raise IP3, levels. With the membrane-cortex preparation, metformin was active within minutes; thus, metformin may act at an intracellular site. Since metformin can increase IP3, mass, we prevented elevation of calcium by prior microinjection of a calcium chelator or heparin (a drug that inhibits IP3 binding to the IP3 receptor). Both the chelator and heparin blocked metformin stimulation of insulin action on whole cells. Since microinjection of IP3, also stimulates insulin action, metformin may stimulate insulin action by elevation of intracellular calcium in addition to activation of the receptor tyrosine kinase.

Quantitation of soluble HLA class I heterodimers and beta 2-microglobulin in patients with active pulmonary tuberculosis.

Beta 2m serum levels have been shown to be increased in patients with tuberculosis and HIV infection. We determined the stability of beta 2m and of sHLA-I dimers in serum, and then determined the levels of both molecules in 60 non-HIV-infected patients with active pulmonary tuberculosis and in 55 adult controls. The levels of sHLA-I in samples kept at room temperature declined by 8% at 30 minutes, 16% at 60 minutes, and 36% at 120 minutes. Beta 2m levels remained stable at all times tested. Mean sHLA-I levels were 0.99 +/- 0.16 micrograms/ml in controls and 1.34 +/- 0.11 micrograms/ml in patients with tuberculosis (P < 0.0001). Beta 2m levels were 1.23 +/- 0.26 micrograms/ml in controls and 2.26 +/- 0.64 micrograms/ml in patients with tuberculosis (P < 0.0001). All patients with tuberculosis had elevation of sHLA-I and/or beta 2m above 1 standard deviation of normal values. However, there was no correlation between sHLA-I and beta 2m levels in individual samples. Evaluation of sHLA-I holds the promise of further understanding of the biology and genetic regulation of the immune response to mycobacterial infection.