mTOR-dependent loss of PON1 secretion and antiphospholipid autoantibody production underlie autoimmunity-mediated cirrhosis in transaldolase deficiency.

Transaldolase deficiency predisposes to chronic liver disease progressing from cirrhosis to hepatocellular carcinoma (HCC). Transition from cirrhosis to hepatocarcinogenesis depends on mitochondrial oxidative stress, as controlled by cytosolic aldose metabolism through the pentose phosphate pathway (PPP). Progression to HCC is critically dependent on NADPH depletion and polyol buildup by aldose reductase (AR), while this enzyme protects from carbon trapping in the PPP and growth restriction in TAL deficiency. Although AR inactivation blocked susceptibility to hepatocarcinogenesis, it enhanced growth restriction, carbon trapping in the non-oxidative branch of the PPP and failed to reverse the depletion of glucose 6-phosphate (G6P) and liver cirrhosis. Here, we show that inactivation of the TAL-AR axis results in metabolic stress characterized by reduced mitophagy, enhanced overall autophagy, activation of the mechanistic target of rapamycin (mTOR), diminished glycosylation and secretion of paraoxonase 1 (PON1), production of antiphospholipid autoantibodies (aPL), loss of CD161+ NK cells, and expansion of CD38+ Ito cells, which are responsive to treatment with rapamycin in vivo. The present study thus identifies glycosylation and secretion of PON1 and aPL production as mTOR-dependent regulatory checkpoints of autoimmunity underlying liver cirrhosis in TAL deficiency.

Predicting onset of disease progression using temporal disease occurrence networks.

OBJECTIVE: Early recognition and prevention are crucial for reducing the risk of disease progression. This study aimed to develop a novel technique based on a temporal disease occurrence network to analyze and predict disease progression. METHODS: This study used a total of 3.9 million patient records. Patient health records were transformed into temporal disease occurrence networks, and a supervised depth first search was used to find frequent disease sequences to predict the onset of disease progression. The diseases represented nodes in the network and paths between nodes represented edges that co-occurred in a patient cohort with temporal order. The node and edge level attributes contained meta-information about patients' gender, age group, and identity as labels where the disease occurred. The node and edge level attributes guided the depth first search to identify frequent disease occurrences in specific genders and age groups. The patient history was used to match the most frequent disease occurrences and then the obtained sequences were merged together to generate a ranked list of diseases with their conditional probability and relative risk. RESULTS: The study found that the proposed method had improved performance compared to other methods. Specifically, when predicting a single disease, the method achieved an area under the receiver operating characteristic curve (AUC) of 0.65 and an F1-score of 0.11. When predicting a set of diseases relative to ground truth, the method achieved an AUC of 0.68 and an F1-score of 0.13. CONCLUSION: The ranked list generated by the proposed method, which includes the probability of occurrence and relative risk score, can provide physicians with valuable information about the sequential development of diseases in patients. This information can help physicians to take preventive measures in a timely manner, based on the best available information.

Cytosolic aldose metabolism contributes to progression from cirrhosis to hepatocarcinogenesis.

Oxidative stress modulates carcinogenesis in the liver; however, direct evidence for metabolic control of oxidative stress during pathogenesis, particularly, of progression from cirrhosis to hepatocellular carcinoma (HCC), has been lacking. Deficiency of transaldolase (TAL), a rate-limiting enzyme of the non-oxidative branch of the pentose phosphate pathway (PPP), restricts growth and predisposes to cirrhosis and HCC in mice and humans. Here, we show that mitochondrial oxidative stress and progression from cirrhosis to HCC and acetaminophen-induced liver necrosis are critically dependent on NADPH depletion and polyol buildup by aldose reductase (AR), while this enzyme protects from carbon trapping in the PPP and growth restriction in TAL deficiency. Both TAL and AR are confined to the cytosol; however, their inactivation distorts mitochondrial redox homeostasis in opposite directions. The results suggest that AR acts as a rheostat of carbon recycling and NADPH output of the PPP with broad implications for disease progression from cirrhosis to HCC.

Utility of sonographic transcerebellar diameter in the assessment of gestational age in normal and intrauterine growth-retarded fetuses.

BACKGROUND: The fetal cerebellum has been shown to be least affected by external pressures and molding during pregnancy and therefore might provide more accurate estimation of GA. AIMS: To study the utility of transcerebellar diameter (TCD) measured by ultrasound for the detection of GA in normal and intrauterine growth-retarded (IUGR) fetuses. SUBJECTS AND METHODS: This cross-sectional study comprised 500 antenatal patients with a GA between 14 and 39 weeks and who were certain of their last menstrual periods. The TCD was measured ultrasonographically and the corresponding GA was determined. The GA was also determined with other customarily used sonographic parameters such as biparietal diameter (BPD), head circumference (HC), abdominal circumference (AC), and femur length (FL) and compared with TCD. Data of normal pregnancy patients was used to formulate nomograms by taking the 5th, 50th, and 95th percentile measurements. TCD to AC ratio was also calculated in both normal (n = 424) and IUGR pregnancies (n = 76). RESULTS: TCD showed significant correlation with gestational age (GA) measured by last menstrual period (LMP) as well as with GA calculated with other biometric fetal parameters. TCD also showed significant correlation with GA in normal (R2 = 0.979) as well as with IUGR pregnancies (R2 = 0.942). TCD to AC ratio remained fairly constant in normal pregnancies while it was increased in IUGR pregnancies. CONCLUSIONS: TCD and TCD/AC ratio can be employed as an objective parameter to establish the GA in normal as well as IUGR pregnancy cases.

Skull Base Osteomyelitis: A Comprehensive Imaging Review.

Skull base osteomyelitis is a relatively rare condition, generally occurring as a complication of advanced otologic or sinus infection in immunocompromised patients. Skull base osteomyelitis is generally divided into 2 broad categories: typical and atypical. Typical skull base osteomyelitis occurs secondary to uncontrolled infection of the temporal bone region, most often from necrotizing external otitis caused by Pseudomonas aeruginosa in a patient with diabetes. Atypical skull base osteomyelitis occurs in the absence of obvious temporal bone infection or external auditory canal infection. It may be secondary to advanced sinusitis or deep face infection or might occur in the absence of a known local source of infection. Atypical skull base osteomyelitis preferentially affects the central skull base and can be caused by bacterial or fungal infections. Clinically, typical skull base osteomyelitis presents with signs and symptoms of otitis externa or other temporal bone infection. Both typical and atypical forms can produce nonspecific symptoms including headache and fever, and progress to cranial neuropathies and meningitis. Early diagnosis can be difficult both clinically and radiologically, and the diagnosis is often delayed. Radiologic evaluation plays a critical role in the diagnosis of skull base osteomyelitis, with CT and MR imaging serving complementary roles. CT best demonstrates cortical and trabecular destruction of bone. MR imaging is best for determining the overall extent of disease and best demonstrates involvement of marrow space and extraosseous soft tissue. Nuclear medicine studies can also be contributory to diagnosis and follow-up. The goal of this article was to review the basic pathophysiology, clinical findings, and key radiologic features of skull base osteomyelitis.

Upregulation of TET activity with ascorbic acid induces epigenetic modulation of lymphoma cells.

The Ten Eleven Translocation (TET) enzymes have been found to be mutated in both diffuse large B-cell (DLBCL) and peripheral T-cell (PTCL) lymphomas resulting in DNA hypermethylation. Recent studies in embryonal stem cells showed that ascorbic acid (AA) is a cofactor for TET with a binding site at the catalytic domain, and enhances TET activity. We hypothesized that AA could potentially enhance TET activity in lymphoma cells to cause DNA demethylation, reactivate expression of tumor suppressor genes and enhance chemosensitivity. We demonstrate in vitro that AA treatment of DLBCL and PTCL cells using AA concentrations achievable intravenously increased TET activity leading to DNA demethylation. This epigenetic effect is independent of hydrogen peroxide. AA treatment increased the expression of SMAD1, a tumor suppressor gene known to be suppressed by methylation, and increased chemosensitivity of lymphoma cells. Twenty-nine percent (10/34) of unselected lymphoma patients had plasma AA levels that were deficient suggesting an additional clinical mechanism of TET hypofunction. These data indicate that AA has the potential to modify TET function in lymphoma and enhance chemosensitivity. In addition, the AA deficiency seen in some patients may further impair TET function and contribute to resistance. Clinical trials testing intravenous AA with chemotherapy are warranted.

MCL-1 and BCL-xL-dependent resistance to the BCL-2 inhibitor ABT-199 can be overcome by preventing PI3K/AKT/mTOR activation in lymphoid malignancies.

Overexpression of anti-apoptotic BCL-2 family members is a hallmark of many lymphoid malignancies, including chronic lymphocytic leukemia (CLL) and non-Hodgkin lymphoma (NHL) that can be targeted with small molecule inhibitors. ABT-199 is a rationally designed BCL-2 homology (BH)-3 mimetic that specifically binds to BCL-2, but not to MCL-1 and BCL-xL. Although the thrombocytopenia that occurs with navitoclax treatment has not been a problem with ABT-199, clinical trials in CLL could benefit by lowering the ABT-199 concentration through targeting other survival pathways. In this study, we investigated the mechanisms of resistance that develops to ABT-199 therapy by generating ABT-199-resistant (ABT199-R) cell lines via chronic exposure of NHL cell lines to ABT-199. Acquired resistance resulted in substantial AKT activation and upregulation of MCL-1 and BCL-xL levels that sequestered BIM. ABT199-R cells exhibited increased MCL-1 stability and failed to activate BAX in response to ABT-199. The ABT-199 acquired and inherent resistant cells were sensitized to treatment with ABT-199 by inhibitors of the PI3K, AKT, and mTOR pathways, NVP-BEZ235 and GS-1101. NVP-BEZ235, a dual inhibitor of p-AKT and mTOR, reduced MCL-1 levels causing BIM release from MCL-1 and BCL-xL, thus leading to cell death by BAX activation. The PI3Kδ inhibitor GS-1101 (idelalisib) downregulated MCL-1 and sensitized ABT199-R cells through AKT-mediated BAX activation. A genetic approach, through siRNA-mediated down-regulation of AKT, MCL-1, and BCL-xL, significantly decreased cell survival, demonstrating the importance of these cell survival factors for ABT-199 resistance. Our findings suggest a novel mechanism that modulates the expression and activity of pro-survival proteins to confer treatment resistance that could be exploited by a rational combination therapeutic regimen that could be effective for treating lymphoid malignancies.

Multifocal skeletal tuberculosis involving the lumbar spine and iliac bone, mimicking a malignant bone tumour: a case report.

ABSTRACT: Multifocal osteoarticular tuberculosis is uncommon and accounts for approximately10% of all cases of osteoarticular tuberculosis. Concomitant vertebral and iliac bone tuberculosis has not been reported in the literature to date. We report one such case for its unusual clinical and radiological presentation, which had presentation more similar to a malignant bone tumour than an infection. KEY WORDS: Skeletal tuberculosis, multifocal osteoarticular tuberculosis.

Acidic stress promotes a glioma stem cell phenotype.

Malignant gliomas are lethal cancers that display cellular hierarchies with cancer stem cells at the apex. Glioma stem cells (GSCs) are not uniformly distributed, but rather located in specialized niches, suggesting that the cancer stem cell phenotype is regulated by the tumor microenvironment. Indeed, recent studies show that hypoxia and its molecular responses regulate cancer stem cell maintenance. We now demonstrate that acidic conditions, independent of restricted oxygen, promote the expression of GSC markers, self-renewal and tumor growth. GSCs exert paracrine effects on tumor growth through elaboration of angiogenic factors, and low pH conditions augment this expression associated with induction of hypoxia inducible factor 2α (HIF2α), a GSC-specific regulator. Induction of HIF2α and other GSC markers by acidic stress can be reverted by elevating pH in vitro, suggesting that raising intratumoral pH may be beneficial for targeting the GSC phenotype. Together, our results suggest that exposure to low pH promotes malignancy through the induction of a cancer stem cell phenotype, and that culturing cancer cells at lower pH reflective of endogenous tumor conditions may better retain the cellular heterogeneity found in tumors.

Biodeterioration in Emblica based Medicinal products and their Aflatoxin contamination.

In the present investigation, Emblica officinalis Gaerth. Syn Phyllanthus emblica linn. has been selected on account of its large scale use in manufacture of indigenous drugs. Among the various fungal, species, Aspergillus flavus was the most dominant and frequent species in most of food stuffs studied. The total population of Aspergillus. flavus was recorded in range of 20.30 × 10(3) - 89.50 × 10(3) colonies/g in dried Aonla, 8.00 - 42.25 × 10(3) colonies/g in dried Aonla, 8.00 - 42.25 × 10(3) colonies/g Triphala, 1.25 × 10(3) - 19.83 × 10(3) colonies/g in hair oil and 1.20- 27.00 × 10(3) colonies/g in Aonla syrup. Minimum 26.66% percentage incidence of aflatoxin contamination was recorded in samples where the frequency of Aspergillus flavus was 91.50%.

Primary spinal pleomorphic xanthoastrocytoma.

Pleomorphic xanthochromic astrocytoma primarily of the spinal cord is a rare entity. The case is possibly the fifth such report. Complete surgical excision is the essential requirement for good survival. In the absence of any clearly laid down protocols of adjuvant treatment, anecdotal reports support treatment with chemotherapy alone or both chemotherapy and radiotherapy.

Diuretic activity of the leaves of Coleus aromaticus Benth.

The aqueous and ethanolic extract of leaves of Coleus aromaticus was evaluated for diuretic activity. Both extracts were evaluated by determination of urine volume and electrolyte concentration in albino rats. Results revealed that both the aqueous and ethanolic extract at dose 500mgl kg showed significant diuretic activity by increasing the total volume of urine and concentration electrolyte. Furosamide (10 mg/kg) was used as reference drug while normal saline (0.9%) solution was used as control.

Functional characterization of a putative endoglucanase gene in the genome of Zymomonas mobilis.

The sequence of the putative endoglucanase gene ZMO1086 in the genome of Zymomonas mobilis showed a 40% similarity with known bacterial endoglucanase genes. The upstream region of this putative gene revealed the presence of characteristic promoter (-10 and -35 regions) and a Shine-Dalgarno region. The putative endoglucanase gene was poorly expressed from the native promoter of Z. mobilis and therefore the putative endoglucanase gene was cloned and expressed in Escherichia coli BL21. The overexpressed gene product CelA was purified to homogeneity and the optimal activity was observed at 30 degrees C and pH 6 respectively.

Dengue: A Clinicohaematological Profile.

BACKGROUND: Fifty cases of fever, clinically suspected to be dengue were studied. METHODS: Complete clinical, haematological evaluation and IgM capture assay was done. RESULT: 54% of patients clinically suspected to have dengue were positive for IgM antibodies by enzyme-linked immunosorbent assay (ELISA). The commonest clinical feature was fever with rash (85%). Thrombocytopenia was seen in 19% of patients only. One patient died of dengue shock syndrome (DSS). CONCLUSION: Out of the 27 cases of seropositive dengue there was one death due to dengue shock syndrome. Thrombocytopenia may not always be a feature of dengue.

In vitro antioxidant studies of the ethanolic extract of Tephrosia purpurea L.

The ethanolic extract of the root of Tephrosia purpurea was screened for in vitro antioxidant properties using standard procedures. The ethanolic extract exhibited IC(50) values of 132.31±8.79 and 405.22±15.09 respectively in DPPH and nitric acid radical inhibition assay. These values were slightly more than those obtained for ascorbic acid and rutin used as standard. The findings justify the therapeutic application of the plant in the indigenous system of medicine, augmenting its therapeutic value.

Antimicrobial Activity of Root Bark of Salacia reticulata.

Antimicrobial activity of chloroform and methanolic extracts of Salacia reticulata were tested against gram positive, gram negative and fungus strains using zone of inhibition and minimum inhibitory concentrations. It was observed that both extracts have inhibitory effect towards all microorganisms used in the test. Chloroform extract was more effective than methanolic extract.

Carvedilol titration in patients with congestive heart failure receiving inotropic therapy.

BACKGROUND: Carvedilol has been shown to improve morbidity and mortality in patients with congestive heart failure (CHF). There are limited data of carvedilol use in patients on inotrope therapy. We present our experience with carvedilol titration in New York Heart Association (NYHA) class IIIb/IV patients stabilized on milrinone therapy, as a nonrandomized study with a parallel control group of patients never on inotropes. These patients achieved volume control and stabilization of their symptoms during the course of milrinone therapy. METHODS AND RESULTS: Seventeen patients in class IIIb/IV CHF (group 1) on intermittent intravenous milrinone therapy and 15 patients in class II/IIIa compensated CHF (group 2) on standard triple heart failure therapy were titrated on carvedilol. Success and adverse events during titration were compared between the 2 groups. Fifteen (88%) patients in group 1 and 14 (93%) patients in group 2 were successfully titrated on carvedilol over 8.1 +/- 1.8 weeks and 6.7 +/- 2.8 weeks, respectively. The target dose of carvedilol (25 or 50 mg twice daily) was achieved in 13 (87%) patients (group 1) and 14 (93%) patients (group 2). Seven (47%) patients in group 1 and 4 (28%) patients in group 2 had adverse events during carvedilol titration. Eight (53%) patients in group 1 were weaned off milrinone over a period of 8.4 weeks after carvedilol titration, whereas the rest of the patients had reduction in the frequency of infusion. Ten (63%) patients in group 1 improved by one or more functional classes. CONCLUSIONS: Patients in NYHA class IIIb/IV who are treated with inotropic therapy can be titrated on carvedilol after reaching a stable state while on milrinone and standard oral drugs. Most of these patients can be successfully weaned off of milrinone or have decreased frequency of infusions and demonstrate improved functional status. Prospective randomized trials are required to evaluate these observations made in a limited number of patients in class IIIb and IV CHF because the combination of milrinone and beta-blockers has never been adequately evaluated.

Apoptosis of lung epithelial cells in response to TNF-alpha requires angiotensin II generation de novo.

Recent work from this laboratory demonstrated that apoptosis of pulmonary alveolar epithelial cells (AEC) in response to Fas requires angiotensin II (ANGII) generation de novo and binding to its receptor (Wang et al., 1999b, Am J Physiol Lung Cell Mol Physiol 277:L1245-L1250). These findings led us to hypothesize that a similar mechanism might be involved in the induction of AEC apoptosis by TNF-alpha. Apoptosis was detected by assessment of nuclear and chromatin morphology, increased activity of caspase 3, binding of annexin V, and by net cell loss inhibitable by the caspase inhibitor ZVAD-fmk. Purified human TNF-alpha induced dose-dependent apoptosis in primary type II pneumocytes isolated from rats or in the AEC-derived human lung carcinoma cell line A549. Apoptosis in response to TNF-alpha was inhibited in a dose-dependent manner by the nonselective ANGII receptor antagonist saralasin or by the nonthiol ACE inhibitor lisinopril; the inhibition of TNF-induced apoptosis was maximal at 50 microgram/ml saralasin (101% inhibition) and at 0.5 microgram/ml lisinopril (86% inhibition). In both cell culture models, purified TNF-alpha caused a significant increase in the mRNA for angiotensinogen (ANGEN), which was not expressed in unactivated cells. Transfection of primary cultures of rat AEC with antisense oligonucleotides against ANGEN mRNA inhibited the subsequent induction of TNF-stimulated apoptosis by 72% (P < 0.01). Exposure to TNF-alpha increased the concentration of ANGII in the serum-free extracellular medium by fivefold in A549 cell cultures and by 40-fold in primary AEC preparations; further, exposure to TNF-alpha for 40 h caused a net cell loss of 70%, which was completely abrogated by either the caspase inhibitor ZVAD-fmk, lisinopril, or saralasin. Apoptosis in response to TNF-alpha was also completely inhibited by neutralizing antibodies specific for ANGII (P < 0.01), but isotype-matched nonimmune immunoglobulins had no significant effect. These data indicate that the induction of AEC apoptosis by TNF-alpha requires a functional renin/angiotensin system (RAS) in the target cell. They also suggest that therapeutic control of AEC apoptosis in response to TNF-alpha is feasible through pharmacologic manipulation of the local RAS.