Nuclear Vav3 is required for polycomb repression complex-1 activity in B-cell lymphoblastic leukemogenesis.

Acute B-cell lymphoblastic leukemia (B-ALL) results from oligo-clonal evolution of B-cell progenitors endowed with initiating and propagating leukemia properties. The activation of both the Rac guanine nucleotide exchange factor (Rac GEF) Vav3 and Rac GTPases is required for leukemogenesis mediated by the oncogenic fusion protein BCR-ABL. Vav3 expression becomes predominantly nuclear upon expression of BCR-ABL signature. In the nucleus, Vav3 interacts with BCR-ABL, Rac, and the polycomb repression complex (PRC) proteins Bmi1, Ring1b and Ezh2. The GEF activity of Vav3 is required for the proliferation, Bmi1-dependent B-cell progenitor self-renewal, nuclear Rac activation, protein interaction with Bmi1, mono-ubiquitination of H2A(K119) (H2AK119Ub) and repression of PRC-1 (PRC1) downstream target loci, of leukemic B-cell progenitors. Vav3 deficiency results in de-repression of negative regulators of cell proliferation and repression of oncogenic transcriptional factors. Mechanistically, we show that Vav3 prevents the Phlpp2-sensitive and Akt (S473)-dependent phosphorylation of Bmi1 on the regulatory residue S314 that, in turn, promotes the transcriptional factor reprogramming of leukemic B-cell progenitors. These results highlight the importance of non-canonical nuclear Rho GTPase signaling in leukemogenesis.

Evaluation of spatial spreading of phyto-available sulphur and micronutrients in cultivated coastal soils.

Understanding the spatial spreading patterns of plant-available sulphur (S) (AS) and plant-available micronutrients (available zinc (AZn), available iron (AFe), available copper (ACu), available manganese (AMn) and available boron (AB)) in soils, especially in coastal agricultural soils subjected to various natural and anthropogenic activities, is vital for sustainable crop production by adopting site-specific nutrient management (SSNM) strategies. We studied the spatial distribution patterns of AS, AZn, AFe, ACu, AMn, and AB in cultivated soils of coastal districts of India using geostatistical approaches. Altogether 39,097 soil samples from surface (0 to 15 cm depth) layers were gathered from farm lands of 68 coastal districts. The analysis of soil samples was carried out for soil pH, electrical conductivity (EC), soil organic carbon (SOC) and AS, AZn, AFe, ACu, AMn, and AB. Soil pH, EC and SOC varied from 3.70 to 9.90, 0.01 to 7.45 dS m-1 and 0.02 to 3.74%, respectively. The concentrations of AS, AZn, AFe, ACu, AMn, and AB varied widely in the study area with their corresponding mean values were 37.4±29.4, 1.50±1.53, 27.9±35.1, 2.14±1.74, 16.9±18.4 and 1.34±1.52 mg kg-1, respectively. The coefficient of variation values of analyzed soil parameters varied from 14.6 to 126%. The concentrations of AS, AZn, AFe, ACu, AMn, and AB were negatively and significantly correlated with soil pH and positively and significantly correlated with SOC. The geostatistical analysis indicated stable, Gaussian and exponential best-fit semivariogram models with moderate to strong spatial dependence for available nutrients. The generated spatial spreading maps revealed different distribution patterns for AS, AZn, AFe, ACu, AMn, and AB. There were variations in spatial spreading patterns of AS, AZn, AFe, ACu, AMn, and AB in east- and west-coastal area. About 62, 35, 12, 0.4, 23 and 45% of the study area had deficiency of AS, AZn, AFe, ACu, AMn, and AB, respectively. The spatial spreading maps will be highly useful for SSNM in the cultivated coastal soils of the country. This study could also be used as a base for assessing spatial spreading patterns of soil parameters in cultivated coastal areas of other parts of the world.

Persistence of marine heat waves for coral bleaching and their spectral characteristics around Andaman coral reef.

Coral reefs are fragile and endangered ecosystems in the tropical marine and coastal environment. Thermal stress due to marine heat waves (MHW) could cause significantly negative impacts on the health conditions, i.e., bleaching of the coral ecosystem. The current study is an attempt to quantify the intensity of coral bleaching in the Andaman region in recent decades using the intensity of marine heat wave (IMHW) estimated from satellite measured sea surface temperature (SST). A linear regression model was developed between IMHW and in situ observations of percent coral bleaching (PCB) which has the slope 7.767 (of IMHW unit) and intercept (- 141.7). Further, an attempt was also made to establish the relationship between PCB and the ratio between the remote sensing reflectance (Rrs) at 443 and 531 nm to upscale the percentage of coral bleaching at synoptic scales. A significant positive correlation between the PCB and band ratio index was found (R2 = 0.72). This approach can be used for the operational monitoring of coral reef beaching in this region.

The effect of aloe vera and tea tree oil mouthwashes on the oral health of school children.

PURPOSE: To evaluate the effect of two herbal mouthwashes containing aloe vera and tea tree oil, on the oral health of school children. METHODS: A double-blinded, placebo-controlled prospective interventional study was conducted in school children aged 8-14 years. The study participants were divided into four groups depending upon the mouthwash used: Group 1 (aloe vera), Group 2 (chlorhexidine), Group 3 (tea tree oil) and Group 4 (placebo). The variables studied included plaque index, gingival index and salivary Streptococcus mutans counts, which were recorded at baseline, 4 weeks after supervised mouth rinse and after 2 weeks of stopping the mouth rinse. RESULTS: A total of 89 boys and 63 girls were included. A statistically significant decrease in all variables was noted after the use of both the herbal preparations at the end of 4 weeks which was maintained after the 2-week washout period (p < 0.001). The difference in variables between groups using aloe vera, Tea tree oil and chlorhexidine, was not statistically significant. CONCLUSION: The use of aloe vera and tea tree oil mouthwashes can decrease plaque, gingivitis and S. mutans in the oral cavity in children. The activity of these two agents is comparable to that of chlorhexidine.

Comparison of histochemical staining techniques for detecting mast cells in oral lesions.

Mast cells are large cells with granular cytoplasm that participate in wound healing, angiogenesis and defense against pathogens. They also contribute to inflammation by initiating innate and acquired immunity. The granules of these cells exhibit characteristic staining properties. We investigated toluidine blue, astra blue, Alcian blue-pyronin Y and May-Grunwald Giemsa stains for mast cells in various oral lesions and assessed the efficacy of each for identifying mast cells. Sections were obtained from 10 each of diagnosed cases of inflammatory fibrous hyperplasia, periapical cyst, mild dysplasia, oral submucous fibrosis and oral squamous cell carcinoma and stained using the stains listed above. Mast cells were assessed for their presence, contrast of the mast cell in the connective tissue background and number. We found that May-Grunwald Giemsa stain was the best for identification of mast cells, although toluidine blue staining is less time-consuming. Overall we obtained better results using May-Grunwald Giemsa and toluidine blue for staining mast cells.

Development and evaluation of retort pouch processed chhenapoda (cheese based baked sweet).

A study has been undertaken to optimize ingredient proportions for preparation of chhenapoda and the effect of retort processing on its quality and storability. Chhenapoda was prepared from cottage cheese with standard practices followed by confectioners using different levels of semolina (0, 50, 100, 150, 200 g) and sugar (0, 100, 200, 300, 400 g) per kg cheese and ingredient proportion was optimized based on sensory scores. Prepared chhenapoda sample of 200 g were packed in pre-fabricated 3 side seal multilayer laminated retortable pouches, hermetically sealed and retort processed at different temperatures (100, 110 and 120 °C) and time intervals (15, 30 and 45 min). It was found from Response Surface Methodology (RSM) technique that addition of 18.5% sugar and 7.5% semolina with cottage cheese was optimum for chhenapoda preparation. Microbiological analysis showed that total plate count and yeast and mould count (YMC) decreased from 110 × 107 to 4 × 104 and YMC from 3 × 103 to 0 respectively during retort processing (30 min thermal processing in laminated pouch at 120 °C). From the storage study, it can be concluded that retort processing of chhenapoda in laminated pouch at 120 °C for 30 min resulted in microbiological safe and sensory acceptable product which could be stored up to 30 days under refrigerated condition.

The signaling axis atypical protein kinase C λ/ι-Satb2 mediates leukemic transformation of B-cell progenitors.

Epigenetically regulated transcriptional plasticity has been proposed as a mechanism of differentiation arrest and resistance to therapy. BCR-ABL leukemias result from leukemic stem cell/progenitor transformation and represent an opportunity to identify epigenetic progress contributing to lineage leukemogenesis. Primary human and murine BCR-ABL+ leukemic progenitors have increased activation of Cdc42 and the downstream atypical protein kinase C (aPKC). While the isoform aPKCζ behaves as a leukemic suppressor, aPKCλ/ι is critically required for oncogenic progenitor proliferation, survival, and B-cell differentiation arrest, but not for normal B-cell lineage differentiation. In vitro and in vivo B-cell transformation by BCR-ABL requires the downregulation of key genes in the B-cell differentiation program through an aPKC λ/ι-Erk dependent Etv5/Satb2 chromatin repressive signaling complex. Genetic or pharmacological targeting of aPKC impairs human oncogenic addicted leukemias. Therefore, the aPKCλ/ι-SATB2 signaling cascade is required for leukemic BCR-ABL+ B-cell progenitor transformation and is amenable to non-tyrosine kinase inhibition.

Assessment of Cognitive Impairment in Patients with Chronic Obstructive Pulmonary Disease Using the Rapid Cognitive Screen.

OBJECTIVES: To determine the prevalence of cognitive impairment in chronic obstructive pulmonary disease (COPD) patients using the Rapid Cognitive Screen (RCS). DESIGN: Cross sectional study. PARTICIPANTS: A total of 106 men and women with COPD 50 years of age or older. SETTING: Pulmonary ambulatory clinic in a tertiary academic center in St. Louis, Missouri, USA. MEASUREMENTS: We enrolled patients with COPD who completed the RCS tool. Patients were classified as having normal cognition, mild cognitive impairment (MCI), or dementia if they scored ≥ 8, 6-7, or ≤ 5, respectively. The prevalence of each category was measured, and patients' baseline characteristics were compared. RESULTS: One hundred and six patients completed the RCS. Of those, 36 (33.9%) patients scored ≤ 5 on the RCS, 33 (31.1%) scored 6 or 7, and 37 (34.9%) scored ≥ 8. The prevalence of dementia, MCI, and normal cognition were 33.9%, 31.1%, and 34.9%, respectively. Baseline lung function was not different between patients with normal cognition and those with abnormal cognition (FEV1 1.53 ± 0.39 vs 1.45 ± 0.32 L). In patients with abnormal cognition, there was no difference in patients' characteristics between patients with MCI or dementia. CONCLUSION: Cognitive impairment is highly prevalent in patients with COPD. RCS successfully identified cognitive impairment in COPD. We recommend performing RCS in patients with COPD routinely.

Temperature Controlled Evolution of Silver Polypods from Biomass Encapsulated Nano-Droplets.

Silver nanoparticles are produced by a novel biosynthesis technique using fungus Penicillium purpurogenum NPMF (MTCC 7356). The biomass when subjected to heat treatment up to 700 °C produces the nanoparticles of silver due to the presence of free carbon which acts as encapsulation for silver nanoparticles. Increase in temperature above 700 °C causes a reduction in concentration of free carbon so that the nanoparticles come close to each other and form clusters in the shape of tetrapods to polypods. The micro-Raman analysis supports the formation of mesoporous carbon with increase in the calcined temperature. The formation of Ag nanostructures has been explained in terms of free energy diagram i.e., the crystal favors more to grow on (100) facet or (110) facet rather than on (111) facet by the addition of silver atoms and thereby, it releases more energy. The release of energy has been confirmed from the differential thermo gravimetric analysis. In fact, it is the dynamics of the minimization of the free energy of the system which governs the reorientation and rearrangement of Ag themselves leading to formation of various types of polypods. A formation mechanism is presented schematically.

Immunohistochemical localization of TLR2 and CD14 in gingival tissue of healthy individuals and patients with chronic periodontitis.

We used immunohistochemistry to quantify and compare the expression of Toll-like receptor 2 (TLR2) and cluster of differentiation 14 (CD14) in gingival tissues of both healthy individuals and patients with chronic periodontitis. We also correlated the expression of TLR2 and CD14 with the histological grades of chronic periodontitis. We examined 30 gingival specimens from chronic periodontitis patients and 10 from healthy individuals. Tissues from both groups were immunostained with antibodies against TLR2 and CD14. TLR2 and CD14 were expressed by endothelial cells, fibroblasts, lymphocytes and plasma cells. The immunohistochemical expression of TLR2 and CD14 was significantly greater in inflammatory cells of the chronic periodontitis group than in healthy individuals. Expression of these molecules was greater in the inflammatory cells of connective tissue adjacent to pocket epithelium in both groups. The expression of TLR2 and CD14 was greatest in the periodontitis group that was classified as severe grade, followed by moderate and mild grades, which suggests a role of TLR2 and CD14 in the pathogenesis of chronic periodontitis. The positive correlation of TLR2 and CD14 expression levels with the severity grades of chronic periodontitis suggests that they are correlated also with disease severity; therefore, they may be useful for predicting disease progression. Our findings are consistent with the possibility that CD14 acts as a co-receptor for TLR2.

Sexual Dysfunction among Females Receiving Psychotropic Medication: A Hospital-based Cross-sectional Study.

BACKGROUND: Sexual dysfunction (SD) is a known adverse effect of psychotropic medications. Even though sexual difficulties are common among women; very few studies have been carried out in India. OBJECTIVE: To study the prevalence and nature of SD among females receiving psychotropic medications and to compare the SD among female patients receiving antipsychotics and antidepressants. MATERIALS AND METHODS: Female investigator conducted a hospital-based cross-sectional study on female patients visiting the psychiatry outpatient department. Patients meeting inclusion criteria were assessed for SD disorder as per Diagnostic and Statistical Manual of Mental Disorders 4th Edition Text Revision. SD severity was measured using Female Sexual Function Index (FSFI) scale. RESULTS: The prevalence of SD in this study was 68.32%. There was more than one SD in 48 (47.52%). FSFI score was significantly low in patients with SD as compared to patients not having SD (P = 0.001). SD was more common in patients who were on combination of antidepressants and benzodiazepines than antidepressant alone or antipsychotic alone. CONCLUSION: SD was prevalent in more than 50% of female patients on psychotropic drugs. Number of patients on individual psychotropic drugs was so small that a definite conclusion could not be drawn. Study emphasizes the need to carry out similar study on larger number of patients to get better insight into this problem.

A histochemical comparison of methyl green-pyronin, and hematoxylin and eosin for detecting apoptotic cells in oral squamous cell carcinoma, oral leukoplakia, oral submucous fibrosis and normal oral mucosa.

Analysis of apoptotic cells in oral pathological states could be useful for determining the rates of tissue turnover, which would help determine prognosis. The use of histochemical stains such as hematoxylin and eosin (H & E) and methyl green-pyronin (MGP) can provide a simple and cost-effective method for detecting apoptotic cells. We compared the efficacy of MGP and H & E for detecting apoptotic cells in oral squamous cell carcinoma (OSCC), oral leukoplakia (OL), oral submucous fibrosis (OSMF) and normal oral mucosa (NOM). Ten cases each of OSCC, OSMF, OL and NOM were retrieved from the archives and two serial sections were stained, one with H & E and the other with MGP. Apoptotic cells were identified at 100 x magnification and the apoptotic index was calculated. Apoptotic cells were distinguished more readily in MGP stained sections than in those stained with H & E. Also, the apoptotic cell count was greater in OSCC compared to OL, OSMF and NOM. We concluded that MGP staining can be used as a routine, cost-effective method for detecting apoptotic cells.

Hoover's sign: Clinical relevance in Neurology.

Hoover's sign was described by Dr. Charles Franklin Hoover more than 100 years back to differentiate between the organic and functional weakness of pyramidal origin. This test is usually performed in the lower limbs and is valuable when on bedside one is not sure about the nature of hemiparesis. A subject with hemiparesis of organic cause while asked to flex the hip of normal leg against resistance will not exert pressure on the hand of examiner placed under the heel on the affected side while in hysterical weakness heightened pressure will be felt on the examiner's hand. The presumed genesis of this sign could be the crossed extensor reflex or the principle of synergistic contraction. It is a useful clinical test in differentiating functional and organic paresis with moderate sensitivity (63%) and high specificity (100%), but there are some limitations which should be kept in mind while evaluating a patient.

PCR based detection of HPV 16 and 18 genotypes in normal oral mucosa of tobacco users and non-users.

There is increasing evidence of a causal association between human papillomavirus (HPV) and oral squamous cell carcinoma (OSCC). Several studies have shown that HPV is associated with increased risk of oral cancer independent of exposure to tobacco and alcohol. The association is valid for HPVs 16 and 18, which generally are considered high risk types, because they have been detected in oral dysplastic lesions and cancers. We determined the baseline prevalence of HPVs 16 and 18 in normal oral mucosa of individuals with and without tobacco habit. PCR was used for DNA collected by oral smears to detect HPV 16/18 DNA in normal oral mucosa of 60 healthy individuals who were assigned to two groups of 30 subjects each. One group had a tobacco habit, the other did not. The tobacco user group comprised individuals who were tobacco chewers only. Sixty-five percent of individuals were positive for HPV 16/18 DNA, but HPV 16/18 positivity was less in individuals with tobacco habit than in those without tobacco habit. No significant association was found between the presence of HPVs and gender, age or duration of chewing habit, or between groups with and without a tobacco habit. We propose that HPVs16 and 18 commonly are present in normal oral mucosa and emphasize the importance of distinguishing clinical, subclinical and latent HPV infections when investigating HPVs and OSCC.

Accuracy of spot urine protein creatinine ratio in measuring proteinuria in chronic kidney disease stage 3 and 4.

We studied the accuracy of spot urine protein creatinine ratio (SpUr-PCR) to assess 24 h urine protein excretion (24 h-UP) in patients with chronic kidney disease (CKD). A total of 100 proteinuric CKD patients of stages 3 and 4 were studied. 24 h urine was collected to measure 24 h-UP and creatinine. A random day time urine sample was analyzed to measure the PCR. A formula to estimate 24 h creatinine excretion was derived from linear regression analysis and a correction factor was introduced to assess whether this improves the accuracy of the SpUr PCR in predicting 24 h-UP. Accuracy of the SpUr-PCR was assessed by Pearson's correlation, regression analysis, and Bland Altman analysis. Mean age was 51.85 ± 12 years and 81% of the patients were male. SpUr-PCR predicted 24 h-UP with good accuracy (r = 0.86 on a data transformed to a logarithmic scale, P < 0.001) and there was a good agreement between these two measures of proteinuria. However, SpUr-PCR was inaccurate in the subgroup with nephrotic range proteinuria (r = 0.35, P = 0.062), but when a correction factor for 24-h urine creatinine (24 h-UCr) was introduced, the accuracy of SpUr-PCR improved significantly in this group (r = 0.45, P = 0.013). Introduction of the correction factor improved the degree of agreement between these two measures in women, but not the correlation. Overall, SpUr-PCR accurately predicted 24 h-UP. Adding a correction factor for 24 h-UCr improved correlation in the subgroup of patients with the nephrotic range proteinuria and the degree of agreement in female patients, and hence may be used in expressing proteinuria measured by SpUr-PCR to improve its accuracy in them.

Attenuation of oxidative hemolysis of human red blood cells by the natural phenolic compound, allylpyrocatechol.

The protecting ability of the Piper betle leaves-derived phenol, allylpyrocatechol (APC) against AAPH-induced membrane damage of human red blood cells (RBCs) was investigated. Compared to control, AAPH (50 mM) treatment resulted in significant hemolysis (55%, p < 0.01), associated with increased malondialdehyde (MDA) (2.9-fold, p < 0.001) and methemoglobin (6.1-fold, p < 0.001) levels. The structural deformation due to membrane damage was confirmed from scanning electron microscopy (SEM) images and Heinz bodies formation, while the cell permeability was evident from the K(+) efflux (28.7%, p < 0.05) and increased intracellular Na(+) concentration (8%, p < 0.05). The membrane damage, due to the reduction of the cholesterol/phospholipids ratio and depletion (p < 0.001) of ATP, 2,3-DPG by ˜44-54% and Na(+)-K(+) ATPase activity (43.7%), indicated loss of RBC functionality. The adverse effects of AAPH on all these biochemical parameters and the resultant oxidative hemolysis of RBCs were significantly reduced by pretreating the cells with APC (7 µM) or α-tocopherol (50 µM) for 1 h, prior to incubation with AAPH.

Immunohistochemical expression of TLR4 and TLR9 in various grades of oral epithelial dysplasia and squamous cell carcinoma, and their roles in tumor progression: a pilot study.

Toll-like receptors (TLRs) play an essential role in the activation of innate immunity. TLRs are expressed in B-lymphocytes, monocytes, dendritic cells and epithelial cells. We examined the immunohistochemical expressions of TLR4 and TLR9 in various grades of oral epithelial dysplasia (OED), oral squamous cell carcinoma (OSCC) and normal oral mucosa (NOM) to determine the association between TLR4 and TLR9 in the progression of lesions from dysplasia to carcinoma. Expressions of TLR4 and TLR9 were assessed using immunohistochemistry (IHC) on paraffin embedded tissue blocks of various grades of OED (28 cases), OSCC (27 cases) and NOM (10 cases). Expression of TLR4 was high in all grades of OED and OSCC. Expression of TLR9 was high in well differentiated squamous cell carcinoma and moderately differentiated squamous cell carcinomas, and moderate to low in poorly differentiated squamous cell carcinomas. Although expression was high in case of TLR4, it was not statistically significant. Expression of TLR9 was statistically significant. In OED, expression of TLR9 was less than that of TLR4. Our results indicated that the pattern of expression of TLR4 and TLR9 increased significantly from mild to severe dysplasia compared to controls. Expression of TLR4 and TLR9 reflects progression of OED to OSCC, which suggests that TLR may play a role in tumorigenesis and that it could be used as a target for OSCC prevention and therapy in the future.

Klf5 controls bone marrow homing of stem cells and progenitors through Rab5-mediated ß1/ß2-integrin trafficking.

Krüppel-like factor 5 regulates pluripotent stem cell self-renewal, but its role in somatic stem cells is unknown. Here we show that Krüppel-like factor 5-deficient haematopoietic stem cells and progenitors fail to engraft after transplantation. This haematopoietic stem cell and progenitor defect is associated with impaired bone marrow homing and lodging and decreased retention in bone marrow, and with decreased adhesion to fibronectin and expression of membrane-bound ß1/ß2-integrins. In vivo-inducible gain-of-function of Krüppel-like factor 5 in haematopoietic stem cells increases haematopoietic stem cell and progenitor adhesion. The expression of Rab5 family members, mediators of ß1/ß2-integrin recycling in the early endosome, is decreased in Klf5(Δ/Δ) haematopoietic stem cells and progenitors. Krüppel-like factor 5 binds directly to the promoter of Rab5a/b, and overexpression of Rab5b rescues the expression of activated ß1/ß2-integrins, adhesion and bone marrow homing of Klf5(Δ/Δ) haematopoietic stem cells and progenitors. Altogether, these data indicate that Krüppel-like factor 5 is indispensable for adhesion, homing, lodging and retention of haematopoietic stem cells and progenitors in the bone marrow through Rab5-dependent post-translational regulation of ß1/ß2 integrins.