Predictive profiling of gram-negative antibiotics in CagA oncoprotein inactivation: a molecular dynamics simulation approach.

Gastric cancer (GC) is the fifth most prevalent form of cancer worldwide. CagA - positive Helicobacter pylori infects more than 60% of the human population. Moreover, chronic infection of CagA-positive H. pylori can directly affect GC incidence. In the current study, we have repurposed FDA-approved antibiotics that are viable alternatives to current regimens and can potentially be used as combination therapy against the CagA of H. pylori. The 100 FDA-approved gram negative antibiotics were screened against CagA protein using the AutoDock 4.2 tool. Further, top nine compounds were selected based on higher binding affinity with CagA. The trajectory analysis of MD simulations reflected that binding of these drugs with CagA stabilizes the system. Nonetheless, atomic density map and principal component analysis also support the notion of stable binding of antibiotics to the protein. The residues ASP96, GLN100, PRO184, and THR185 of compound cefpiramide, doxycycline, delafloxacin, metacycline, oxytetracycline, and ertapenem were involved in the binding with CagA protein. These residues are crucial for the CagA that aids in entry or pathogenesis of the bacterium. The screened FDA-approved antibiotics have a potential druggability to inhibit CagA and reduce the progression of H. pylori borne diseases.

Clinical determinants and impact of hemorrhagic lesions on intestinal pathology in preterm infants with surgical necrotizing enterocolitis.

OBJECTIVE: We sought to determine the clinical and histopathological factors associated with intestinal hemorrhage and its correlation with clinical outcomes in neonates with surgical necrotizing enterocolitis (NEC). METHODS: A retrospective study compared clinical and histopathology information in neonates following surgical NEC with severe hemorrhage and those with mild/moderate hemorrhagic lesions seen on resected intestine pathology. RESULTS: The infants with severe hemorrhage (Grade 3-4, 81/148, 54.7%) had significantly lower exposure to antenatal steroids (52.5 % vs 76.9 %; p = 0.004), had higher gestational age (28.5 weeks [7.14] vs. 26.58 [2.90]; p = 0.034), lost more bowel length (p = 0.045), had higher CRP levels at 2 weeks (p = 0.035), and had less intestinal failure ([30.3 % vs 52.5 %]; p = 0.014) than mild/moderate (Grade 0-2, 67/148, 45.2%) hemorrhage group. Those with severe hemorrhage had significantly higher mean inflammation score (2.67 [0.94] vs. 1.63 [0.92]; p = <0.001), higher necrosis scores (1.95 [1.28] vs. 1.49 [1.35]; p = 0.037), higher neovascularization (p = 0.01), higher fibroblasts (p = 0.023) and higher lymphocyte percentages up to 48 hours (p < 0.05) following NEC than mild/ moderate hemorrhage group.On multivariable regression, less exposure to antenatal steroids (OR 0.18 [95% CI 0.05-0.58]; p = 0.005), higher inflammation (OR 3.7 [95% CI 2.09-7.32]; p = 0.001), and lymphocyte count on the day of onset/24 hours following NEC (OR 1.06 [95% CI 1.02-1.11]; p = 0.005) were independently associated with a higher odd of severe intestinal hemorrhage. CONCLUSION: The surgical NEC infants with intestinal hemorrhage were less likely to have antenatal steroid exposure but had higher inflammation grade and lymphocyte counts following NEC onset on multivariable regression modeling.

Clinical outcomes and gestational age based prediction of pneumatosis intestinalis in preterm infants with necrotizing enterocolitis.

BACKGROUND: To determine the clinical implications and gestation age-specific diagnostic predictability of pneumatosis in preterm infants with necrotizing enterocolitis (NEC). METHODS: A retrospective study on abdominal radiographs comparing clinical and radiological information in infants with and without pneumatosis. RESULT: Our findings disproved our hypothesis. Pneumatosis was seen more frequently in infants with higher gestational age [28.4 (26.1-32.4) vs. 26.4 (24.3-29) weeks; p < 0.001] and birth weight [1110 (762-1768) vs. 770 (645-1022) grams; p < 0.001] and were more likely delivered vaginally (39.1% vs. 21.7%, p = 0.01). Portal venous gas was seen frequently on radiographs (10.3% vs. 0%, p < 0.001), but not pneumoperitoneum (20.7% vs. 36.7%, p = 0.02). Infants with pneumatosis frequently developed acute kidney injury, with higher serum creatinine (16.5% vs. 4.5%, p = 0.02) and frequent oliguria (12.9% vs. 2.7 %; p = 0.043) and had higher C-reactive protein levels at 24 and 96 hours (p < 0.002). Receiver operating curves for pneumatosis showed GA >28 weeks and birth weight > 1000 gm to have a sensitivity of 58.6% and specificity of 72.5%. CONCLUSION: Contrary to our hypothesis, infants who developed pneumatosis during NEC were more mature with a higher gestational age and birth weight than those who did not.

Evaluating suitable internal control genes for transcriptional studies in heat-stressed mammary explants of buffaloes.

It is now a well-accepted notion that each new experimental design requires proper evaluation of internal control genes (ICGs) for accurate normalization of expression data. In riverine buffaloes, till date no appropriate ICG has been reported for studying transcriptional response under any of the physiological stressful condition. The objective here was to test 16 well-known reference genes from different functional categories that could serve as suitable ICG during heat stress studies in buffalo mammary tissue. Briefly, the mammary explants were exposed to 45°C for 1 h and subsequently allowed to recover at 37°C for different time points (2-24 h). Three software programs, geNorm, Normfinder and BestKeeper, were used to measure gene transcript stability. RPL22 was excluded because of weak amplification and unacceptable PCR efficiency. Except GAPDH, all other genes showed expression stability within the acceptable range (<1.5). RPL4, B2M, RPS23 and EEF1A1 genes were found to be most stably expressed while GAPDH and ACTB showed least stability. The BestKeeper analysis identified high correlation for RPL4 (r=0.953) and EEF1A1 (r=0.914) with BestKeeper index. Based on the present findings, it could be suggested that geometric average of RPL4, B2M, RPS23 and EEF1A1 would provide accurate normalization to transcriptional data of buffalo mammary explant in response to heat stress.

Selection of suitable reference genes for quantitative gene expression studies in milk somatic cells of lactating cows (Bos indicus).

We assessed the suitability of 9 internal control genes (ICG) in milk somatic cells of lactating cows to find suitable reference genes for use in quantitative PCR (qPCR). Eighteen multiparous lactating Sahiwal cows were used, 6 in each of 3 lactation stages: early (25 ± 5 d in milk), mid (160 ± 15 d in milk), and late (275 ± 25 d in milk) lactation. Nine candidate reference genes [glyceraldehyde 3-phosphate dehydrogenase (GAPDH), protein phosphatase 1 regulatory subunit 11 (PPP1R11), ß-actin (ACTB), ß-2 microglobulin (B2M), 40S ribosomal protein S15a (RPS15A), ubiquitously expressed transcript (UXT), mitochondrial GTPase 1 (MTG1), 18S rRNA (RN18S1), and ubiquitin (UBC)] were evaluated. Three genes, ß-casein (CSN2), lactoferrin (LTF), and cathelicidin (CAMP) were chosen as target genes. Very high amplification was observed in 7 ICG and very low level amplification was observed in 2 ICG (UXT and MTG1). Thus, UXT and MTG1 were excluded from further analysis. The qPCR data were analyzed by 2 software packages, geNorm and NormFinder, to determine suitable reference genes, based on their stability and expression. Overall, PPP1R11, ACTB, UBC, and GAPDH were stably expressed among all candidate reference genes. Therefore, these genes could be used as ICG for normalization of qPCR data in milk somatic cells through lactation.

A factor analytic and psychometric examination of pathology of separation-individuation.

Two studies are described that attempt to determine if standard-scale-reduction techniques could yield a construct-valid diagnostic screen of pathology of separation-individuation for use in nonclinical university settings. In Study 1 (N = 210), a measure of pathology of separation-individuation (PATHSEP) was reduced successfully to a single, internally consistent factor, accounting for 36% of the variance. In Study 2 (N = 304), these items also coalesced around a single factor, accounting for 35% of the variance. Study 2 also showed that PATHSEP is correlated moderately and positively with indices of insecure attachment, with the Center for Epidemiological Studies-Depression Scale, and with indices of psychiatric symptomatology (Hopkins Symptom Checklist). PATHSEP also was associated with a poorer profile of adjustment to college. Males reported more pathology of separation-individuation than did females. Evidence supports the construct validity of a shortened version of PATHSEP. Directions for future research are noted.

Quantitative analysis of hydrocortisone in human urine using a high-performance liquid chromatographic-tandem mass spectrometric-atmospheric-pressure chemical ionization method.

In this study, the development and validation of a method of analysis for 11,17,21,-trihydroxypregn-4-ene-3,20-dione (hydrocortisone, cortisol, HC) using high-performance liquid chromatography (HPLC)-tandem mass spectrometry (MS) with atmospheric-pressure chemical ionization (APCI) is reported. This is the first report of the systematic development and validation of an HPLC-MS-MS method for the quantitation of HC in synthetic human urine with a deuterated internal standard. Prior to LC-MS-MS analysis, the only sample preparation used was the dilute-and-shoot technique prior to LC-MS-MS analysis. In this study, an analysis time of less than 3 min is achieved. The results show freedom of interference from other analytes such as analogous steroids. Validation parameters such as specificity/selectivity, limit of quantitation (LOQ), linearity, precision, accuracy, ruggedness, stability, and system suitability are evaluated for this method. The LOQ is 5 ng/mL with an 8% relative standard deviation (RSD). For calibration standard curves, an average linear response for a 3-day validation (R2 = 0.997) over the range of 5 to 500 ng/mL is obtained. The interday precision %RSDs are 7.2, 5.0, and 5.2 for 15, 75, and 300 ng/mL, respectively. Also, brief comparisons of the dilute-and-shoot and liquid-liquid extraction techniques for this analyte are discussed.

Pathological attachment and attachment style in late adolescence.

Attachment theory is frequently invoked to account for patterns of adaptation within relationships. West and Sheldon derived a measure of dysfunctional adult attachment from Bowlby's theory. Four patterns are identified: compulsive self-reliance; caregiving; care-seeking; and angry withdrawal. The aim of this study was to assess the psychometric properties of this promising measure, and to assess its ability to predict symptomatology relative to measures of attachment style. Participants included 209 late adolescents who were involved in committed relationships. They responded to categorical and continuous measures of attachment style and various measures of symptomatology and college adjustment. The four dimensions of pathological attachment showed strong internal consistency and few gender differences. Compulsive care-seeking, angry withdrawal and compulsive self-reliance were particularly strong predictors of psychiatric symptomatology and insecure attachment style. Attachment pathology and insecure attachment style predicted symptomatology similarly, though variations were evident depending on how style was assessed. This study is the first to provide evidence of convergent, discriminant and predictive validity for this measure. Directions for future research are noted.

In vivo effect of cyanate on serum and eye lens in rat.

Supplementation of cyanate in rats caused a significant decrease in serum GSH and increase in calcium and phosphate level both in serum and lens. Consequently, these changes led to induce acidosis uremia in serum and hypercalcemia and hyperphosphatemia in lens which may be possible causing factor for cataract.

Role of trace elements in senile cataract.

The contents of copper, zinc and protein were determined in cortex and nucleus sections of cortical and nuclear cataracts. The results show that copper and zinc content increased in both types of cataracts, although more in the cortex than nucleus sections of the cataractous lenses.

Clinical apnea as an early manifestation of Pneumocystis carinii pneumonia in an infant with perinatal HIV-1 infection.

This article describes the first case of Pneumocystis carinii pneumonia in a human immunodeficiency virus (HIV) seropositive infant in whom apnea was the earliest presenting clinical finding. Pediatricians treating infants with HIV infection need to be aware of this unusual clinical presentation of P carinii pneumonia to avoid a delay in diagnosis and management of this disease.