Molecular detection of the G(-248)A BAX promoter nucleotide change in B cell chronic lymphocytic leukaemia.

BACKGROUND: A novel single nucleotide polymorphism (SNP), G(-248)A, in the 5' untranslated region of the BAX promoter and its association with reduced protein expression, progression beyond Rai stage 0, and treatment resistance in chronic lymphocytic leukaemia (CLL) has been reported previously. AIM: To develop a restriction enzyme analysis (REA) based method for routine detection of BAX promoter SNP in a clinical laboratory. METHODS: The BAX promoter was analysed in duplicate by REA and sequencing in 90 samples (from 45 patients with CLL, 43 controls, and two cell lines). The promoter region was amplified, digested with restriction endonucleases (Aci I and Tau I), and separated by gel electrophoresis. RESULTS: After digestion, the normal GG genotype samples produced three distinct bands. The homozygous AA replacement abolished the cleavage site, resulting in a single band. Although the heterozygous samples produced three bands, the two smaller visible bands were reduced in intensity (> 50%). The test characteristics of Aci I REA were better than those of Tau I REA, in terms of sensitivity (100% v 77.8%), specificity (98.6% v 92.3%), positive predictive value (95.03% v 87.4%), and negative predictive value (100% v 85.83%). CONCLUSIONS: REA using Aci I is a highly sensitive and specific method for detecting the BAX G(-248)A SNP in CLL.     

Involvement of Insulin-Like Growth Factor 1 Receptor Signaling in the Amyloid-β Peptide Oligomers-Induced p75 Neurotrophin Receptor Protein Expression in Mouse Hippocampus

The p75 neurotrophin receptor (p75NTR) has been thought to play a critical role in amyloid-β peptide (Aβ)-mediated neurodegeneration and Aβ metabolism in Alzheimer's disease (AD) brains. Our previous report showed that membrane-associated p75NTR protein expression was significantly increased in the hippocampi of two different strains of transgenic AD mice and was associated with the age-dependent elevation of Aβ1-42 levels. Here, we provide evidence that the Aβ1-42 oligomers known as ADDLs (Aβ-derived diffusible ligands) induce p75NTR protein expression through insulin-like growth factor 1 receptor (IGF-1R) phosphorylation in SH-SY5Y human neuroblastoma cells. An in vivo microinjection study demonstrated that microinjected ADDLs increased the p75NTR protein expression by 1.4-fold in the ipsilateral hippocampus compared to the contralateral hippocampus. In addition, ADDLs microinjected into mouse hippocampi facilitated IGF-1R phosphorylation within 30 min and the co-administration of picropodophyllin, an IGF-1R kinase inhibitor, blocked ADDLs-induced p75NTR expression. We examined the possible involvement of IGF-1R in the increased p75NTR protein expression in the hippocampi of 6-month-old AβPPswe/PS1dE9 AD model mice that had accumulated significant amounts of Aβ1-42 and showed significantly higher p75NTR expression than age-matched wild-type mice. We found that IGF-1R phosphorylation in these transgenic mice was higher than that in the wild-type mice. These findings indicate that Aβ1-42 oligomers stimulate the p75NTR protein expression in the hippocampus through IGF-1R signaling. Thus, Aβ1-42 oligomers-mediated IGF-1R activation may trigger an increase in p75NTR protein expression in the hippocampus of AD brain during the early stages of disease development.     

PRogram In Support of Moms (PRISM): a pilot group randomized controlled trial of two approaches to improving depression among perinatal women

Purpose: This pilot study was designed to inform a larger effectiveness trial by: (1) assessing the feasibility of the PRogram In Support of Moms (PRISM) and our study procedures; and, (2) determining the extent to which PRISM as compared to an active comparison group, the Massachusetts Child Access Psychiatry Program (MCPAP) for Moms alone, improves depression among perinatal women.

Methods: Four practices were randomized to either PRISM or MCPAP for Moms alone, a state-wide telephonic perinatal psychiatry program. PRISM includes MCPAP for Moms plus implementation assistance with local champions, training, and implementation of office prompts and procedures to enhance depression screening, assessment and treatment. Patients with Edinburgh Postnatal Depression Scales (EPDS)?≥?10 were recruited during pregnancy, and completed the EPDS and a structured interview at baseline and 3–12 weeks’ postpartum.

Results: Among MCPAP for Moms alone practices, patients’ (n?=?9) EPDS scores improved from 15.22 to 10.11 (p?=?0.010), whereas in PRISM practices patients’ (n?=?21) EPDS scores improved from 13.57 to 6.19 (p?=?0.001); the between groups difference-of-differences was 2.27 (p?=?0.341).

Conclusions: PRISM was beneficial for patients, clinicians, and support staff. Both PRISM and MCPAP for Moms alone improve depression symptom severity and the percentage of women with an EPDS >10. The improvement difference between groups was not statistically significant due to limited power associated with small sample size.     

Factors Influencing the Varus Deformity of Humeral Head in Proximal Humerus Fractures and Its Relation to Functional Outcome

BackgroundFractures of the proximal humerus represent approximately 4% of all fractures and 26% of humerus fractures. Proper reduction, stable internal fixation and early initiation of physiotherapy help to achieve a good functional outcome. Aim of this study was to evaluate varus fixation/malunion of proximal humerus fractures and its relation to functional outcome.Materials and MethodsWe retrospectively evaluated 32 patients with proximal humerus fractures who were surgically treated between 2015 and 2017 at tertiary care hospital. We divided the patients into three groups on the basis of the neck-shaft angle as valgus group, normal group and varus group to observe the influence of neck-shaft angle on efficacy. Patients were evaluated for functional outcome using the Constant–Murley score.ResultsTwo-part fractures had better functional outcome (Constant score = 75.15) compared to three parts with the moderate functional outcome (Constant score = 68.81) and the four-part fracture had poor functional outcome (Constant score = 52.66). After 6 months of follow-up, 13 patients had a neck-shaft angle of less than 126°. The functional outcome is significantly better among patients with normal neck-shaft angle and had a mean Constant score of 76.63 as compared to patients with varus deformity had a mean Constant score 60 (p = 0.001). 10 patients did not have medial support, in which 08 patients had neck-shaft angle less than 126° and 2 had a normal neck-shaft angle.ConclusionHigh fracture comminution, improper restoration of medial continuity causes varus deformity of the humeral head and it leads to poor functional outcome. The small sample size is the limitation of our study.     

Involvement of nitric oxide synthase and ROS-mediated activation of L-type voltage-gated Ca2+ channels in NMDA-induced DPYSL3 degradation

Dihydropyrimidinase-like 3 (DPYSL3), a member of TUC (TOAD-64/Ulip/CRMP), is believed to play a role in neuronal differentiation, axonal outgrowth and possibly in neuronal regeneration. Recently, we have shown that in primary cortical neurons (PCN) NMDA and oxidative stress (H(2)O(2)) caused a calpain-dependent cleavage of DPYSL3 (62 kDa) resulting in the appearance of a lower molecular weight form (60 kDa) of DPYSL3. Our preliminary results had shown that antioxidants significantly reduced NMDA-induced DPYSL3 degradation, indicating involvement of ROS in calpain activation. The aim of this study was to investigate the possible involvement of NOS in NMDA-induced DPYSL3 degradation. We found that NOS inhibitor (L-NAME) significantly prevented NMDA-induced ROS formation, as well as intracellular Ca(2+) increase [Ca(2+)](i), DPYSL3 degradation and cell death. Further, exposure of PCN to NO donor (SNP) resulted in significant [Ca(2+)](i) increase, ROS generation and probable calpain-mediated DPYSL3 truncation. The NMDA- and oxidative stress (ROS)-induced DPYSL3 truncation was totally dependent on extracellular [Ca(2+)](i). While NMDA-induced DPYSL3 truncation was blocked by both NMDA receptor antagonist (MK801) [Kowara, R., Chen, Q., Milliken, M., Chakravarthy, B., 2005. Calpain-mediated degradation of dihydropyrimidinase-like 3 protein (DPYSL3) in response to NMDA and H(2)O(2) toxicity. J. Neurochem. 95 (2), 466-474] and L-VGCC (nimodipine) inhibitors, H(2)O(2)-induced increase in [Ca(2+)](i), ROS generation and DPYSL3 truncation was blocked only by nimodipine. These results indicate that changes in Ca(2+) homeostasis resulting from ROS-dependent activation of L-VGCC are sufficient to induce probable calpain-mediated DPYSL3 truncation and demonstrate for the first time the role of ROS in the mechanism leading to glutamate-induced calpain activation and DPYSL3 protein degradation. The probable calpain-mediated DPYSL3 truncation may have significant impact on its interaction with actin and its assembly, and in turn on growth cone integrity.     

Modulatory role of grape seed extract on age-related oxidative DNA damage in central nervous system of rats

Aging is the accumulation of diverse deleterious changes in the cells and tissues leading to increased risk of diseases. Oxidative stress is considered as a major risk factor and contributes to age related increase in DNA oxidation and DNA protein cross-links in central nervous system during aging. In the present study, we have evaluated the salubrious role of grape seed extract on accumulation of oxidative DNA damage products such as 8-OHdG and DNA protein cross-links in aged rats. Male albino rats of Wistar strain were divided into four groups: Group I, young control rats; Group II, young rats treated with grape seed extract (100 mg/kg b.wt.) for 30 days; Group III, aged control rats; Group IV, aged rats supplemented with grape seed extract (100 mg/kg b.wt.) for 30 days. Our results, thus, revealed that grape seed extract has inhibiting effect on the accumulation of age-related oxidative DNA damages in spinal cord and in various brain regions such as cerebral cortex, striatum and hippocampus.     

Serum protein profile studies of cervical cancers in monitoring of tumor response to radiotherapy using HPLC-LIF: A pilot study

ObjectiveExploration of the feasibility of serum protein profiles for monitoring tumor radioresponse in cervical cancers using HPLC-LIF system.Materials and methodsTwenty-one subjects were recruited in the study. Out of them 7 were healthy, 14 were cervical cancer patients who undertook fractionated radiotherapy (RT) with 2 Gy per fraction over 25 fractions, for 5 weeks followed by 2 applications of intracavitary brachytherapy once a week.Blood collected from above subjects was processed to obtain serum. Serum chromatograms of ‘normal’ (n=7) and conspicuous probes before RT (n=14, ‘malignant’) and 24 h after second fraction of RT (n=13, ‘2-RT’), were recorded using an In-house-built HPLC-LIF set-up. Data were analyzed in two approaches: (1) classical method using relative intensities of selected peaks, (2) principal component analysis (PCA).Clinical assessment of tumor radioresponse was carried out 4 months after first fraction of RT and the degree of the tumor shrinkage was determined as an index of radioresponsiveness (complete response (CR): 100% shrinkage, partial response (PR): ≥50% shrinkage, and no response (NR): ≤50% shrinkage) which was further correlated with the analysis of 2-RT serum chromatograms.ResultsNormal vs. malignant chromatograms demonstrated pronounced differences in the 800–1800 s region. Malignant vs. 2-RT chromatograms showed minute variations in the 1300–1800 s region. Our analysis, in both of the approaches, produced clear differentiation between ‘normal’ and ‘malignant’, whereas differentiation between ‘malignant’ and ‘2-RT’ was minimal. Clinical evaluation of the tumor radioresponse yielded that out of 13 patients (one patient discontinued the radiotherapy) ten showed CR, two showed PR and one NR. In case of prediction of tumor radioresponse, analysis of the 2-RT chromatograms produced only minor differentiation among CR, PR and NR groups.ConclusionProtein profiling of serum samples differentiated ‘normal’ from ‘malignant’, but could not differentiate ‘malignant’ from ‘2-RT’. Also this technique has limited application in prediction of tumor radioresponse.