Serotonin 1A receptor-mediated signaling through ERK and PKCα is essential for normal synaptogenesis in neonatal mouse hippocampus

Aberrant expression of the presynaptic serotonin 1A receptor (5-HT1A-R) because of a polymorphism in the 5-HT1A-R gene is associated with severe depression in human, whereas its absence up to postnatal day 21 (P21) in the forebrain of mice results in heightened anxiety in adulthood. These observations collectively indicate that the 5-HT1A-R has a crucial role in brain development. To understand the mechanistic underpinnings of this phenomenon, we used organotypic slice cultures of hippocampi from C57BL6 mice (C57) at P15, which coincides with the peak of neonatal synaptogenesis. Stimulation of the hippocampal 5-HT1A-R caused a dramatic increase in PSD95 expression and dendritic spine and synapse formation through sequential activation of the mitogen-activated protein kinase isozymes Erk1/2 and protein kinase C (PKC). Intrahippocampal infusion of 5-HT1A-R agonists and signaling inhibitors at P15 revealed that the same pathway through PKCα augments PSD95 expression and synaptogenesis in vivo in 24 h in both C57 as well as Swiss Webster mice. Furthermore, intrahippocampal infusion of the antidepressant fluoxetine, a serotonin reuptake inhibitor, also augmented PSD95 expression and synaptogenesis through the same pathway. This increased synaptogenesis was observed even 5 days after treatment. Finally, compared with the wild type, the 5-HT1A-R(−/−) mice harbor significantly less synapses in the hippocampus, but infusion of the PKC-stimulator and Alzheimer drug bryostatin into the 5-HT1A-R(−/−) mice to bypass the non-existent 5-HT1A-R boosted PSD95 expression and synaptogenesis. The elucidated signaling cascade explains how 5-HT1A-R regulates hippocampal sculpting and function, which may determine the affective phenotype of an adult

Molecular characterization of senescence marker protein-30 gene promoter: Identification of repressor elements and functional nuclear factor binding sites

<p>Abstract</p> <p>Background</p> <p>Senescence marker protein-30 (SMP30), whose expression declines during aging in rat liver, has been proposed as an important aging marker. Besides apoptosis, SMP30 also protects cells against various other injuries by enhancement of membrane calcium-pump activity. The mechanism of this differential gene expression mechanism is not known. DNA-protein interactions, mutation analysis and luciferase reporter assay studies have been performed to elucidate the mechanism of transcriptional regulation of SMP30 gene.</p> <p>Results</p> <p>We have characterized up to -2750 bp of the promoter by DNA-protein interactions studies. Twenty eight transcription factor binding sites have been identified by DNase I footprinting and electrophoretic mobility shift assay (EMSA). Transient transfection of 5' and 3' -deleted promoter-reporter constructs and luciferase assay illustrated the region between -128/+157 bp is sufficient to drive promoter activity. We have mapped an essential regulatory region between -513 to -352 bp which causes a drastic decline of reporter activity. This region contains CdxA, GATA2 and SRY transcription factor binding sites. Individual mutation of these three sites showed increase in reporter activity. Mutation in SRY site (-403/-368) showed maximum increase in reporter activity among these three sites. Therefore, we suggest that SRY like protein may be acting as a strong repressor of SMP30 gene along with CdxA and GATA-2. We also report that mutation of both Sp1 (172/-148 bp) and a C/EBPβ (-190/-177 bp) transcription binding site located adjacent to each other on SMP30 gene promoter, causes a significant enhancement in reporter activity than individual mutation, thus may be causing the repression of SMP30 promoter activity.</p> <p>Conclusion</p> <p>These studies provide novel insights into the mechanism that regulate SMP30 gene expression.</p

Corrosion behaviour of sintered ferrous alloys and Ferro-TiC in H₂SO₄and NaCl solutions

153-160The room temperature corrosion behaviour of plain carbon steel Fe-0.5C, low alloy steel Fe-5Cu-0.5C, tool steels ASP 23 and ASP 30, and ferro-TiC cermet was investigated in H2SO4 (0.1-5N) and 0.6 N NaCl solutions by potentiodynamic polarization technique. The study also includes X-ray diffraction, optical microscopy and scanning electron microscopy of the as-received as well as corroded surfaces. Porosities present in Fe-0.5C and Fe-5Cu-0.5C steels were found to be responsible for their poor corrosion resistance. ASP 30 steels were found to exhibit good corrosion resistance as compared to ASP 23 in both the media

Escherichia vulneris: an unusual cause of complicated diarrhoea and sepsis in an infant. A case report and review of literature

Escherichia vulneris is an opportunistic human pathogen. It has been primarily reported in adult patients and invasive infections have been observed in immune-suppressed individuals. This is the first report of E. vulneris causing complicated diarrhoea and sepsis in an infant. Two month old sick infant, born full-term, was admitted to the paediatrics department with loose motions and refusal to feed for four days. E. vulneris was isolated from blood in pure culture. The isolate was characterized for diarrhoeal virulence markers: heat labile and heat stable toxins (LT, ST) and hemolysin (hlyA) by PCR. The presence of LT enterotoxin and hemolysin provides strong evidence of the diarrhoeagenic potential of E. vulneris, further leading to the invasive infection triggering sepsis. As E. vulneris can lead to serious complications, an attempt should be made in clinical laboratories to identify and further characterize this new Escherichia species

Assessing the therapeutic usefulness of Ricinus communis: A multicentric observational clinical verification study

Introduction: Clinical verification is an ongoing research programme of the Central Council for Research in Homoeopathy, under which many symptoms of Indian and rarely used drugs in Homoeopathy have been clinically verified. Objectives: To clinically verify the symptomatology of Ricinus communis as observed during its proving conducted by Council and also to ascertain the clinical symptoms relieved in the process of verification. Materials and Methods: Two hundred and twenty-five patients from all age-groups and both sexes were enrolled from the outpatient departments (OPDs) of the institutes and units of the Council following the exclusion and inclusion criteria as per protocol and obtaining written consent. The presenting signs and symptoms were recorded in a predefined case recording proforma and if Ricinus communis was found very closely similar to the symptoms of the patient, the patients were enrolled in the study. The medicine was prescribed in different potencies as per the need of the case and in accordance with homoeopathic principles. The progress was noted in a follow-up sheet to determine the effects of the medicine, in relieving the symptoms of the patient. Result: Forty eight out of fifty three symptoms obtained from proving of Ricinus communis could be clinically verified. The characteristic indications were left-sided affinity, aggravation from sun, amelioration in open air, dryness of mucous membrane of gastrointestinal tract, dissatisfaction leading to irritability and anger. The usefulness of the medicine was mostly marked in relieving headache, coryza, aphthae, gastritis, diarrhoea, constipation and acne. All the verified symptoms indicated the scope of its therapeutic action. Conclusion: Ricinus communis can be considered as an important medicine for the management of acne, aphthae, backache, colic, constipation, coryza, cough, diarrhoea, dyspepsia, fever, gastritis, headache and irritability

A multicentric observational study to evaluate the role of homoeopathic therapy in vitiligo

Background: Vitiligo has an immense psychological impact on the affected individual and a reason for low self-esteem. Considering the disappointing outcomes, A multicentric open clinical study was undertaken by the Central Council for Research in Homoeopathy, at five institutes and units in India from October 2005 to September 2010. Aims and Objectives: This observational study aimed to see the usefulness of homoeopathic therapy in the management of vitiligo. Materials and Methods: 432 patients of all age groups suffering from vitiligo were enrolled in the study. Out of that, 169 patients completed 2 years of follow-up and were considered for analysis. Homoeopathic medicines, based on the totality of symptoms and repertorization were prescribed. The analysis of the cases was based on the Vitiligo Symptom Score (VSS) and photographs of the patients. Result was analyzed using statistical method of SPSS version 20. Results: The changes in the mean VSS at intervals of every 6 months was found to be statistically significant. Homoeopathic treatment was found to be useful in relieving vitiligo in varying degrees in 126 patients, out of which 4 (2.94%) cases showed marked improvement, 15 (11.03%) cases showed moderate improvement, 77 (56.62%) cases showed mild improvement, and 30 patients although improved, fell in the category of not significant improvement group (below 25% improvement). Ten homoeopathic medicines were found useful in the study of which Sulphur (n = 27), Arsenicum album (n = 19), Phosphorus (n = 19), and Lycopodium clavatum (n = 10) were the most commonly indicated and useful medicines

Coupling to a glioblastoma-directed antibody potentiates antitumor activity of curcumin

Current therapies for glioblastoma are largely palliative, involving surgical resection followed by chemotherapy and radiation therapy, which yield serious side effects and very rarely produce complete recovery. Curcumin, a food component, blocked brain tumor formation but failed to eliminate established brain tumors in vivo, probably because of its poor bioavailability. In the glioblastoma GL261 cells, it suppressed the tumor-promoting proteins NF-kappa B, P-Akt1, vascular endothelial growth factor, cyclin D1 and BClXL and triggered cell death. Expression of exogenous p50 and p65 subunits of NF-kappa B conferred partial protection on transfected GL261 cells against curcumin insult, indicating that NF-kappa B played a key role in protecting glioblastoma cells. To enhance delivery, we coupled curcumin to the glioblastoma-specific CD68 antibody in a releasable form. This resulted in a 120-fold increase in its efficacy to eliminate GL261 cells. A very similar dose response was also obtained with human glioblastoma lines T98G and U87MG. GL261-implanted mice receiving intratumor infusions of the curcumin-CD68 adduct followed by tail-vein injections of solubilized curcumin displayed a fourfold to fivefold reduction in brain tumor load, survived longer, and about 10% of them lived beyond 100 days. Hematoxylin-eosin staining of brain sections revealed a small scar tissue mass in the rescued mice, indicating adduct-mediated elimination of glioblastoma tumor. The tumor cells were strongly CD68+ and some cells in the tumor periphery were strongly positive for microglial Iba1, but weakly positive for CD68. This strategy of antibody targeting of curcumin to tumor comes with the promise of yielding a highly effective therapy for glioblastoma brain tumors