Factors Associated with Multidisciplinary Consultations in Patients with Early Stage Breast Cancer.

Purpose: Multidisciplinary care (MDC) encourages multiple specialists to formulate a unified treatment plan. We sought to determine the frequency and predictors of MDC and assess the association between MDC and nationally-recognized quality metrics in patients with breast cancer. Methods: We used the surveillance, epidemiology, and end results-medicare dataset to evaluate patients diagnosed with stages I-III breast cancer who underwent breast-conserving surgery between 2002 and 2011 with follow-up to 2012. We defined MDC as a visit claim from a surgeon, radiation oncologist and medical oncologist within 12 months of diagnosis. We used multivariable regression analysis to determine the association between demographic and clinical variables and MDC, and to assess the association between MDC and three nationally-recognized quality indicators (adjuvant hormone therapy for hormone receptor-positive tumors, chemotherapy for hormone receptor-negative cancer, and radiation after lumpectomy). Results: Of the 61,039 patients in our initial cohort, 53,849 (88.2%) saw a medical oncologist, 46,521 (76.2%) saw a radiation oncologist, and 43,280 (70.9%) were evaluated by all three providers the first year after diagnosis. MDC use was higher in patients with the highest socioeconomic status compared with the lowest [odds ratio (OR) 1.74, 95% CI 1.63-1.86], in patients diagnosed in later years, and those with stage III disease compared to stage I [OR 1.29, 95% CI 1.19-1.41]. Patients older in age (≥80 vs. 65-69 years, OR 0.33, 95% CI 0.31-0.34), patients with more comorbidities, those who lived in a rural setting compared to urban (OR 0.61, 95% CI 0.57-0.64), and unmarried patients (OR 0.79, 95% CI 0.76-0.82) were less likely to see all three providers. In a multivariable analysis, MDC use was associated with increased likelihood of meeting each quality metric. Conclusion: Early stage breast cancer patients were evaluated by a surgeon, radiation oncologist and medical oncologist less than 75% of the time. Enhanced coordination of care and navigation programs may improve the quality of care delivered.

Appendiceal neoplasms in Saudi Arabia: prevalence and clinicopathological profile.

BACKGROUND: The appendix is a small organ with no particular known function. Primary appendiceal neoplasms (ANs) are rare. While the prevalence is increasing worldwide over the past two decades, no apparent increase in the prevalence of ANs has been reported in the Arabian Gulf States. Recently, a significant decline in the age at diagnosis of some types of ANs has been reported worldwide, with a female predominance. OBJECTIVES: Evaluate the prevalence and clinicopathological characteristics of ANs within our institution in Saudi Arabia and compare them to limited existing studies from different regions as well as the Arabian Gulf States. DESIGN: Retrospective cohort. SETTING: Tertiary care center in Riyadh. PATIENTS AND METHODS: All patients who underwent appendectomy and had the appendix submitted for histopathological evaluation between May 2015 and June 2020 were included to allow for a follow-up of 5 years or more at the time of data collection. MAIN OUTCOME MEASURES: Demographics, clinical presentations, surgical interventions, histopathological findings, complications, and recurrence rates. SAMPLE SIZE: 25 AN patients. RESULTS: Of 1110 patients, 25 had ANs (13 female and 12 male participants) with a mean (standard deviation) age of 54.6 (14.1) years. Only 40% presented with acute appendicitis, 64% had comorbidities, and less than 50% underwent laparoscopic appendectomy. Histopathologically, 72% were low-grade appendiceal mucinous neoplasms (LAMNs). Complications were minimal grades (Clavien-Dindo classification), with 80% experiencing none. The mean hospital stay was 9.96 days. Local recurrence occurred in 8% of cases, and distant metastasis was documented in one adenocarcinoma case. However, the 5-year overall and disease-free survival rates were 88% and 80%, respectively. CONCLUSIONS: The incidence of ANs is increasing in Saudi Arabia with the higher prevalence of LAMNs. The pathological examination of the resected appendix played a pivotal role in the diagnosis of ANs. LIMITATIONS: Data collected retrospectively, a single institution, and a small population.

Bone Sialoprotein Is Critical for Alveolar Bone Healing in Mice.

Bone sialoprotein (BSP) is an extracellular matrix (ECM) protein associated with mineralized tissues, particularly bone and cementum. BSP includes functional domains implicated in collagen binding, hydroxyapatite nucleation, and cell signaling, although its function(s) in osteoblast and osteoclast differentiation and function remain incompletely understood. Genetic ablation of BSP in Ibsp knockout (Ibsp-/-) mice results in developmental bone mineralization and remodeling defects, with alveolar bone more severely affected than the femurs and tibias of the postcranial skeleton. The role of BSP in alveolar bone healing has not been studied. We hypothesized that BSP ablation would cause defective alveolar bone healing. We employed a maxillary first molar extraction socket healing model in 42-d postnatalIbsp-/- and wild-type (WT) control mice. Tissues were collected at 0, 7, 14, 21, and 56 d postprocedure (dpp) for analysis by micro-computed tomography (microCT), histology, in situ hybridization (ISH), immunohistochemistry (IHC), and quantitative polymerase chain reaction (qPCR) array. As expected, alveolar bone healing progressed in WT mice with increasing bone volume fraction (BV/TV), bone mineral density (BMD), and tissue mineral density (TMD), transitioning from woven to mature bone from 7 to 56 dpp. Ibsp messenger RNA (mRNA) and BSP protein were strongly expressed during alveolar bone healing in parallel with other osteogenic markers. Compared to WT, Ibsp-/- mice exhibited 50% to 70% reduced BV/TV and BMD at all time points, 7% reduced TMD at 21 dpp, abnormally increased Col1a1 and Alpl mRNA expression, and persistent presence of woven bone and increased bone marrow in healing sockets. qPCR revealed substantially dysregulated gene expression in alveolar bone of Ibsp-/- versus WT mice, with significantly disrupted expression of 45% of tested genes in functional groups, including markers for osteoblasts, osteoclasts, mineralization, ECM, cell signaling, and inflammation. We conclude that BSP is a critical and nonredundant factor for alveolar bone healing, and its absence disrupts multiple major pathways involved in appropriate healing.

Awareness and Acceptance of Digital Rectal Examination for the Clinical Evaluation of Anorectal Conditions Among the Saudi Population: A Cross-Sectional Study.

INTRODUCTION: Digital rectal examination (DRE) is an important diagnostic tool used by physicians to resolve several confusing clinical situations. The history and physical examination cannot be complete without performing a DRE. Any patient that presents with abdominal complaints (e.g., diarrhea, constipation, nausea, vomiting, abdominal or rectal pain, bleeding) needs a DRE which is important for detecting warning signs of serious conditions that require further investigation and evaluation such as malignancies. Therefore, our aim was to assess and measure the awareness of the Saudi population regarding the importance and acceptance to perform DRE. METHODS: This cross-sectional study was conducted in Riyadh, the capital city of Saudi Arabia, using an online survey between September 2022 and March 2023; the targeted participants were adults between the ages of 18 to 75. RESULTS: The study indicated that the general community awareness of DRE is low, with only 59.1% of participants having heard of DRE and 14.6% having undergone the procedure previously. The majority of individuals (60.9%) were willing to undergo DRE if a healthcare provider suggested it. Participants' knowledge of DRE's ability to detect various anorectal diseases varied. While the majority of individuals believed DRE could detect hemorrhoids, just 40.4% believed DRE could help detect colorectal cancer. Chronic constipation or diarrhea, feces-induced stretching, and prolonged sitting were the most oft-cited causes of hemorrhoids. Anemia was the most often reported consequence of hemorrhoids, followed by hypertension and diabetes. CONCLUSION: The significance of DRE as a screening tool for the early detection and prevention of anorectal problems, as well as the need for adequate care and treatment of hemorrhoids to prevent complications, are highlighted by these findings. Healthcare practitioners should actively recommend and provide information about DRE and other screening technologies, as well as address their patients' concerns and misconceptions.

Caecal Volvulus, Untwisting the Twisted: A Case Report and Literature Review.

Caecal volvulus is an uncommon surgical condition affecting mostly females in their second and third decade of life. It is of vital importance that the general surgeon recognises, resuscitates, diagnoses, and effectively treats these cases in a timely manner to maximise the chance of a positive outcome for the patient. Whilst there are several types of caecal volvulus, the treatment involves, in most cases, surgical intervention. There is a wide variety of surgical interventions that can be performed, ranging from caecopexy or fixation to lateral wall to performing a right hemicolectomy with primary ileocolic anastomosis. There are several factors that influence this decision and can also be based on an individual surgeon's expertise and experience. We present a case of a 21-year-old female who presented to our Emergency Department with lower abdominal pain, nausea, and vomiting. She was diagnosed with caecal volvulus with the aid of CT imaging, following which she underwent laparotomy in which caecal volvulus was noted. She underwent appendicectomy and caecopexy and was discharged after an uneventful recovery on post-operative day five and remains well on follow-up.

Prevalence and temporal trends of anemia in patients with heart failure.

BACKGROUND: Anemia is an important comorbidity in heart failure (HF), and it is associated with increased adverse disease experience and mortality. Previous reports have focused mainly on HF presenting in healthcare settings. We, therefore, set out to establish the nationwide prevalence and temporal trends of anemia in community-based patients with HF in the US. AIM: To establish the nationwide prevalence and temporal trends of anemia in community-based patients with HF in the US. DESIGN: The NHANES dataset, conducted by the CDC National Center for Health Statistics was used to collect nationally representative data on the health and nutritional status of the non-institutionalized US population. METHODS: We utilized the National Health and Nutrition Examination data collected over five survey cycles (2007-16). Included were participants aged 20-80 years with self-reported diagnosis of HF. Anemia was defined using 2 sex specific cut offs of 13 and 12 g/dl (cutoff 1), and 12 and 11 g/dl (cutoff 2), for men and women, respectively. The Chi square test was used to compare prevalence across different categories and survey cycles. Data analysis was done using STATA 16 with P-values < 0.05 considered statistically significant. RESULTS: The median hemoglobin in all HF patients was 13.5 g/dl (IQR 12.4-14.5). The prevalence of anemia among community-based patients with HF in the US was 21.34% (cutoff 1) and 9.03% (cutoff 2) and has been stable from 2007 to 2016. The burden of anemia was disproportionately higher in NH Blacks (34.48%, 95% CI 27.12-42.67) and those with BMI < 25 Kg/m2 (17.4%, 95% CI 10.9-26.64). CONCLUSION: The prevalence of anemia in patients with HF in the US is at least 9% and has remained stable over the past decade. This high persistent burden with limited proven interventions should spur further efforts aimed at identifying impactful ways of addressing anemia in patients with HF.

Dental defects in the primary dentition associated with hypophosphatasia from biallelic ALPL mutations.

ALPL encodes tissue-nonspecific alkaline phosphatase (TNAP), an enzyme expressed in bone, teeth, liver, and kidney. ALPL loss-of-function mutations cause hypophosphatasia (HPP), an inborn error-of-metabolism that produces skeletal and dental mineralization defects. Case reports describe widely varying dental phenotypes, making it unclear how HPP comparatively affects the three unique dental mineralized tissues: enamel, dentin, and cementum. We hypothesized that HPP affected all dental mineralized tissues and aimed to establish quantitative measurements of dental tissues in a subject with HPP. The female proband was diagnosed with HPP during childhood based on reduced alkaline phosphatase activity (ALP), mild rachitic skeletal effects, and premature primary tooth loss. The diagnosis was subsequently confirmed genetically by the presence of compound heterozygous ALPL mutations (exon 5: c.346G>A, p.A116T; exon 10: c.1077C>G, p.I359M). Dental defects in 8 prematurely exfoliated primary teeth were analyzed by high resolution micro-computed tomography (micro-CT) and histology. Similarities to the Alpl-/- mouse model of HPP were identified by additional analyses of murine dentoalveolar tissues. Primary teeth from the proband exhibited substantial remaining root structure compared to healthy control teeth. Enamel and dentin densities were not adversely affected in HPP vs. control teeth. However, analysis of discrete dentin regions revealed an approximate 10% reduction in the density of outer mantle dentin of HPP vs. control teeth. All 4 incisors and the molar lacked acellular cementum by micro-CT and histology, but surprisingly, 2 of 3 prematurely exfoliated canines exhibited apparently normal acellular cementum. Based on dentin findings in the proband's teeth, we examined dentoalveolar tissues in a mouse model of HPP, revealing that the delayed initiation of mineralization in the incisor mantle dentin was associated with a broader lack of circumpulpal dentin mineralization. This study describes a quantitative approach to measure effects of HPP on dental tissues. This approach has uncovered a previously unrecognized novel mantle dentin defect in HPP, as well as a surprising and variable cementum phenotype within the teeth from the same HPP subject.

Epidemiological study, immunohistochemistry and in situ hybridization studies of nasopharyngeal carcinomas: a Tunisian report.

Nasopharyngeal carcinomas (NPC) are a significant problem of public health in Tunisia. They are particular because of their characteristic geographic distribution. The aims of this study were, first, to appreciate the presence of Epstein-Barr virus (EBV) genome by immunohistochemistry (IHC) and in situ hybridization (ISH) and to compare their benefits to NPC diagnosis and, secondly, to verify the relation between NPC and factors bound to the food and environment conditions. Biopsies, recruited at the department of pathology of EPS Charles Nicolle at Tunis, were analyzed for EBV genome presence by ISH of EBV-encoded small RNA1 (EBER1). IHC was done with encoded nuclear antigen (EBNA1), latent membrane proteins (LMP1), and antigen BZ1 anti-Z EBV-replication activator (ZEBRA). An epidemiological study based upon the analysis of a detailed questionnaire submitted to patients (all from the north of Tunisia) and 60 witnesses was done. The statistic analysis was realised by SPSS Windows 11.5 Advanced Statistics. All samples were classified as Undifferentiated Carcinoma of Nasopharyngeal type (UCNT). We found a sex ratio of 2 with a bimodal repartition. ISH showed 96.6% positive samples. IHC revealed the EBV in 90% of cases and 66.7%, respectively, with EBNA1 and LMP1. The statistic analysis showed a meaningful relation (P<0.05, OR>3) between NPC and dietary factors (spices and piquant condiment), alcohol and the water quality.

Factors Associated With Follow-Up Care Among Women With Early-Stage Breast Cancer.

PURPOSE: Follow-up guidelines vary widely among national organizations for patients with early-stage breast cancer treated with curative intent. We sought to evaluate the patterns and predictors of provider follow-up care within the first 5 years after diagnosis. METHODS: Using the SEER-Medicare linked data set, we evaluated patients who were diagnosed with stage I and II breast cancer who underwent breast-conserving surgery from 2002 to 2007 with follow-up until 2012. We defined discontinuation of follow-up as > 12 months from the previous physician visit without a visit claim from either a surgeon, medical oncologist, or radiation oncologist. We performed a multivariable logistic regression and Cox proportional hazards regression analysis to determine factors associated with the discontinuation of follow-up care. RESULTS: Of the 30,053 patients enrolled in our initial cohort, 25,781 (85.8%) saw a medical oncologist and 21,612 (71.9%) saw a radiation oncologist in the first year in addition to a surgeon. Over the 5 years, 6,302 patients (21.0%) discontinued follow-up visits. Discontinuation of physician visits increased with increasing age. Women with stage II cancer ( v stage I) were less likely to discontinue follow-up visits (odds ratio, 0.78; 95% CI, 0.73 to 0.83). Time to early discontinuation was greater for patients with hormone receptor-negative tumors (hazard ratio, 1.14; 95% CI, 1.05 to 1.24). Women who were diagnosed more recently were less likely to discontinue seeing any physician. CONCLUSION: Twenty-one percent of patients with early-stage breast cancer discontinued seeing any oncology provider over the 5 years after diagnosis. Coordination of follow-up care between oncology specialists may reduce discontinuation rates and increase clinical efficiency.

High dietary cholecalciferol increases plasma 25-hydroxycholecalciferol concentration, but does not attenuate the hypertension of Dahl salt-sensitive rats fed a high salt diet.

The Dahl salt-sensitive rat, a model for salt-induced hypertension, develops hypovitaminosis D during high salt intake, which is caused by loss of protein-bound vitamin D metabolites into urine. We tested the hypothesis that high dietary cholecalciferol (5- and 10-fold standard) would increase plasma 25-hydroxycholecalciferol (25-OHD(3)) concentration (indicator of vitamin D status) of salt-sensitive rats during high salt intake. Salt-sensitive rats were fed 0.3% salt (low salt, LS), 3% salt (HS), 3% salt and 7.5 microg cholecalciferol/d (HS-D5), or 3% salt and 15 microg cholecalciferol/d (HS-D10) and sacrificed at week 4. Plasma 25-OHD(3) concentrations of the two groups of HS-D rats were similar to that of LS rats and more than twice that of HS rats. Urinary cholecalciferol metabolite content of HS-D rats was more than seven times that of HS rats. Systolic blood pressures of the hypertensive HS and HS-D rats did not significantly differ, whereas LS rats were not hypertensive. We conclude that high dietary cholecalciferol increases plasma 25-OHD(3) concentration, but does not attenuate the hypertension of salt-sensitive rats during high salt intake. Low salt intake may be necessary to both maintain optimal vitamin D status and prevent hypertension in salt-sensitive individuals.

Hyper immunoglobulin M immunodeficiency. (Dysgammaglobulinemia). Presence of immunoglobulin M-secreting plasmacytoid cells in peripheral blood and failure of immunoglobulin M-immunoglobulin G switch in B-cell differentiation.

The peripheral blood lymphocytes of nine patients with hyper immunoglobulin (Ig)M immunodeficiency were studied in an attempt to define the cellular basis of this disorder. B cells were normal in number but qualitatively abnormal in all patients. Approximately one-half of the B cell consisted of small lymphocytes (7-9 mum in diameter) bearing surface IgM and IgD, as well as C3 receptors. These cells were driven to secrete IgM but not IgG after in vitro stimulation by pokeweed mitogen. In the blood there were also large lymphocytes (10-14 mum in diameter) that possessed surface as well as intracytoplasmic IgM but lacked C3 receptors. These cells spontaneously secreted large amounts of IgM in vitro and on electron microscopy were found to be rich in rough endoplasmic reticulum. Such a subpopulation of lymphoid cells was not detected in normal peripheral blood and was unique for all patients with hyper IgM immunodeficiency studied.T cells from all patients were normal in number and in function both in vivo and in vitro and were able to generate adequate T-cell help to support IgG synthesis by normal B cells. No evidence was obtained for T cells capable of suppressing normal IgG synthesis in any of the patients after coculture with normal peripheral blood lymphocytes. The defect in hyper IgM immunodeficiency is intrinsic to B cells, which fail to switch from IgM to IgG synthesis.

Angiomyofibroblastoma of the spermatic cord: a case report.

Angiomyofibroblastoma (AMF) is a rare benign mesenchymal tumor with tendency to arise in the lower genital tract of middleaged women, predominately in the vulva. A few cases of AMF in males have been reported involving the scrotum, perineum or spermatic cord. We report a new case of AMF arising in the right inguinal region of a 27-year-old man. The tumor was well-circumscribed, myxoid and measured 30 mm in maximum dimension. On microscopic examination, the tumor was composed of spindle cells without atypia and with less than one mitosis figure per 10 high-power fields. Multinucleated cells and mast cells were observed. The stroma was myxoid and edematous with abundant capillary-sized blood vessels. Immunohistochemical staining showed a strong immunoreactivity for desmin and smooth muscle actin. The tumor cells were negative for estrogen receptors and focally positive for progesterone receptors with a low proliferative index of Ki67 (< 5%). This unusual neoplasm should be distinguished from aggressive angiomyxoma and other myxoid malignant tumors.

Associations Between Anthropometric Measurements and Cardiometabolic Risk Factors in White European and South Asian Adults in the United Kingdom.

OBJECTIVE: To investigate the association of 4 anthropometric measurements with cardiometabolic risk factors in a UK biethnic sample of South Asians (SAs) and white Europeans (WEs). PATIENTS AND METHODS: Baseline data were collected from adults of WE and SA origin participating in the Leicester arm of the Anglo-Danish-Dutch Study of Intensive Treatment in People with Screen Detected Diabetes in Primary Care (ADDITION-Leicester) study between August 2004 and December 2007. Overall, 6268 WE and SA adults had measures of body mass index, waist circumference, waist-to-hip ratio, and waist-to-height ratio assessed between June 18, 2004, and December 4, 2007. Hypertension, dyslipidemia, and dysglycemia were established from venous blood samples using standard definitions. Crude and adjusted (covariates used were age, sex, ethnicity, smoking, and alcohol consumption) odds ratios were calculated using multivariate logistic regression. Receiver operating characteristic curves and the area under the curve were used to calculate optimal cut points for the whole cohort and for both ethnic groups. RESULTS: Increases in all anthropometric measurements resulted in a higher odds ratio for each of the risk factors in both the crude and adjusted models (P<.001). The adjusted odds ratios for dyslipidemia, hypertension, and dysglygemia ranged from 1.30 to 1.35, from 1.36 to 1.52, and from 1.62 to 1.75 (P<.001 for all), respectively, in WEs. The adjusted odds ratio for dyslipidemia, hypertension, and dysglygemia ranged from 1.50 to 1.65 (P<.01), from 1.40 to 1.60 (P<.01), and from 1.96 to 2.11 (P<.001 for all), respectively, in SAs. The areas under the receiver operating characteristic curves for all the anthropometric measurements had low accuracy (P<.70) for the whole cohort and when stratified by ethnicity and sex. CONCLUSION: There is insufficient evidence to recommend replacing body mass index with another anthropometric measurement for the ethnically diverse population in the United Kingdom. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00318032.

[Endometrioid adenocarcinoma of the uterine cervix associated with mucinous ovarian cystadenocarcinoma]. / Association d'un adénocarcinome du col de type endométrioïde à un cystadénocarcinome mucineux de l'ovaire.

The authors report the case of a 40-year-old woman, who was operated for an ovarian mucinous cystadenocarcinoma. The pathologic findings of the hysterectomy specimen with bilateral salpingoophorectomy showed an ovarian mucinous cystadenocarcinoma associated with an endometrioid adenocarcinoma of the uterine cervix. The mucinous cystadenocarcinoma represents the third most common type of ovarian carcinoma. In the literature, this tumor had been found in association with endocervical adenocarcinoma or with minimal deviation adenocarcinoma (adenoma malignum) of the uterine cervix. However, its association with an endometrioid adenocarcinoma, to our knowledge, has not been reported.

Sleep deprivation and its associated factors among general ward patients at a tertiary care hospital in Pakistan.

OBJECTIVE: To estimate the occurrence rate of sleep deprivation and to identify the environmental, staff-related and patient-related factors associated with SD among general ward patients of a tertiary care hospital in Pakistan. METHODS: In a cross-sectional study, a pre-tested questionnaire was administered to 108 patients admitted into the general medical and general surgical wards of Aga Khan University Hospital, Karachi. RESULTS: In all, 50 (46.3%) respondents felt deprived of adequate sleep in the hospital. Worry about illness disturbed the night-time sleep of 47 (43.5%) patients; most of these had SD (70%) (p < 0.001). Other patients' noise disturbed 31.5% of study subjects and a significant majority (68%) of these had SD (p = 0.003). Over 17% of study subjects reported cell phone's ringing as a disturbing factor; more by those with SD (68%) compared to those with no SD (32%); again the difference was significant (p = 0.003). Physical discomfort and presence of cannula were reported as disturbing factors by 41.7% and 28.7% of the study subjects respectively but these were not significantly associated with SD. CONCLUSION: Our study revealed that sleep deprivation occurs commonly among general ward patients in tertiary care setting. Factors found to be associated with SD were amenable to modification to a greater extent.

[Low-grade central chondrosarcoma: difficult diagnosis in an adolescent girl]. / Chondrosarcome central de bas grade: un diagnostic difficile: à propos d'un cas.

Central chondrosarcoma of the tibia is exceptional, particularly in young patients. Low-grade tumors raise difficult problems for histological distinction with enchondroma. We report a case of grade 1 chondrosarcoma located in the upper portion of the tibia in a 17-year-old girl. After radical surgery, outcome was favorable with no recurrence or metastasis at three years follow-up. The distinction between low-grade central chondrosarcoma and enchondroma is one of the most difficult challenges in bone pathology. Clinical, radiographic and pathological data must be considered together to reach certain diagnosis.

Genetic determinants of hyperhomocysteinemia in atherosclerosis.

Hyperhomocysteinemia (Hhcy) is an independent risk factor for the development of atherosclerosis. The mechanisms by which HHcy promotes cardiovascular disease may be due to activation of pro-inflammatory factors, endoplasmic reticulum (ER) stress and oxidative stress. We aimed to study (i) gene mutations that cause HHcy. (ii) Estimation of inflammatory marker like ultrasenitive C-reactive proteins (hs-CRP) and total antioxidant levels (iii) determination of Hcy- dependent gene expression in vivo. 25 HHcy patients and 25 healthy controls were taken for this study. Mutation detection in MTHFR, CBS, MS and eNOS gene was by PCR-based restriction enzyme analysis and subsequently expression study was carried out by Reverse Transcriptase PCR and cloning technique. A significant association of HHcy with MTHFR (C677T) and MS (A2756G) genotype was observed (p<0.05). There was no association of Hhcy and eNOS genotype. The Hhcy patients, showed no expression of the ER stress gene, GRP78 in lymphocytes. Our study showed no effect of Hcy on the CD18 gene (pro-inflammatory pathway) expression, but a significant association of tHcy and hs-CRP levels in HHcy grp (t=2.28, p<0.05). This shows that HHcy induces inflammatory response, which could lead to tissue injury in the pathogenesis of the atherosclerotic process. Our findings show higher mRNA expression of manganese superoxide dismutase (Mn SOD) in HHcy group as compared to the control group. The Total Antioxidant Status (TAS) estimated was found to be significantly lower in the HHcy group as compared to healthy normals (t=4.8, p<0.01). Taken together these findings strongly suggest that the adverse effects of homocysteine are at least partly mediated by oxidative stress. Our study supports the hypothesis that Hcy evokes adverse vascular effects by promoting oxidative damage to endothelial cells.

A targeted DNA substrate mechanism for the inhibition of HIV-1 integrase by inhibitors with antiretroviral activity.

We recently reported that viral DNA could be the primary target of raltegravir (RAL), an efficient anti-HIV-1 drug, which acts by inhibiting integrase. To elucidate this mechanism, we conducted a comparative analysis of RAL and TB11, a diketoacid abandoned as an anti-HIV-1 drug for its weak efficiency and marked toxicity, and tested the effects of the catalytic cofactor Mg(2+) (5 mm) on drug-binding properties. We used circular dichroism and fluorescence to determine drug affinities for viral DNA long terminal repeats (LTRs) and peptides derived from the integrase active site and DNA retardation assays to assess drug intercalation into DNA base pairs. We found that RAL bound more tightly to LTR ends than did TB11 (a diketo acid bearing an azido group) and that Mg(2+) significantly increased the affinity of both RAL and TB11. We also observed a good relationship between drug binding with processed LTR and strand transfer inhibition. This unusual type of inhibition was caused by Mg(2+)-assisted binding of drugs to DNA substrate, rather than to enzyme. Notably, while RAL bound exclusively to the cleavable/cleaved site, TB11 further intercalated into DNA base pairs and interacted with the integrase-derived peptides. These unwanted binding sites explain the weaker bioavailability and higher toxicity of TB11 compared with the more effective RAL.