Anorectal Malformations and Associated Anomalies in Children.

Anorectal malformations (ARM) incorporate a broad spectrum of diseases, can affect both sexes, and involve the distal anus and rectum as well as the uro-genital tracts. Defects range from the minor which can be treated easily with an excellent outcome, to those are complex and often associated with other anomalies are difficult to manage with poor functional prognosis. This study was done to observe the hospital incidence of Anorectal malformations, frequency of types, sex distribution and spectrum of associations with ARM. The effects of presence of associated anomalies on morbidity and mortality also observed. Detailed history, clinical examinations and relevant investigations were performed for the primary and as well as the associated anomalies. A total of 80 patients were admitted in the department of pediatric surgery in Mymensingh Medical College Hospital during the period of June 2016 to May 2017. Age of the patients was ranging from 1-180 days with the mean age of 0.49±1.002 months. Male: Female ratio was 1.6: 1. Among them 48(60%) were high and 32(40%) were low variety of ARM. In male 37(46.2%) had high and 13(13.7%) were low variety whereas in female 11(13.7%) were high and 19(23.7%) had low ARM. Associated anomalies were seen in 25(31.2%) patient -18 in males and 7 in females; 20 in high and 5 in low ARM. Associated anomalies were uro-genital 11(13.8%), cardiovascular 10(12.5%), vertebral 4(5%), limb defects 3(3.5%) and others 2(2.5%). Four patients have more than one anomaly. Anorectal malformations occur more in boys than girls. Males were more likely to have high lesions and without fistula was the common defect. Low variety ARM were found more in females with Anovestibular fistula is the commonest defect. The most common associated anomalies were recto urinary fistula (13.8%). Associations were more in high than low ARM but not significant (p>0.05). Post operative complications were more in high ARM in both sexes with associated anomalies. The effects of types and associations on morbidity and mortality were significantly different (p<0.05).

The Buffalo study: Outcome and associated predictors in endodontic microsurgery- a cohort study.

AIM: To (i) evaluate and compare the outcome of endodontic microsurgery (EMS) using periapical radiographs (PAs) and cone-beam computed tomography (CBCT) scans; (ii) identify prognostic factors affecting the outcome; and (iii) correlate the effect of guided tissue regeneration (GTR) on the pattern of apical bone remodelling. METHODOLOGY: Eighty-two patients (101 teeth) who received EMS were included and followed-up using clinical and radiographic examinations (PAs and CBCT scans). Two calibrated endodontists evaluated the radiographic healing (favourable or unfavourable) by assessing PAs and CBCT. The success (favourable radiographic outcome with no clinical symptoms) and survival rates (tooth retention without clinical symptoms) were calculated, and the cause of failure (diseased or fractured) was identified. Pre-treatment (age, sex, tooth type, position, sequence of treatment, quality of root canal before surgery, presence/absence of through-and-through lesion, presence/absence apico-marginal defect) and treatment (presence/absence of errors during surgery, type of error (major or minor), retro-preparation depth, presence/absence of an isthmus, retro-filling material used, presence/absence bone graft material and/or resorbable membrane) factors were recorded. Data were analysed statistically to determine the inter-observer, intra-observer and inter-radiographic agreements. Univariate, bivariate and logistic regression analysis were used to determine prognostic factors affecting the outcome and the effect of GTR on the pattern of apical bone remodelling. The significance level was set at 5%. RESULTS: Sixty-eight patients (83 teeth) presented for outcome evaluation (recall rate: 84%). The survival rate was 93%. The success rate was 88% using PA and 86% using CBCT when vertical root fracture (VRF) cases were included and 94% using PAs, and 91% using CBCT when VRF cases were excluded. The intra- and inter-observer agreements were substantial using CBCT, slight to a fair agreement using PA (P < 0.001), and slight to moderate for inter-radiographic agreement. The occurence of a major procedural error during surgery was the only negative predictor for the outcome of EMS (P = 0.013). GTR did not affect the success rate or the type of healing when assessed using PA but it affected the type of healing on CBCT scans (complete vs incomplete healing) and the pattern of cortical plate remodelling (P < 0.001). CONCLUSION: The success and survival rate of endodontic microsurgery was very high, and the occurrence of a major procedural error during surgery was the only factor affecting the outcome. GTR did not improve the outcome, but did affect the quality of apical bone remodelling following EMS.

Prevalence of Intestinal Parasites in Raw Vegetables Consumed by Inhabitants of Mymensingh City.

Uncooked vegetables are an important part of a healthy diet in different parts of the world. The present descriptive cross sectional type of study was carried out to determine the parasitological contamination of vegetables sold at markets of Mymensingh city of Bangladesh. Parasitological examinations were performed in material derived from 200 specimens of 10 different vegetables to detect intestinal parasites in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from July 2015 to July 2016. Each sample was washed with 5 liter distilled water, then washing fluid was centrifuged and resulting sediment was collected for iodine and normal saline wet mount slide preparation to detect human pathogenic intestinal parasites by microscopic examination. Out of 200 vegetable samples, 123(61.5%) were contaminated with different species of parasites. The vegetables red amaranth had the highest contamination rate of 18/20 (90%) followed by jute leaf 17/20 (85%), coriander leaf 15/20 (75%), onion 14/20 (70%), lady's finger 14/20 (70%), radish 13/20 (65%), green pepper 12/20 (60%), carrot 12/20 (60%), cucumber 5/20 (25%), tomato 3/20 (15%). Among the contaminating parasites Ascaris lumbricoides (egg) 36.5% was the most prevalent followed by Strongyloides stercoralis (larva) 35.5%, Entamoeba histolytica ( trophozoite and different cystic stages) 8.5%, Hook worm (egg and larva) 6.5% , Balantidium coli (trophozoite) 4.0%, Enterobius vermicularis (egg) 3.5%, Trichuris trichiura (egg) 1.5%, Giardia lamblia (cyst) 1.0%. To our knowledge, it was the first base line study pursued in Bangladesh. Contamination of raw vegetables from markets with pathogenic parasites would increase the risk of disease to the population that consumes or works with these products. So improperly washed raw eaten vegetables should be considered a potential risk for contracting parasites, particularly helminthes in Mymensingh city.

PO-42 - A novel approach to a common problem: after-hours care of cancer patients diagnosed with VTE.

INTRODUCTION: Oncologists often receive phone calls from radiologists after regular working hours or while on call, informing them that a cancer patient has been diagnosed with a blood clot. In these situations, there may not be nursing staff available to contact the patient and provide teaching for Low Molecular Weight Heparin (LMWH) injections. As a result, patients are often sent to the emergency for injection and teaching, taxing an already overburdened emergency department. This problem constitutes an important care gap. AIM: In Alberta, Canada, pharmacists are able to prescribe medications including LMWH. We designed an after-hours program to provide care for cancer patients diagnosed with VTE. MATERIALS AND METHODS: Once the oncologist is made aware of the patient with a clot, a simple one page document is filled out and faxed to a 24-hour outpatient pharmacy outlining the following: patient demographics, clot location, systemic therapy, current anticoagulant and anti-platelet agents. The oncologist has the option to specify desired LMWH. The patient goes to the pharmacy where the pharmacist weighs the patient, reviews blood work electronically and prescribes the LMWH. Also provided are injection teaching and telephone follow-up. A specific algorithm is followed with the pharmacist able to contact the on call oncologist in specific situations where the patient's condition falls outside of the algorithm guideline. The pharmacist is able to order blood work, particularly to evaluate for Heparin Induced Thrombocytopenia. Patients must follow up with their oncologist within 7 days of diagnosis. RESULTS: This program has been run as a pilot and preliminary data will be presented at the ICTHIC meeting. Specifically, we will assess usage of the program, appropriateness of therapy chosen according to Canadian practice guidelines, as well as patient, pharmacist and physician satisfaction with the program. CONCLUSIONS: We believe that this outpatient pharmacy program is innovative, will decrease burden on emergency departments, and takes advantage of our pharmacists' ability to independently assess patients and write prescriptions. This program may serve as a model for other cancer centers looking for a novel way to provide after-hours care of patients diagnosed with VTE.

Vitamin E attenuates neurotoxicity induced by deltamethrin in rats.

BACKGROUND: The safety of Deltamethrin (DM) has been raised as a point of concern. The current investigation was envisaged to explore the responsiveness of oxidative stress parameters, DNA fragmentation and expression levels of TP53, cycloxygenase 2 (COX2) and cytochrome p4502E1 (CYP2E1) as toxicological endpoint in rats treated with DM. as well as attention was provided to the neuroprotective effect of vitamin E (VE). METHODS: Four different groups of rats were used in this study, group I served as control, group II received DM (0.6 mg/kg BW), group III received both DM plus VE and finally group IV received VE only (200 mg/kg BW). The treatment regimen was extending for one month for all groups and the brain tissues were collected for further analysis. RESULTS: The obtained results showed a highly statistically significant increase in lipid peroxidation (LPO) content, nitric oxide concentration, and DNA fragmentation percentage and expression level of CYP2E1, TP53 and COX2 genes, in addition statistical significant reduction in total antioxidant capacity in DM treated group as compared to control were detected. Oral administration of VE attenuated the neurotoxic effects of DM through improvement of oxidative status, DNA fragmentation percentage and suppressing the expression level of CYP2E1, TP53 and COX2 genes. CONCLUSION: From this study we concluded that VE supplementation has beneficial impacts on DM neurotoxicity in rats through its antioxidant and antiapoptotic properties.

Prognostic significance of ploidy and S-phase fraction in primary intraoral squamous cell carcinoma and their corresponding metastatic lymph nodes.

BACKGROUND: Despite improvements in diagnosis and therapy of oral and oro-pharyngeal carcinomas during the past 30years the 5-year disease-free survival is still poor. Patient's prognosis is affected by cervical lymph node metastasis rather than primary tumors. The DNA ploidy and S-phase fraction (SPF) are associated with tumor aggressiveness and patient outcome in many solid tumors. PURPOSE: Analysis of DNA ploidy and SPF in primary oral squamous cell carcinoma (OSCC) and corresponding node metastasis as prognostic markers in relation to conventional prognostic factors and disease-free survival (DFS). METHODS: Ploidy status and SPF (mean value) of 37 formalin-fixed paraffin embedded (FFPE) primary OSCC tumors and their corresponding lymph node metastasis were assessed by flow cytometry (FCM) and correlated with clinicopathologic prognostic parameters and DFS. RESULTS: Most of OSCC tumors (86.5%) were Grade II. Among primary OSCC the incidence of aneuploidy was 19%, 51.4% showed high SPF (>10.62%) and 48.6% had low SPF (<10.62%). Border line significance (P=0.10) was detected between ploidy status and SPF in primary tumors. In lymph node metastases all tumors were diploid, 78.4% of metastatic tumors revealed low SPF and only 21.6% showed high SPF. There was a statistically significant correlation (p=0.02) between site of tumors and DFS and a highly statistically significant correlation (p=0.01) between SPF of primary tumors and DFS. CONCLUSIONS: High SPF of primary OSCC tumors assessed by FCM was significantly associated with decreased disease free survival rates. DNA ploidy showed no relationship to bad prognostic indicators in either primary OSCC or their metastatic tumors.

Memantine prevents aluminum-induced cognitive deficit in rats.

Memantine, a noncompetitive NMDA receptor blocker, has been demonstrated to be neuroprotective against various neurotoxins. Aluminum, a well-known neurotoxin, has been suggested to be a contributing factor in Alzheimer's disease. In this study we investigated the possible effect of memantine on aluminum-induced cognitive impairment in rats. Rats were exposed to aluminum chloride (100mg/kg/day) and memantine (5, 10 and 20mg/kg/day) for 60 days. Cognitive functions were evaluated using three tests: Morris water maze, radial arm maze and passive avoidance tests. Results showed that memantine failed at low doses to have any significant influence on aluminum-induced memory deficit, but the 20mg/kg dose was found to cause significant enhancement of memory in the aluminum-exposed rats. This is the first study to demonstrate the protective role of memantine against aluminum-induced neuronal dysfunction. Biochemical and histological investigations are highly indicated to clarify the possible pharmacodynamic basis.

Rivastigmine reverses aluminum-induced behavioral changes in rats.

Aluminum, a known neurotoxin, has long been implicated in the pathogenesis of Alzheimer's disease. Its exposure is associated with impairment in the cholinergic system in the brain. In this study we investigated the behavioral effects of aluminum in rats and the possible effect of rivastigmine, a cholinesterase inhibitor, on the aluminum-induced behavioral changes. Rats were exposed to aluminum chloride (100 mg/kg/day i.p.) for 60 days before the start of behavioral tests. Rivastigmine was given in doses of 0.5, 1, 1.5 and 2.5 mg/kg i.p. 60 min before the behavioral tests. Five tests were investigated; open field test, Morris water maze, radial arm maze, passive avoidance test and rota-rod test. Results showed that aluminum exposure was associated with significant reductions in spontaneous locomotor and exploratory activities in open field test and significant impairments in learning and memory in Morris water maze, radial arm maze and passive avoidance tests. The behavioral impairments caused by aluminum were significantly improved by rivastigmine. Neither aluminum alone nor co-treatment with rivastigmine caused any significant alteration of the animals' performance in rota-rod test. The improvements in activity, learning and memory caused by rivastigmine were found to be dose-dependent, and the maximal improvement was encountered with its large dose (2.5 mg/kg). From these results we can conclude that rivastigmine can reverse behavioral deficits caused by aluminum intoxication.

Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers

Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13 percent). It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis.

Functional genomics of silencing TREM-1 on TLR4 signaling in macrophages.

Triggering receptor expressed on myeloid cells 1 (TREM-1) is a recently discovered molecule that is expressed on the cell surface of monocytes and neutrophils. Engagement of TREM-1 triggers synthesis of proinflammatory cytokines in response to microbes, but the extent and mechanism by which TREM-1 modulates the inflammatory response is poorly defined. In the present study, we investigated the functional effects of blocking TREM-1 on the Toll-like receptor (TLR)4-mediated signaling pathway in macrophages. By transfecting cells with small hairpin interfering RNA molecules to TREM-1 (shRNA), we confirmed that TREM-1 mRNA and protein expression was greatly attenuated in RAW cells in response to treatment with LPS. PCR array for genes related to or activated by the TLR pathway revealed that although the expression of TLR4 itself was not significantly altered by silencing of TREM-1, expression of several genes, including MyD88, CD14, IkappaBalpha, IL-1beta, MCP-1, and IL-10 was significantly attenuated in the TREM-1 knockdown cells in response to treatment with LPS. These data indicate that expression of TREM-1 modulates the TLR signaling in macrophages by altering the expression of both adaptor and effector proteins that are critical to the endotoxin response.

Biodegradable microparticulates of beta-estradiol: preparation and in vitro characterization.

Beta-estradiol has been recommended for the long-term therapy of osteoporosis and its oral formulations are subjected to intensive first pass metabolism. The present investigation was aimed at preparing and characterizing biodegradable microparticles of beta-estradiol with polymers such as PLA, PLGA 85/15, PLGA 75/25, and their mixtures. The microparticles were prepared by solvent evaporation method using methylene chloride as a solvent and polyvinyl alcohol as a surfactant. The drug-polymer ratios were 1:3, 1:5, and 1:7. The prepared microparticles (twelve formulations) were tested for encapsulation efficiency and in vitro drug release in 50% methyl alcohol/phosphate buffer pH 7.4. The results showed that the encapsulation efficiency varied from 81 to 100% and the formulation fabricated from PLGA 85/15 (1:3) showed less burst and consistent long time release. This formulation when further characterized displayed irregular spherical shape with an average particle size of 72 microm. The crystallinity of the drug was reduced when investigated using X-ray diffractometry. No chemical interaction between the drug and the polymer was observed as evidenced by FT-IR analysis. The results indicated that beta-estradiol biodegradable microparticles with PLGA 85/15 (1:3) could be a suitable approach for long term therapy of osteoporosis.

Preliminary investigation into the effects of electrical stimulation on mandibular distraction osteogenesis in goats.

Craniofacial distraction osteogenesis (DO) was found to be a procedure with low preoperative and postoperative morbidity. Direct current electrical stimulation is used clinically to treat different orthopaedic problems. It provided a significant increase in new-bone formation, and a higher mechanical strength of healing. The purpose of this study was to test the effect of electric current on distraction osteogenesis and to establish the best period to apply an electric current during the different distraction phases. Twelve healthy adult goats were subjected to a vertical osteotomy in the symphyseal area. A distraction device was fixed to the bone. Animals were divided into two groups, Group I (three goats) and group II (nine goats). In group I, the device was activated 1mm per day for 10 days; while in group II, the device was activated 1mm per day for 10 days with the application of direct electric current stimulation of 10 microA either during the first 3 days of latency in a continuous mode (ESL group), or during the first 3 days of the activation period in a continuous mode (ESA group); or during the first 3 days of the consolidation period in a continuous mode (ESC group). After the activation period was completed, the distraction device left in place for additional 15 days for bone consolidation in all animals. Animals were then sacrificed, the mandible was harvested, and the distracted areas were removed and processed for mechanical and histological studies. The results of this study suggested that direct current electrical stimulation display synergism on mandibular distraction when this stimulation applied to the distraction zone during activation or consolidation periods.

Novel mutations in spastin gene and absence of correlation with age at onset of symptoms.

Autosomal dominant hereditary spastic paraplegia is genetically heterogeneous, with at least five loci identified by linkage analysis. Recently, mutations in spastin were identified in SPG4, the most common locus for dominant hereditary spastic paraplegia that was previously mapped to chromosome 2p22. We identified five novel mutations in the spastin gene in five families with SPG4 mutations from North America and Tunisia and showed the absence of correlation between the predicted mutant spastin protein and age at onset of symptoms.

Loss of heterozygosity on 8p in prostate cancer implicates a role for dematin in tumor progression.

Dematin is a cytoskeletal protein that bundles actin filaments in a phosphorylation-dependent manner. The primary structure of dematin is organized into an N-terminal core domain of unknown function and a C-terminal domain that is homologous to the "headpiece" domain of villin. We have previously localized the dematin gene on human chromosome 8p21.1, a region distal to the ankyrin locus for hereditary spherocytosis. Radiation hybrid mapping now places dematin between D8S258 and D8S137, two microsatellite markers frequently deleted in prostate cancer. The 8p21.1 region is also deleted in prostate, breast, colon, and bladder cancers, suggesting the presence of a tumor suppressor gene(s). Using laser-capture microdissection technique and fluorescence in situ hybridization (FISH), we demonstrate loss of heterozygosity (LOH) of the dematin gene in a majority of chromosomal region 8p21-linked prostate tumors. One allele of dematin was also deleted in the established prostate adenocarcinoma cell line PC-3, which displays a classic oncogenic phenotype. Overexpression of wild-type dematin in PC-3 cells resulted in the restoration of a more polarized, epithelial-like phenotype. Conversely, the heterologous expression of dominant negative mutants of dematin perturbed normal cell morphology of NIH 3T3 fibroblasts. These results suggest a biological function of dematin in the regulation of cell shape, with implications in the pathobiology of prostate tumorigenesis.

Alternative splicing and structure of the human erythroid dematin gene.

Human erythroid dematin is a cytoskeletal protein capable of bundling actin filaments in vitro. The carboxyl terminal domain of dematin is homologous to the headpiece domain of villin, an actin-binding protein of the brush border cytoskeleton. Here we report the complete structure of the dematin gene located on human chromosome 8p21.1, a region frequently deleted in prostate cancer. The dematin gene is composed of 15 exons spanning approximately 15 kb. We also report two novel isoforms of dematin derived from alternative splicing of the dematin gene in the brain.

Lipase-catalysed selective deacetylation of phenolic/enolic acetoxy groups in peracetylated benzyl phenyl ketones.

Highly chemo- and regioselective de-esterification has been observed in the deacetylation of peracetylated enolic forms of polyphenolic benzyl phenyl ketones by lipase from porcine pancreas (PPL) suspended in tetrahydrofuran (THF). The enzyme selectively deacetylates the enolic acetoxy over the phenolic acetoxy group(s) and continuation of the reaction resulted, in addition the regioselective deacetylation of acetoxy function para to the nuclear carbonyl group.

Human erythrocyte dematin and protein 4.2 (pallidin) are ATP binding proteins.

Dematin and protein 4.2 are peripheral membrane proteins associated with the cytoplasmic surface of the human erythrocyte plasma membrane. Isoforms of dematin and protein 4.2 exist in many nonerythroid cells. In solution, dematin is a trimeric protein containing two subunits of 48 kDa and one subunit of 52 kDa. Recent determination of the primary structure of the 52 kDa subunit of dematin showed that it contains an additional 22-amino acid sequence in the headpiece domain. An alignment of the 22-amino acid insertion sequence revealed that the 52 kDa subunit of dematin shares a novel 11-amino acid motif with protein 4.2. In this communication, we report that the conserved 11-amino acid motif in dematin52 and protein 4.2 contains a nucleotide binding P-loop. Direct binding of ATP is demonstrated to the glutathione S-transferase fusion proteins containing corresponding segments of dematin52 and protein 4.2 as well as to purified protein 4.2. The binding of ATP to the recombinant domains of dematin52 and protein 4.2 is specific, saturable, and of high affinity. The nucleotide specificity of the P-loop is restricted to ATP since no detectable binding was observed with GTP. These results show that the 11-amino acid motif provides an ATP binding site in dematin52 and protein 4.2. Although the functional significance of ATP binding is not yet clear, our findings open new perspectives for the function of dematin and protein 4.2 in vivo.

Clinical down staging of cancer cervix with cytology.

A prospective study was carried out on 1500 patients attending the Gynaecology outpatient department of three referral hospitals in Dhaka City. These patients were selected on some selected clinical criteria to correlate several clinical markers (clinical down staging) with that of Pap's smear for early detection and secondary prevention of cancer cervix. Speculum examination and Pap's smear were done in all cases. Among 1500 women, 224 had abnormal Pap's smear. Histologic examination of specimens from these cases showed chronic cervicitis in 45 cases, dysplasia in 84 and squamous cell carcinoma in 95. Carcinoma was found in 130. Thus, there were 297 Pap's smear negative cases were biopsied on clinical ground. 225 cases of invasive carcinoma. From this study, it seems that clinical downstaging with cytology programme has to be started immediately throughout the country for diagnosis of carcinoma of the cervix in its preinvasive and potentially curable stage.

DLG1: chromosome location of the closest human homologue of the Drosophila discs large tumor suppressor gene.

The Drosophila discs large tumor suppressor protein, Dlg, is the prototype of a newly discovered family of proteins termed MAGUKs (membrane-associated guanylate kinase homologues). MAGUKs are localized at the membrane-cytoskeleton interface, usually at cell-cell junctions, where they appear to have both structural and signaling roles. They contain several distinct domains, including a modified guanylate kinase domain, an SH3 motif, and one or three copies of the DHR (GLGF/PDZ) domain. Recessive lethal mutations in the discs large tumor suppressor gene interfere with the formation of septate junctions (thought to be the arthropod equivalent of tight junctions) between epithelial cells, and they cause neoplastic overgrowth of imaginal discs, suggesting a role for cell junctions in proliferation control. A homologue of the Dlg protein, named Hdlg, has been isolated from human B lymphocytes. It shows 65-79% identity to Dlg in the different domains, and it binds to the cytoskeletal protein 4.1. Here, we report that the gene for lymphocyte Hdlg, named DLG1, is located at chromosome band 3q29. This finding identifies a novel site for a candidate tumor suppressor on chromosome 3.