Novel IRF6 Mutations Detected in Orofacial Cleft Patients by Targeted Massively Parallel Sequencing.

Common variants in interferon regulatory factor 6 ( IRF6) have been associated with nonsyndromic cleft lip with or without cleft palate (NSCL/P) as well as with tooth agenesis (TA). These variants contribute a small risk towards the 2 congenital conditions and explain only a small percentage of heritability. On the other hand, many IRF6 mutations are known to be a monogenic cause of disease for syndromic orofacial clefting (OFC). We hypothesize that IRF6 mutations in some rare instances could also cause nonsyndromic OFC. To find novel rare variants in IRF6 responsible for nonsyndromic OFC and TA, we performed targeted multiplex sequencing using molecular inversion probes (MIPs) in 1,072 OFC patients, 67 TA patients, and 706 controls. We identified 3 potentially pathogenic de novo mutations in OFC patients. In addition, 3 rare missense variants were identified, for which pathogenicity could not unequivocally be shown, as all variants were either inherited from an unaffected parent or the parental DNA was not available. Retrospective investigation of the patients with these variants revealed the presence of lip pits in one of the patients with a de novo mutation suggesting a Van der Woude syndrome (VWS) phenotype, whereas, in other patients, no lip pits were identified.

Tooth agenesis and orofacial clefting: genetic brothers in arms?

Tooth agenesis and orofacial clefts represent the most common developmental anomalies and their co-occurrence is often reported in patients as well in animal models. The aim of the present systematic review is to thoroughly investigate the current literature (PubMed, EMBASE) to identify the genes and genomic loci contributing to syndromic or non-syndromic co-occurrence of tooth agenesis and orofacial clefts, to gain insight into the molecular mechanisms underlying their dual involvement in the development of teeth and facial primordia. Altogether, 84 articles including phenotype and genotype description provided 9 genomic loci and 26 gene candidates underlying the co-occurrence of the two congenital defects: MSX1, PAX9, IRF6, TP63, KMT2D, KDM6A, SATB2, TBX22, TGFα, TGFß3, TGFßR1, TGFßR2, FGF8, FGFR1, KISS1R, WNT3, WNT5A, CDH1, CHD7, AXIN2, TWIST1, BCOR, OFD1, PTCH1, PITX2, and PVRL1. The molecular pathways, cellular functions, tissue-specific expression and disease association were investigated using publicly accessible databases (EntrezGene, UniProt, OMIM). The Gene Ontology terms of the biological processes mediated by the candidate genes were used to cluster them using the GOTermMapper (Lewis-Sigler Institute, Princeton University), speculating on six super-clusters: (a) anatomical development, (b) cell division, growth and motility, (c) cell metabolism and catabolism, (d) cell transport, (e) cell structure organization and (f) organ/system-specific processes. This review aims to increase the knowledge on the mechanisms underlying the co-occurrence of tooth agenesis and orofacial clefts, to pave the way for improving targeted (prenatal) molecular diagnosis and finally to reflect on therapeutic or ultimately preventive strategies for these disabling conditions in the future.

Anticancer Effects of Extracts from the Fruit of Morinda Citrifolia (Noni) in Breast Cancer Cell Lines.

Morinda citrifolia L. (NONI) fruits have been used for thousands of years for the treatment of many health problems including cancer, cold, diabetes, flu, hypertension, and pain. Plant extracts have reported several therapeutic benefits, but extraction of individual compound from the extract often exhibits limited clinical utility as the synergistic effect of various natural ingredients gets lost. They generally constitute polyphenols and flavonoids. Studies have suggested that these phytochemicals, especially polyphenols, display high antioxidant properties, which help to reduce the risk of degenerative diseases, such as cancer and cardiovascular diseases. Several in-vitro and in-vivo studies have shown that Noni fruits have antioxidant, anti-inflammatory, anti-dementia, liver-protective, anticancer, analgesic, and immunomodulatory effects. Till date about 7 in vitro cancer studies have been done, but a detailed in vitro study including cell cycle and caspase activation assay on breast cancer cell line has not been done. In the present study different Noni fruit fractions have tested on cancer cell lines MCF-7, MDA-MB-231 (breast adenocarcinoma) and one non-cancer cell line HEK-293 (Human embryonic kidney). Out of which ethylacetate extract showed a higher order of in vitro anticancer activity profile. The ethylacetate extract strongly inhibited the proliferation of MCF-7, MDA-MB-231 and HEK-293 cell lines with IC50 values of 25, 35, 60 µg/ml respectively. The extract showed increase in apoptotic cells in MCF-7 and MDA-MB-231 cells and arrested the cell cycle in the G1/S phase in MCF-7 and G0/G1 phase in MDA-MB-231 cells. Noni extract also decreases the intracellular ROS generation and mitochondrial membrane potential.

Long-term efficacy of single-dose radiofrequency-induced heat therapy vs. intralesional antimonials for cutaneous leishmaniasis in India.

BACKGROUND: Radiofrequency-induced heat therapy (RFHT) has been found to be safe and effective against cutaneous leishmaniasis (CL) in the short term, but its long-term efficacy is unclear. OBJECTIVES: To compare the long-term efficacy of RFHT vs. intralesional sodium stibogluconate (SSG) injections in the treatment of CL in India. METHODS: One hundred patients with a confirmed diagnosis of CL were randomly assigned in a 1 : 1 ratio to receive topical RFHT for 30-60 s or seven intralesional injections of SSG (50 mg cm(-2) of lesion). Improvement and recurrence were monitored every 15 days after the initiation of treatment for 4 months and then at 5, 6, 9, 12 and 18 months post-treatment; the rates of complete cure were compared. RESULTS: Lesions were healed in 47 out of 50 patients (94%) in the RFHT group and in 46 out of 50 patients (92%) in the SSG group at week 12. Time to complete healing was comparable in the two groups. At 6 months post-treatment, cure rates in the RFHT and SSG groups were 98% [95% confidence interval (CI) 94-100%] and 94% (95% CI 86-100%), respectively. Age, sex and lesion size or number had no effect on cure rates. No relapse of infection was recorded in cured patients in either group up to 12-18 months after initiation of treatment. Skin biopsies of cured lesions in eight out of eight (100%) patients from the RFHT group and three of three from the SSG group at 12 months showed minimal fibrosis and were negative for Leishmania tropica by polymerase chain reaction test. CONCLUSIONS: A single application of RFHT is safe, cosmetically acceptable and effective in inducing a long-term cure of CL.

Ruptured Aortic Aneurysm Secondary to Psoas abscess after Intravesical Bacilli Calmette-Guérin.

Bacillus Calmette-Guérin (BCG) is a live attenuated strain of Mycobacterium bovis that has been used as effective treatment for early-stage transitional carcinoma of the urinary bladder. We present a case of a 68 year old man who had an abdominal aortic aneurysm following BCG therapy for bladder cancer. Contrast enhanced computerized tomogram (CECT) of abdomen and pelvis revealed bilateral hypodense lesions suggestive of psoas abscesses. In addition, a saccular abdominal aortic aneurysm measuring 4x3.6 cm involving infrarenal aorta with surrounding hematoma was seen. At surgery, he was found to have a psoas abscess and hemorrhage. He underwent ligation of the aorta and an axillary-bifemoral bypass. He was given one year of anti-tubercular therapy to which he responded clinically.

Population-based stroke survey in Mumbai, India: incidence and 28-day case fatality.

OBJECTIVES: The aims of this study were (1) to establish a prospective community-based stroke registry in Mumbai of subjects having 'first-ever stroke' (FES) and (2) to collect standardized data on annual incidence, stroke subtypes, and case fatality rate at 28 days during the years 2005 and 2006. BACKGROUND: An estimated 5.8 million people died from stroke (cerebrovascular disease) in 2005, two thirds of them were from low-/middle-income countries but reliable population-based studies are scarce. METHODS: The manual on WHO STEPwise approach to stroke surveillance (STEPS Stroke; http://www.who.int/chp/steps/Manual pdf) was the operational protocol. We selected a well-defined community (H-district) having verifiable census data and being representative of the population structure of Mumbai (Bombay). Of 337,391 permanent residents, 156,861 persons between the age of 25 and 94+ years who were eligible for survey were screened. The responses to a predefined questionnaire (version 2.0) were entered in coded data sheets for analysis. RESULTS: During the 2-year study period (January 2005 to December 2006), 456 (238 males and 218 females) had FES, indicating an annual incidence in subjects of 25 years and above of 145/100,000 persons (CI 95%: 120-170); for males it is 149/100,000 persons (CI 95%: 120-170) and for females it is 141/100,000 persons (CI 95%: 120-160). The age-standardized rate for study population (both sexes) by the direct method using Segi's 1996 world population is 152/100,000/year (CI 95%: 132-172). Stroke diagnosis was supported by computed tomography in 407 (89.2%) of 456 FES cases: 366 (80.2%) had ischemic stroke, 81 (17.7%) had hemorrhagic stroke and 9 (1.9%) were in the unspecified category. The mean age was 66 +/- (SD) 13.60 years, women were older as compared to men (mean age 68.9 +/- 13.12 years vs. 63.4 +/- 13.53 years). Case fatality: at 28 days, 320 (70%) of 456 FES cases were still alive and 136 (29.8%) had died. Of the 320 surviving patients 38.5% had moderate to severe disability by the modified Rankin scale. CONCLUSIONS: The results of Mumbai stroke study, using uniform definitions and methodologies, show that the annual standardized incidence rates, stroke subtypes and case fatality rate are very similar to those reported from developed nations. To plan effective intervention and prevention strategies, standardized data in representative samples of regional populations are urgently needed.

Exploring of Antimicrobial Activity of Triphala Mashi-an Ayurvedic Formulation.

Triphala Mashi is an ayurvedic formulation that was prepared in our lab. Aqueous and alcoholic extracts of both Triphala and Triphala Mashi were used, to evaluate antimicrobial activity. Comparative phytochemical profile of Triphala and Triphala Mashi was done by preliminary phytochemical screening, total phenolic content and thin layer chromatography (TLC). Antimicrobial activity includes isolation of pathogens from clinical samples, its characterization, testing its multiple drug resistance against standard antibiotics and antimicrobial activity of aqueous and alcoholic extracts of both Triphala and Triphala Mashi against these organisms by using agar gel diffusion method. Triphala Mashi containing phenolic compounds, tannins exhibited comparable antimicrobial activity in relation to Triphala against all the microorganisms tested. It inhibits the dose-dependent growth of Gram-positive and Gram-negative bacteria. In conclusion, it appears that Triphala Mashi has non-specific antimicrobial activity.

Evaluation of anti-diarrhoeal property and acute toxicity of Triphala Mashi, an Ayurvedic formulation.

The anti-diarrhoeal effect of aqueous and alcoholic extracts of Triphala and Triphala Mashi were studied employing castor oil-induced-diarrhoeal model in rats. The gastrointestinal transit rate was expressed as the percentage of the longest distance travelled by the charcoal divided by the total length of the small intestine. All the extracts, at various doses 200, 400 and 800 mg/kg displayed remarkable anti-diarrhoeal activity as evidenced by a significant increase in first defecation time, cumulative fecal weight and intestinal transit time. Aqueous and alcoholic extracts of Triphala and Triphala Mashi were considered safe up to a dose of 1750 mg/kg when evaluated for acute oral toxicity in accordance with the OECD (Organization for Economic Cooperation and Development) guidelines. In conclusion, the remarkable anti-diarrhoeal effect of Triphala and Triphala Mashi extracts against castor oil-induced diarrhoea suggest its potential for application in a wide range of diarrhoeal states.

Role of ultrasonography to detect axillary node involvement in operable breast cancer.

Prompted by the concern about unnecessary axillary dissections, we prospectively studied the accuracy of clinical examination (CE) and conventional ultrasonography (USG, 7.5 MHz), to diagnose pre-operatively metastatic axillary lymph nodes in 200 operable breast cancer patients. USG had higher specificity (90% vs 77%, P = 0.025) and higher positive predictive value (ppv = 90% vs 76%, P = 0.02) than CE. Together, CE + USG had higher sensitivity (82% vs 58%, P = 0.00005) and higher negative predictive value (npv = 76% vs 58%, P = 0.008) than CE alone. In women < 45 years, CE + USG had higher sensitivity (91% vs 76%, P = 0.037) and npv (89% vs 67%, P = 0.018) than in older women. The sensitivity and npv of CE + USG to detect > 1 positive node were 97% (for both) in women < 45 years compared to 81% and 79% in older women. The high sensitivity of CE + USG (82% for the whole group) is probably due to the higher proportion of young women (median age = 45) in our population. It suggests that using CE + USG to avoid axillary dissection in some patients is feasible.

Non-Hodgkin lymphoma of the prostate.

Non-Hodgkin lymphoma of the prostate is rare: only 42 cases have been reported. The authors describe a 27-year-old man with this disease. Lymphoma should be considered as a rare cause of enlargement of the prostate, especially in a young man.

CT staging of pancreatic and periampullary carcinoma.

Sixteen cases of carcinoma head pancreas and seven cases of periampullary carcinoma are staged together on CT scan because of their morphological similarity and similar parameters. Following parameters are considered for CT staging: tumour mass, involvement of splanchnic vessels, locoregional lymph nodes and presence or absence of hepatic metastases. Findings were confirmed on surgical exploration. A contrast enhanced CT scan was 58.3 percent sensitive and 100 percent specific for the involvement of lymph nodes and 100 percent sensitive and 93.4 percent specific for hepatic metastases. The cases diagnosed as non-resectable on CT staging were found inoperable on exploration. Authors believe that for all practical purposes, pancreatic and periampullary malignancies can be grouped together and a contrast enhanced CT scan can provide reliable information for the staging of the tumor.

Degradation of humic acids, extracted from manure and soil by some streptomycetes and fungi.

Humate-decomposing microorganisms were isolated from alluvial soil, farmyard manure and peat by direct plating and enrichment techniques. Peat was found to harbour more of these organisms than soil and manure. Out of 21 actinomycetes and 6 fungi 9 and 3 isolates, respectively, were found to utilize manure-humate as a sole source of carbon and to a less extent as a source of both carbon and nitrogen in liquid medium. These efficient cultures were identified as Streptomyces spec. (A 11 and A 12). Str. candidus, Str. violaceoruber (A 41 and A 42), Str. rubescens, Str. garyphalus, Str. viridochromogenes, Fusarium solani, Aspergillus fumigatus, and Penicillium roseopurpureum. Additional sources of carbon and nitrogen stimulated microbial degradation of humate. Only five isolates formed a zone of decolourization on humate-containing solid medium. Soil humic acid was utilized by Str. candidus as a sole source of carbon and nitrogen, however, F. solani utilized it only in the presence of additional carbon and nitrogen sources. Extent of soil-humate degradation was less than manure-humate.