Changes in cecum microbial community in response to total sulfur amino acid (TSAA: DL-methionine) in antibiotic-free and supplemented poultry birds.

The effect of essential total sulfur amino acids (TSAA) like methionine and cysteine on the cecal microbiome of broilers was investigated at 2 different time points (days 21 and 42) of broiler rearing. A total of 360-day-old Cobb male broiler chicks were randomly distributed to 6 dietary treatments in a 2 × 3 factorial arrangement, with 2 levels of antibiotic growth promoters (AGP: 0 and 0.05%) and 3 levels of TSAA (DL-methionine) either for starter (0.7, 0.8, and 0.9%) or finisher chicks (0.52, 0.62, and 0.72%), labeled as diets 1 to 6. Cecal digesta from each replicate (n = 10) were sampled on days 21 and 42. DNA was extracted for the amplification of the V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatic analyses were performed using QIIME, Mothur, and ad hoc tools and functional profiles of the inferred metagenome were analyzed using PICRUST. Statistical difference was determined by 2-way ANOVA and PERMANOVA. Clustering of cecal communities using PCoA showed clear separation of microbial communities based on age (P < 0.05) of birds and between low and medium/ high levels of TSAA (DL-methionine). At day 21, bacterial richness and diversity were higher than at day 42 where Clostridium cluster XI and Lactobacillus were found most abundant. No variability in taxonomic richness at the genus level was observed with AGP and DL-methionine supplementation. Interbird variation for richness was greater at day 42 compared to day 21. The mean fold difference of richness was greater (1.5 mean fold) with diets 1 and 6, suggesting interactive effects of AGP and TSAA (DL-methionine) in the diet. KEGG function profiles calculated by PICRUST suggest that the cecal microbiome increased glycolysis and energy generation correlated with increased dietary TSAA (DL-methionine) supplementation levels during the late broiler growth period (day 42). This study increases our knowledge of microbial dynamics and functions that are relevant to host nutrition and performance that may help us tailoring alternative strategies for raising poultry birds under antibiotic-free conditions.

Secreted frizzled-related protein 2 (SFRP2) expression promotes lesion proliferation via canonical WNT signaling and indicates lesion borders in extraovarian endometriosis.

STUDY QUESTION: What is the role of SFRP2 in endometriosis? SUMMARY ANSWER: SFRP2 acts as a canonical WNT/CTNNB1 signaling agonist in endometriosis, regulating endometriosis lesion growth and indicating endometriosis lesion borders together with CTNNB1 (also known as beta catenin). WHAT IS KNOWN ALREADY: Endometriosis is a common, chronic disease that affects women of reproductive age, causing pain and infertility, and has significant economic impact on national health systems. Despite extensive research, the pathogenesis of endometriosis is poorly understood, and targeted medical treatments are lacking. WNT signaling is dysregulated in various human diseases, but its role in extraovarian endometriosis has not been fully elucidated. STUDY DESIGN, SIZE, DURATION: We evaluated the significance of WNT signaling, and especially secreted frizzled-related protein 2 (SFRP2), in extraovarian endometriosis, including peritoneal and deep lesions. The study design was based on a cohort of clinical samples collected by laparoscopy or curettage and questionnaire data from healthy controls and endometriosis patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: Global gene expression analysis in human endometrium (n = 104) and endometriosis (n = 177) specimens from 47 healthy controls and 103 endometriosis patients was followed by bioinformatics and supportive qPCR analyses. Immunohistochemistry, Western blotting, primary cell culture and siRNA knockdown approaches were used to validate the findings. MAIN RESULTS AND THE ROLE OF CHANCE: Among the 220 WNT signaling and CTNNB1 target genes analysed, 184 genes showed differential expression in extraovarian endometriosis (P < 0.05) compared with endometrium tissue, including SFRP2 and CTNNB1. Menstrual cycle-dependent regulation of WNT genes observed in the endometrium was lost in endometriosis lesions, as shown by hierarchical clustering. Immunohistochemical analysis indicated that SFRP2 and CTNNB1 are novel endometriosis lesion border markers, complementing immunostaining for the known marker CD10 (also known as MME). SFRP2 and CTNNB1 localized similarly in both the epithelium and stroma of extraovarian endometriosis tissue, and interestingly, both also indicated an additional distant lesion border, suggesting that WNT signaling is altered in the endometriosis stroma beyond the primary border indicated by the known marker CD10. SFRP2 expression was positively associated with pain symptoms experienced by patients (P < 0.05), and functional loss of SFRP2 in extraovarian endometriosis primary cell cultures resulted in decreased cell proliferation (P < 0.05) associated with reduced CTNNB1 protein expression (P = 0.05). LIMITATIONS REASONS FOR CAUTION: SFRP2 and CTNNB1 improved extraovarian endometriosis lesion border detection in a relatively small cohort (n = 20), although larger studies with different endometriosis subtypes in variable cycle phases and under hormonal medication are required. WIDER IMPLICATIONS OF THE FINDINGS: The highly expressed SFRP2 and CTNNB1 improve endometriosis lesion border detection, which can have clinical implications for better visualization of endometriosis lesions over CD10. Furthermore, SFRP2 acts as a canonical WNT/CTNNB1 signaling agonist in endometriosis and positively regulates endometriosis lesion growth, suggesting that the WNT pathway may be an important therapeutic target for endometriosis. STUDY FUNDING/COMPETING INTEREST(S): This study was funded by the Academy of Finland and by Tekes: Finnish Funding Agency for Innovation. The authors have no conflict of interest to declare.

Characteristic of low vision patients attending an eye hospital in eastern region of Nepal.

INTRODUCTION: Low vision is an important public health problem. OBJECTIVE: To study the profile of low vision patients in a hospital of Nepal. MATERIALS AND METHODS: Information related to the patients' profile, visual status, ocular disease and, low vision devices prescribed were obtained retrospectively from the records of 1,860 visually- impaired patients, regardless of the cause, presenting to the low vision department of the Biratnagar Eye Hospital, Biratnagar, Nepal, over a period of four years. These patients, after a comprehensive ocular examination, underwent low vision examination by an ophthalmologist and a low vision specialist. RESULTS: Of 1,860 patients, males comprised 1298 (70 %), while 562 (30 %) patients were female. Six hundred and one (32.3%) patients were of less than 20 years while 398(21.4%) were more than 60 years of age. Agriculture (500, 27 %), household work (341, 18 %) and students (308, 17 %) were the common occupations. Retinal diseases were the commonest cause of low vision. They were: macular disorders 408 (22 %), retinitis pigmentosa 226 (12.1 %) and other retinal causes 361 (19.4 %) (diabetic retinopathy, choroidal coloboma, post laser for retinal vasculitis and central retinal/branch retinal vein occlusion, healed macular chorioretinal scar secondary to retinochroiditis and choroiditis). Refractive error 215 (11.5 %), amblyopia 49 (2.6 %), optic atrophy 144 (7.8 %) and microphthalmos 105 (5.6 %) were the other causes. Uncorrected distance visual acuity was between 6/24 and 6/60 in 509 (27.4 %) and between 5/60 and PL in 1,327 (71.3 %) patients. Similarly, near visual acuity with vision better than 2.50 M (N 20) and worse than 2.50 M (N20) was present in 643(34.5%) and 1,217(65.5%) patients. About 67% and 54.5% of the patients had some improvement in their distance and near visual acuity with glasses and low vision aids. Distance spectacles 909 (49 %), near spectacles 106 (5.7 %), hand held magnifiers 78 (4 %) and telescopes 18 (1 %) were the optical devices prescribed. CONCLUSION: The prevalence of low vision is common among the people of the younger and older age groups. Retinal diseases are common among the causes for low vision. Adequate prescription and availability of low vision devices can improve the visual acuity.

Profile of ascites patient admitted in Nepal Medical College Teaching Hospital.

Ascites is one of the frequently encountered problems in internal medicine. Common causes of ascites are portal hypertension including cirrhosis of liver and congestive heart failure, hypoalbuminemia associated with nephrotic syndrome, intra-abdominal malignancy and abdominal tuberculosis. We evaluated 43 patients presented with ascites in Nepal Medical College Teaching Hospital (NMCTH). After history taking, clinical examination, imaging studies and laboratory evaluation alcoholic liver disease and abdominal tuberculosis were diagnosed in 19 and 5 patients restively. Constrictive pericarditis was diagnosed in 2 patients and 2 patients were suffering from HCV related liver disease. Present study revealed alcoholic liver disease as the commonest cause of ascites.

Comparison of fine needle aspiration cytology and histopathology in diagnosing cervical lymphadenopathies.

BACKGROUND: Lymphadenopathies are the most common form of extrapulmonary tuberculosis and tuberculous lymphadenitis is the most common cause of peripheral lymphadenopathy in a developing country like Nepal. The objective of this study is to compare the findings of fine needle aspiration cytology to histopathology in diagnosing cervical lymphadenopathies. METHOD: This is a prospective, longitudinal, comparative study was performed in the Department of ENT and Head and Neck Surgery, TU Teaching Hospital, Kathmandu between August 2007 and January 2009. Patients 13 years of age or older were included. There were 55 patients with cervical lymphadenopathies subjected to both fine needle aspiration cytology (FNAC) and histopathology (HPE). The sensitivity, specificity, false positive, false negative, positive predictive value and negative predictive value were calculated. RESULTS: Of 55 patients, 48 patients had lymphadenopathies of infective origin while 7 patients had metastatic disease. Majority of patients (n=20) were in age group 23-32 years and males outnumbered females. The sensitivity, specificity, false positive, false negative, positive predictive value and negative predictive value of FNAC of lymphadenopathies to diagnose tubercular lymphadenopathies were 80.0%, 100.0%, 0.0%, 20.0%, 100.0% and 82.14% respectively. Similarly, sensitivity, specificity of FNAC of lymphadenopathies to differentiate benign and malignant lesion were 100% each. False positive, false negative, positive predictive value and negative predictive value were 0.0%, 0.0%, 100.0% and 100.0% respectively. Overall correlation of FNAC and HPE was 90.9%. CONCLUSION: FNAC can distinguish tubercular lymphadenitis from reactive and granulomatous lymphadenitis in majority of cases. Similarly, it can distinguish well between benign and malignant lymphadenopathies. It can be used as an initial investigation for routine screening of cervical lymphadenopathies where facilities of histopathology are not available.

Pseudomembranous colitis in a pregnant woman.

Pseudomembranous colitis in association with pregnancy has not been well described in English literature. Recent studies show a drastic increase in the incidence and severity of Pseudomembranous colitis in pregnant women, who were once thought to be at low risk. We report here a case of Pseudomembranous colitis in a young healthy immunocompetent pregnant lady. An early suspicion of this entity with the characteristic appearance of pseudomembranes on colonoscopy and histology confirmed the diagnosis enabling prompt treatment and complete recovery without any serious consequences.

Role of foley's catheter in removing foreign body of oesophagus.

Oesophageal foreign bodies are commonly removed with rigid oesophagoscopy under general anaesthesia, but spherical foreign bodies can also be removed using a foley's catheter under fluoroscopic control without anaesthesia. We present a case of successful removal of a spherical marble from the oesophagus in a four years old girl by a foley's catheter under general anaesthesia.

Enzymatic production of trans-free hard fat stock from fractionated rice bran oil, fully hydrogenated soybean oil, and conjugated linoleic Acid.

Rice bran oil (RBO) was fractionated into 2 phases, solid (S-RBO) and liquid (L-RBO), using acetone at -18 degrees C and the weight yield of each S-RBO and L-RBO was 45.5% and 54.5%, respectively. Then, trans-free hard fat was synthesized from trans-free substrate of S-RBO and fully hydrogenated soybean oil (FHSBO) at different molar ratios (S-RBO : FHSBO; 1 : 1, 1 : 1.5, 1 : 2, and 1 : 3) with Lipozyme TL IM lipase (10% of total substrate). Conjugated linoleic acid (CLA, 20% of total substrate) was used as functional fatty acids for the production of trans-free hard fat. After fatty acid analysis, CLA (12.2% to 14.2%) was found on the triacylglycerol (TAG) backbone of the interesterified products along with stearic (37.6% to 49%), palmitic (15% to 17.9%), and oleic acids (13.3% to 19.2%). The interesterified product contained higher level of saturated fatty acid (62.6% to 70.1%) at sn-2 position. Total tocopherols (alpha-, gamma-, and delta-; 1.4 to 2.6 mg/100 g) and phytosterols (campesterol, stigmasterol, and beta-sitosterol; 220.5 to 362.7 mg/100 g) were found in the interesterified products. From DSC results, solid fat contents of the interesterified products (S-RBO : FHSBO 1 : 1, 1 : 1.5, 1 : 2, and 1 : 3) at 25 degrees C were 23.1%, 27%, 30.1%, and 44.9%. The interesterified products consisted mostly of beta' form crystal with a small portion of beta form. The interesterified product (S-RBO : FHSBO 1 : 1.5) was softer than the physical blend but slightly harder than commercial shortenings as measured by texture analyzer. Thus, trans-free hard fat stock, which may have a potential functionality could be produced with various physical properties.

Oesophageal foreign bodies in children.

Foreign body ingestion is a common occurrence and carries significant morbidity and mortality. Failure to treat foreign bodies immediately can lead to various serious complications. This study was done to identify the types as well as site of foreign body ingested and its complication in children. A retrospective study of 122 cases of suspected foreign body ingestion in patients admitted in ENT and Head and Neck Surgery of TU Teaching Hospital, Kathmandu were done in between April 2004 to July 2008. Ages less than 12 years were included. In all cases x-ray soft tissue neck lateral and chest x-ray posterio-anterior views were done along with other preoperative investigations. Rigid oesophagoscopy or hypopharyngoscopy were done under general anesthesia to remove foreign bodies. There were 64.7% male and 35.3% female children. Foreign bodies were common in 0-4 year age group. Most common foreign body were coin (64.0%) followed by meat bone (14.0%). No foreign bodies were found in 2.4% patients as they were passed in stomach. No complications were noted during the entire period of this study. Most common foreign bodies in children are coin. Though complications with these foreign bodies are rare, these do occur due to delay in presentation and removal. No complications were noted in our series. Eventhough children who swallow foreign bodies are asymptomatic; we must maintain a high index of suspicion and undergo diagnostic procedure, if there is a positive history.

Phytoextraction of zinc, copper, nickel and lead from a contaminated soil by different species of Brassica.

In a pot culture experiment, five different species of Brassica (Brassica juncea, Brassica campestris, Brassica carinata, Brassica napus, and Brassica nigra) were grown for screening possible accumulators of heavy metals, viz. Zn, Cu, Ni, and Pb. The plants were grown to maturity in a soil irrigated with sewage effluents for more than two decades in West Delhi, India. The soil analysis showed enhanced accumulation of Zn, Cu, Ni, and Pb in this sewage-irrigated soil. Among all species, B. carinata showed the highest concentration (mg kg(-1)) as well as uptake (microg pot(-1)) of Ni and Pb at maturity. Although B. campestris showed a higher concentration of Zn in its shoots (stem plus leaf), B. carinata extracted the largest amount of this metal due to greater biomass production. However, B. juncea phytoextracted the largest amount of Cu from the soil. In general, the highest concentration and uptake of metal was observed in shoots compared to roots or seeds of the different species. Among the Brassica spp., B. carinata cv. DLSC1 emerged as the most promising, showing greater uptake of Zn, Ni, and Pb, while B. juncea cv. Pusa Bold showed the highest uptake of Cu. The B. napus also showed promise, as it ranked second with respect to total uptake of Pb, Zn, and Ni, and third for Cu. Total uptake of metals by Brassica spp. correlated negatively with available as well as the total soil metal concentrations. Among the root parameters, root length emerged as the powerful parameter to dictate the uptake of metals by Brassica spp. Probably for the first time, B. carinata was reported as a promising phytoextractor for Zn, Ni, and Pb, which performed better than B. juncea.

Sphenoid sinus mucocele with blindness: a rare presentation.

Isolated sphenoid sinus mucocele is a rare entity. Here we report a case of isolated sphenoid sinus mucocele in a 9 years old girl presenting with unilateral blindness and without any sinonasal complaints. C.T. scan revealed an isolated sphenoid sinus mucocele which was managed endoscopically. A brief review of literature is also included.

Myiasis infestation in postoperative mastoid cavity.

This is a case report of myiasis in modified radical mastoidectomy cavity in a 48 years old male presented in the Department of ENT and Head and Neck Surgery, T.U. Teaching Hospital, Katmandu, Nepal. Around 50 maggots were removed from his mastoid cavity. Patient was managed with turpentine oil and antibiotics which was given to prevent secondary infection. Literature search revealed only one case report of Myiasis infestation in a postoperative mastoid cavity.

Fine Needle Aspiration cytology versus Fine Needle Sampling without aspiration. A prospective study of 200 cases.

In the present prospective study of 200 patients with superficial swelling of various sites, FNS followed by FNA were performed. Fifteen cases were excluded due to inadequate material. In the 80 lymph node cases, FNS was diagnostic in 85% cases as compared to FNA in 87.5%. The diagnostic accuracy was higher for FNA (87.5%) than for FNS (81.25%). However FNS smears were much superior qualitatively than FNA smears and for the diagnosis of malignant conditions FNS was found to be much better. Regarding the 68 thyroid swellings, FNS obtained a diagnosis in 82.4% cases while FNA got it in 77.9%. The diagnostic accuracy with FNS (89%) was much better compared to FNA (75%). Of the 27 breast cases FNS yielded diagnostic material in 70.38% cases while FNA was diagnostic in 85.19%, thus establishing definite superiority. However FNS seemed to be better for diagnosing malignant lesions while FNA appeared better for diagnosing benign ones. In the 5 salivary gland lesions and 5 miscellaneous lesions both FNS and FNA fared equally well. On the whole FNS technique was much more patient friendly and gave high class "text book" quality smears while FNA smears gave quantitatively more adequate material. Both the techniques therefore would be complementary to each other.

Mutational analysis at Asn-41 in peanut agglutinin. A residue critical for the binding of the tumor-associated Thomsen-Friedenreich antigen.

Peanut agglutinin is a clinically important lectin due to its application in the screening of mature and immature thymocytes as well as in the detection of cancerous malignancies. The basis for these applications is the remarkably strong affinity of the lectin for the tumor-associated Thomsen-Friedenreich antigen (T-antigen) and more so due to its ability to distinguish T-antigen from its cryptic forms. The crystal structure of the complex of peanut agglutinin with T-antigen reveals the basis of this specificity. Among the contacts involved in providing this specificity toward T-antigen is the water-mediated interaction between the side chain of Asn-41 and the carbonyl oxygen of the acetamido group of the second hexopyranose ring of the sugar molecule. Site-directed mutational changes were introduced at this residue with the objective of probing the role of this residue in T-antigen binding and possibly engineering an altered species with increased specificity for T-antigen. Of the three mutants tested, i.e. N41A, N41D, and N41Q, the last one shows improved potency for recognition of T-antigen. The affinities of the mutants can be readily explained on the basis of the crystal structure of the complex and simple modeling. In particular, the change of asparagine to glutamine could lead to a direct interaction of the side chain with the sugar while at the same time retaining the water bridge. This study strengthens the theory that in lectins the nonprimary contacts generally made through water bridges are involved in imparting exquisite specificity.

Upstream flanking sequences and transcription of SINEs.

SINEs, short interspersed repeated DNA elements, undergo amplification through retroposition and subsequent integration into a new location in the genome. Each new SINE insertion will be located in a new chromosomal environment, with different flanking sequences. Modulation of transcription by different flanking sequences may play an important role in determining which SINE elements are preferentially active in a genome. We evaluated the ability of upstream flanking sequences to regulate the transcription of three different SINEs (Alu, B2 and ID) by constructing chimeric constructs with known 5' flanking sequences of RNA polymerase III-transcribed genes. Upstream sequences from the 7SL RNA gene, U6 RNA gene, vault RNA gene, and BC1 gene increase transcription of Alu, B2 and BC1 in transient transfections of NIH3T3, HeLa, Neuro2a and C6 glioma cell lines. The 7SL sequence proved most efficient in increasing SINE transcription. The 7SL upstream fused to the BC1 RNA gene (an ID element) was used to create a transgenic mouse line. In contrast to the tissue-specific endogenous BC1 transcription, BC1 transgene transcripts were detected in all tissues tested. However, expression was much higher in those tissues that express the endogenous gene, demonstrating both transcriptional and post-transcriptional regulation. The BC1 RNA was detected in a similar ribonucleoprotein complex in the different tissues.

Fine needle aspiration cytology in systemic lupus erythematosus lymphadenopathy. A case report.

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown etiology characterized by inflammation in various organ systems, including lymph nodes, due to the production of antinuclear antibodies. The onset of disease is between ages 13 and 40 years, with a female preponderance. CASE: A 30-year-old female presented with right cervical lymphadenopathy and gave a history of intermittent fever and swollen joints of 2.5 years' duration. The patient was on intermittent corticosteroids. With a suggestion of tuberculous lymphadenitis, the patient underwent fine needle aspiration (FNA). The diagnosis of lupus adenopathy was established by FNA of enlarged right cervical lymph nodes. Smears showed predominantly typical and atypical immunoblasts, plasma cells, occasional Reed-Sternberg-like cells and dispersed hematoxylin bodies. Smears were negative for acid-fast bacilli. CONCLUSION: When SLE patients develop lymphadenopathy, FNA cytology helps differentiate lupus adenopathy from infectious conditions, such as tuberculous adenitis, and from Kikuchi's lymphadenitis.

FNAC diagnosis of tuberculosis--an eight years study at Mangalore.

1. FNAC is a very useful adjunct in the diagnosis of tuberculosis: 2. Diagnosis of tuberculosis can be made by the demonstration of epithelioid granulomas with or without caseation even in the absence of AFB. 3. Necrotic features whether acellular or accompanied by neutrophilic infiltrate are usually misdiagnosed as suppurative abscesses. Such smears however show high AFB positivity and thus the diagnosis of tuberculosis is still possible even in the absence of epithelioid granulomas. 4. Therefore all smears obtained by FNAC of suspected tuberculous lesions should be subjected to ZN staining for AFB.