Multiplexed Imaging Strategy to Distinguish Indeterminant Biliary Strictures: An Ex Vivo Study.

Introduction: Indeterminant biliary strictures can be either malignant or benign. Biliary intraepithelial neoplasia (BilIN) is the precursor lesion to cholangiocarcinoma, a deadly bile duct cancer. Current diagnostic methods are limited by inadequate amounts of cells and tissues collected. Aim: We aim to demonstrate use of fluorescently-labeled peptides specific for EGFR, claudin-1, and ErbB2 to perform multiplexed imaging of biliary neoplasia. Methods: Formalin fixed and paraffin embedded specimens resected from human biliary strictures were sectioned. A gastrointestinal pathologist used standard criteria to score immunohistochemistry from biliary neoplasia and adjacent normal epithelium from the same specimen. Peptides specific for EGFR, claudin-1, and ErbB2 were fluorescently-labeled with FITC, Cy5, and IRDye800, respectively. The fluorophores were chosen to provide spectral separation to distinguish the individual targets. Immuno fluorescence images were collected using confocal microscopy. Results: Target expression was validated using immunohistochemistry. Staining was visualized on the surface of biliary duct epithelial cells and not in the stroma. Greater fluorescence intensity was observed for peptide binding to biliary neoplasia by comparison with normal. The mean ratio for neoplasia-to-normal was 1.4, 1.7, and 1.6, respectively, and the average intensities were significantly greater for neoplasia than normal for each peptide. Peptides and antibody binding co-localized with correlation of ρ=0.64, 0.51 and 0.62, respectively. Conclusions: A panel of fluorescently-labeled peptides can distinguish BilIN and cholangiocarcinoma from normal biliary epithelium, and may be used for multiplexed imaging of indeterminant biliary strictures.

Study on prevalence of ancylostomosis in dogs at Anand district, Gujarat, India.

AIM: This study was undertaken to derive the prevalence rate of ancylostomosis in dogs by a collection of fecal samples from Anand district. MATERIALS AND METHODS: The fecal samples were collected from the dogs brought to the Hospital of Veterinary College (Teaching Veterinary Clinical Service Complex) and the surrounding areas of Anand district. On the day of collection, fecal samples were collected and brought to the Department of Veterinary Parasitology and processed for standard qualitative examination. The sedimentation technique was used to detect the presence of Ancylostoma spp. eggs in the samples. RESULT: The highest prevalence rate was observed in the month of May (36.66% fecal samples) and the lowest in the month of December (13.79% fecal samples) at Anand district. CONCLUSION: It can be concluded that heavy infection is present in Anand district especially in the season of summer followed by monsoon and the least in winter.

Patho-epidemiological study on Genotype-XIII Newcastle disease virus infection in commercial vaccinated layer farms.

AIM: The present research work was carried out to study the patho-epidemiological aspects of Genotype-XIII Newcastle disease virus (NDV) infection in commercial layer in and around Anand, Gujarat. As the outbreaks have reported in vaccinated flocks, it was felt necessary to study the disease with respect to its changing pathogenicity and relevant aspects. MATERIALS AND METHODS: The study comprised of patho-epidemiology of Newcastle disease (ND) by information collected from different layer farms suffering from the disease in relation to incidence pattern and mortality, duration of mortality, susceptible age, and loss due to production performance. Clinical signs were recorded based on observations. During post-mortem, gross lesions were also recorded. For histopathological examination visceral organs according to lesions were collected in 10% formalin and processed slide stained by hematoxylin and eosin for microscopic examination. Cultivation of virus was done in embryonated specific pathogen-free (SPF) eggs of 9-11 days and isolation of virus was done for haemagglutination (HA) and haemagglutination inhibition (HI) test and to identify pathotype of virus by intracerebral pathogenicity index (ICPI) test to determine the virulence of virus. The Genotype-XIII NDV was confirmed by F gene sequence and whole genome sequence. RESULTS: During the study mortality due to ND was recorded in 13 layer flocks in spite of routine vaccination, which usually contain Genotype-II strain of virus. The mortality was observed as high as above 50% with an average of 21.21%. The susceptible age for disease was found to be 6-14 weeks. The duration of mortality observed was 23 days. The disease resulted in a significant reduction in body weight, feed intake and drop in egg production. Majority of the outbreaks appeared during extremely hot months of April to June. Greenish diarrhoea was frequently seen in birds that survived early in infection. Mortality continued for 2-3 weeks and reduced with appearance of torticollis. Gross lesions were characterized by multifocal to diffuse hemorrhages around proventricular glands, necrotic (diphtheritic) haemorrhagic ulcers throughout the intestine, disseminated multiple foci of necrosis and pin-point hemorrhages in the spleen parenchyma. The microscopic lesions include focal to diffuse hemorrhages, diffuse infiltration of mononuclear cells, necrosis, and degeneration in visceral organs. All the 13 farm samples (n=13) resulted in death of all the embryos following incubation up to 72 h post-inoculation. All the 13 allantois fluids from field samples along with F and R2B vaccine sample were found positive for HA activity, which was further confirmed by HI using known NDV serum. The values of ICPI were 2.0 which were indicative of velogenic nature of the field NDV strain. CONCLUSION: The study indicated that presently available live and attenuated vaccines which include Genotype-II NDV have failed in protecting the flocks against Genotype-XIII and resulted in outbreaks with mortality above 50%. ICPI score of 2.0 confirmed that the present outbreaks were due to Genotype-XIII NDV, which is velogenic in nature.

Microwave-assisted rapid synthesis of methyl 2,4,5-trimethoxyphenylpropionate, a metabolite of Cordia alliodora.

Microwave assisted condensation of asaronaldehyde (2) with malonic acid in piperidine-AcOH provides 2,4,5-trimethoxycinnamic acid (3) in 87% yield within 4 min, which upon further reduction with PdCl2- HCOOH-aq. NaOH gives 3-(2,4,5-trimethoxy)phenyl propionic acid (4) in 88% yield within 3 min. Esterification of 4 with MeOH-H+ gives methyl 2,4,5-trimethoxyphenylpropionate (1), a metabolite of Cordia alliodora, in 94% yield within 3 min (overall 69% yield).

Ultrasound-assisted conversion of toxic beta-asarone into nontoxic bioactive phenylpropanoid: isoacoramone, a metabolite of Piper marginatum and Acorus tararinowii.

Ultrasound-assisted synthesis of bioactive isoacoramone (1), a metabolite of Piper marginatum and Acorus tararinowii, has been achieved by oxidation of toxic beta-asarone (2) with potassium permanganate/copper sulphate/alumina into asaronaldehyde (3) followed by treatment with ethylmagnesium iodide to provide 1-(2,4,5-trimethoxy)phenyl-1-propanol (4) which upon further oxidation with potassium permanganate/copper sulphate afforded 1 in 64% yield (overall 32%). Toxicological evaluation of 1 reveals it to be nontoxic up to 60 mg/kg b.w.

One step conversion of toxic beta-asarone from Acorus calamus into 1-(2,4,5-trimethoxyphenyl)-1,2-dihydroxypropane and asaronaldehyde occurring in Piper clusii.

1-(2,4,5-Trimethoxyphenyl)-1,2-dihydroxypropane (2), a natural phenylpropanoid occurring in Piper clusii, has been synthesized for the first time from toxic beta-asarone (1) of Acorus calamus with osmium tetroxide, while 1 with osmium tetroxide (catalytic amount) in presence of sodium metaperiodate furnished the asaronaldehyde (3) in high yield.