Molecular Characterization of a Copper Metallothionein Gene From a Ciliate Tetrahymena farahensis.

A new copper metallothionein (TfCuMT) gene has been identified from a locally isolated ciliate Tetrahymena farahensis. It contains 327 nucleotides encoding a peptide chain of 108 amino acids and belongs to class MTT2 and subfamily 7b. Amplification from both gDNA and mRNA confirmed the intronless nature of this gene. Like most of the metallohtioneins, cysteine residues contribute nearly 30% content with the specific CKC motifs. Structural repeats present in peptide sequence of TfCuMT indicate internal duplication of gene at some stage of gene evolution. The predicted model of copper metallothionein protein showed that copper ions are mainly chelated by thiol sulfur of cysteine residues and are embedded in the folds of polypeptide chain. For in vivo expression of TfCuMT in Escherichia coli host cells the classical stop codons, which coded for glutamine in the ciliate were mutated to CAA and CAG through site directed mutagenesis. The mutated gene showed higher expression in pET28a expression vector compared with pET21a. Optimum expression was obtained after 6-8 h of 0.1 mM IPTG induction. Stability of His tagged TfCuMT in 5% SDS was low, with half-life of about 104 min. Presence of 1.0 µM copper increased the expression level by 1.65-fold. Presence of 100 µM Cysteine in culture medium caused 2.4-fold increase in expression level. His tagged TfCuMT was purified through affinity chromatography using NTN-His binding resin in the presence of 0.1 M imidazole and NaCl. The modeled structure of the TfCuMT showed a cleft for Cu binding with correct orientation of Cys residues in the motif CKC. J. Cell. Biochem. 117: 1843-1854, 2016. © 2016 Wiley Periodicals, Inc.

Mapping QTLs for swimming ability related traits in Cyprinus carpio L.

Body height (BH), head length (HL), snout length (SL), and tail length (TL) are important traits related with swimming ability of fish. Therefore, improving these traits will increase the production which is the basic goal of aquaculture breeding. To understand the genetic basis of swimming ability related traits in Cyprinus carpio L., a high-density linkage map spanning 3,301 cM in 50 linkage groups was utilized for quantitative trait locus (QTL) mapping. Mapping family comprised 190 offspring and 627 molecular markers were genotyped with average distance of 5.6 cM. A total of 15 QTLs including four (qBH13, qBH30, qBH33, qBH48) for BH, four (qHL10, qHL18, qHL29, qHL48) for HL, three (qSL24, qSL27, qSL45) for SL, and four (qTL15, qTL17, qTL18, qTL44) for TL were detected on 13 linkage groups LG10, LG13, LG15, LG17, LG18, LG24, LG27, LG29, LG30, LG33, LG44, LG45, and LG48. Each LG consisted on single QTL except LG18 and LG48. LG18 was found with two QTLs associated with HL and TL. While LG48 was comprised, the QTLs related with BH and HL. The phenotype variance was recorded from 12.6 to 40.6 %. Five QTLs, qHL48, qSL45, qTL15, qTL18, and qTL44, explained phenotype variance of >20 % with a significant levels of 0.047, 0.049, 0.037, 0.025, and 0.023, respectively. The neighbored loci of these QTLs were considered as main region of chromosomes controlling the traits. These identified genetic regions will be the main source of discovering gene(s) associated with swimming ability related traits in C. carpio L.

Do patients at risk of sleep apnea have an increased risk of cardio-respiratory complications during endoscopy procedures?

BACKGROUND: Patients with sleep apnea (OSA) have an increased risk of perioperative complications. AIM: The purpose of this study is to assess whether OSA increases the risk of cardio-respiratory complications in patients undergoing endoscopic procedures with conscious sedation. METHODS: A prospective study over a 7-month period was performed. All patients undergoing upper, lower, or combined endoscopy were asked to fill in the Berlin questionnaire. The questionnaire was scored, and patients were classified as high or low risk for sleep apnea based on the suggested scoring criteria. Patients who had previously undergone a sleep study were excluded. Demographics and co-morbidities were identified from the electronic medical record. Procedure type, amount of sedation, and minor and major complications were identified from the endoscopy flow sheet. The minor complications were defined as hypertension, hypotension, bradycardia, tachycardia, hypoxemia, and bradypnea (respiratory rate <8 breaths/min). Major complications included chest pain, arrhythmia, altered mental status, respiratory distress, and a minor complication that required a significant intervention, such as use of a reversal agent, atropine, up-titration of oxygen for hypoxemia, or prolonged observation. RESULTS: Procedures were performed in 904 patients: colonoscopies, 68.0%; upper endoscopies, 22.8%; and combined procedures, 9.2%. Five hundred fifty-three patients were identified as low risk (61.2%), and 351 were identified as high risk (38.8%). The mean age was 59.5 ± 10.5 years, mean body mass index was 28.9 ± 6.6, mean neck circumference was 16.2 ± 6.3 in., and 91.4% were males. The median Charlson co-morbidity index was 1 (25-75% percentage range 0-2). All patients received midazolam and fentanyl during endoscopy. The median and 25-75% range for midazolam and fentanyl dosages were 5 mg, 4-6 mg and 100 µg, 75-125 µg, respectively. Minor complications were observed in 10.56% of low-risk patients and 10.63% of high-risk patients (p = not significant (NS); odds ratio, 1.01; 95% confidence interval 0.65-1.56). Major complications were observed in 3.25% of low-risk patients and 1.9% of high-risk patients (p = ns; odds ratio, 0.6; 95% confidence interval 0.26-1.46). CONCLUSION: For patients undergoing endoscopy procedures under conscious sedation, the presence of OSA does not clearly increase the risk of cardiopulmonary complications.

A novel approach to paraneoplastic intestinal pseudo-obstruction.

Paraneoplastic neurologic syndromes (PNS) are uncommon, affecting fewer than 1 in 10,000 patients with cancer. PNS, while rare, can cause significant morbidity and impose enormous socio-economic costs, besides severely affecting quality of life. PNS can involve any part of the nervous system and can present as limbic encephalitis, subacute cerebellar ataxias, opsoclonus-myoclonus, retinopathies, chronic intestinal pseudo-obstruction (CIPO), sensory neuronopathy, Lambert-Eaton myasthenic syndrome, stiff-person syndrome, and encephalomyelitis. The standard of care for CIPO includes the use of promotility and anti-secretory agents and the resection of the non-functioning gut segment; all of which can cause significant compromise in the quality of life. There is significant evidence that paraneoplastic neurologic syndromes are associated with antibodies directed against certain nerve antigens. We successfully treated a patient with CIPO in the setting of small cell lung cancer with a combination of rituximab and cyclophosphamide. The patient, who had failed to respond to prokinetic agents, anti-secretory therapy, and multiple resections, responded to the immunomodulatory therapy, with minimal residuals with PEG tube feeding and sustained ostomy output. The use of rituximab and cyclophosphamide should therefore be considered in patients with CIPO, especially if it can avoid complicated surgical procedures.

Does sleep apnea increase the risk of cardiorespiratory complications during endoscopy procedures?

BACKGROUND: Patients with obstructive sleep apnea (OSA) have an increased risk of perioperative complications. AIM: The purpose of this study is to assess whether OSA increases the risk of cardiorespiratory complications in patients undergoing endoscopic procedures. METHODS: A retrospective study was performed. We identified all patients who had undergone both an endoscopic procedure under conscious sedation and a sleep study from January 2001 to May 2008. Patients were divided into four groups: OSA negative (apnea-hypopnea index (AHI) < 5/h), OSA positive; mild: AHI 5-15/h, moderate: AHI 15.1-30/h, and severe: AHI > 30/h. Minor and major complications were identified. The minor ones were hypertension, hypotension, bradycardia, tachycardia, oxygen desaturation (<90%), and bradypnea. Major complications included chest pain, respiratory distress, cardiorespiratory arrest, or any minor complication that required intervention. RESULTS: Procedures were performed in 639 patients: colonoscopies 68.5%, upper endoscopies 20.2%, and combined procedures 11.3%. The mean age was 60.5 years, mean body mass index 33.7, and 93% were males. Sleep study results: 130 negative, 509 positive; 135 mild, 125 moderate, and 249 severe. Of the patients, 19% had minor complications, while 7% had major complications. There was no significant difference between the patients with and without OSA in the rate of minor complications (odds ratio 1.17, 95% confidence interval 0.70-1.92) or major complications (odds ratio 1.19, 95% confidence interval 0.54-2.63). The odds ratio was also not significantly increased when a cutoff value of 10 or 15/h was used to delineate a positive sleep study. CONCLUSION: For patients undergoing endoscopy procedures under conscious sedation, the presence of OSA does not clearly increase the risk of cardiorespiratory complications.

Quality index of oils extracted from gamma-irradiated peanuts (Arachis hypogaea L.) of the golden and bari varieties.

Two varieties of peanuts were irradiated to 4, 6 and 8 kGy with Co(60). Their proximate compositions remained unaffected, but microbes were eliminated completely after irradiation to 8 kGy. HPLC was used to study tocopherols of irradiated and unirradiated oil samples. There were dose-dependent differences in physico-chemical values between the control and irradiated samples. Significant changes in tocopherol concentrations and peroxide values in the oils were observed after irradiation to 8 kGy. Fatty acid compositions did not change significantly. The study has shown that irradiation is an effective tool in preservation of peanut oil.

Mi-DISCOVERER: A bioinformatics tool for the detection of mi-RNA in human genome.

MicroRNAs (miRNAs) are 22 nucleotides non-coding RNAs that play pivotal regulatory roles in diverse organisms including the humans and are difficult to be identified due to lack of either sequence features or robust algorithms to efficiently identify. Therefore, we made a tool that is Mi-Discoverer for the detection of miRNAs in human genome. The tools used for the development of software are Microsoft Office Access 2003, the JDK version 1.6.0, BioJava version 1.0, and the NetBeans IDE version 6.0. All already made miRNAs softwares were web based; so the advantage of our project was to make a desktop facility to the user for sequence alignment search with already identified miRNAs of human genome present in the database. The user can also insert and update the newly discovered human miRNA in the database. Mi-Discoverer, a bioinformatics tool successfully identifies human miRNAs based on multiple sequence alignment searches. It's a non redundant database containing a large collection of publicly available human miRNAs.

Extended octreotide suppression test to determine hormone responsiveness of multiple type I gastric carcinoid tumors.

A 70-year-old man was found to have at least 12 type I gastric carcinoids and microcarcinoidosis. We performed an extended octreotide suppression test to determine if the tumors were gastrin-dependent and would likely regress after antrectomy. He was given an octreotide infusion at 12.5-25 mcg/h for 86 hr followed by depot octreotide 20 mg intramuscularly every four weeks for eight months. Fasting serum gastrin and chromogranin A levels were measured, and endoscopy with biopsies was performed before and after the infusion and at five months and eight months. Total RNA was extracted for quantitation of histidine decarboxylase mRNA using real-time PCR. Fasting serum gastrin decreased from 306 pg/ml pretreatment to 31 pg/ml at the end of infusion and 115 pg/ml at eight months. Chromogranin A decreased from four to six times the upper limit of normal to normal. Tissue histidine decarboxylase mRNA decreased 50-fold. At eight months, only a few diminutive nodules were present on endoscopy. These results demonstrated that the carcinoid tumors in this patient were under neuroendocrine control and were expected to respond to antrectomy.

Is blinding the endoscopists to bowel preparations in randomized-controlled trials a reality?

BACKGROUND: A number of studies compared the quality, efficacy and tolerability of oral sodium phosphate (NaP) and polyethylene glycol (PEG)-based solutions in preparation for colonoscopy. The primary aim of this study was to explore whether endoscopists can be effectively blinded to the type of bowel preparation. METHODS: We recruited 3 experienced endoscopists and 57 outpatients (18-65 years old) undergoing colonoscopy. We randomized eligible patients to receive one of the two bowel preparations. Endoscopists who performed the tests were blinded to the type of preparation, and made their best judgment on the type and quality of the bowel preparation. RESULTS: Forty-five patients completed the study. The overall correct estimation of the type of bowel preparation was 60.0% (95% CI; 45.5%, 73.0%). The cleansing quality did not differ between the two preparations. Patients found oral NaP solution much easier to take (81.8% versus 36.4%; P = 0.005) and the PEG-based group tended to have more nausea or vomiting. 47.6% of patients in the PEG group indicated they would prefer to try another bowel preparation in the future compared to 4.5% in the oral NaP group (P = 0.002). We stopped the study after an interim analysis indicating that more than 600 patients would be required to detect statistically significant differences in the primary aim. CONCLUSION: Our findings suggest that blinding of endoscopists in clinical trials comparing oral NaP to PEG had a relatively low likelihood of bias. The study also suggests that oral NaP is easier to take and more tolerable than PEG without impairing cleansing quality.

Systemic sarcoidosis presenting as a granulomatous tattoo reaction secondary to interferon-alpha treatment for chronic hepatitis C and review of the literature.

Sarcoidosis is a multisystem granulomatous disease of unknown etiology. Immune alterations involving heightened T-helper-1 responses have been proposed to play a major role in the pathogenesis of sarcoidosis. Interferon-alpha therapy and hepatitis C infection have been implicated in the development of a variety of autoimmune diseases. However, despite the wide use of IFN-alpha therapy for hepatitis C, only a few cases of sarcoidosis have been reported in this context. We report the case of a 42-year-old white female with hepatitis C, who developed systemic sarcoidosis shortly after therapy with IFN-alpha2b. The disease was heralded by the appearance of a cutaneous sarcoid/ foreign body granulomatous reaction at the site of an old tattoo. The sarcoidosis responded to a short course of oral prednisone therapy. We also reviewed the other reported cases and discussed the possible immunological mechanisms involved.