Cryopreservation for delayed circulating tumor cell isolation is a valid strategy for prognostic association of circulating tumor cells in gastroesophageal cancer

AIM To demonstrate the feasibility of cryopreservation of peripheral blood mononuclear cells(PBMCs) for prognostic circulating tumor cell(CTC) detection in gastroesophageal cancer.METHODS Using 7.5 m L blood samples collected in EDTA tubes from patients with gastroesopheagal adenocarcinoma, CTCs were isolated by epithelial cell adhesion molecule based immunomagnetic capture using the Iso Flux platform. Paired specimens taken during the same blood draw(n = 15) were used to compare number of CTCs isolated from fresh and cryopreserved PBMCs. Blood samples were processed within 24 h to recover the PBMC fraction, with PBMCs used for fresh analysis immediately processed for CTC isolation. Cryopreservation of PBMCs lasted from 2 wk to 25.2 mo(median 14.6 mo). CTCs isolated from pre-treatment cryopreserved PBMCs(n = 43) were examined for associations with clinicopathological variables and survival outcomes.RESULTS While there was a significant trend to a decrease in CTC numbers associated with cryopreserved specimens(mean number of CTCs 34.4 vs 51.5, P = 0.04), this was predominately in samples with a total CTC count of 50, with low CTC count samples less affected(P = 0.06). There was no significant association between the duration of cryopreservation and number of CTCs. In cryopreserved PBMCs from patient samples prior to treatment, a high CTC count( 17) was associated with poorer overall survival(OS)(n = 43, HR = 4.4, 95%CI: 1.7-11.7, P = 0.0013). In multivariate analysis, after controlling for sex, age, stage, ECOG performance status, and primary tumor location, a high CTC count remained significantly associated with a poorer OS(HR = 3.7, 95%CI: 1.2-12.4, P = 0.03). CONCLUSION PBMC cryopreservation for delayed CTC isolation is a valid strategy to assist with sample collection, transporting and processing.     

Calcitonina como agente analgésico: revisão dos mecanismos de ação e das aplicações clínicas

Background and objectivesCalcitonin is a polypeptide hormone regulating the metabolism calcium in the body. For many years calcitonin has been used to maintain and improve bone mineral density and to reduce the fracture rate. Many studies showed that calcitonin had analgesic role in several painful circumstances. This pain‐ameliorating effect is irrelevant to its osteoclastic inhibitory effect and mechanisms like altering Na+ channel and serotonin receptor expression or hypothesis including the endorphin‐mediated mechanism were used to explain this effect. In this study we performed a thorough review on the role of calcitonin as an analgesic agent in different scenarios and investigated the fact that calcitonin can be a feasible medication to relieve pain.MethodMany studies focused on the analgesic effect of calcitonin in several painful circumstances, including acute pains related to vertebral fractures, metastasis, migraine and reflex sympathetic dystrophy as well as neuropathic pains related to spinal injuries or diabetes, and phantom pain. Also, calcitonin was showed to be a useful additive to local anesthesia in the case of controlling postoperative pain or trigeminal neuralgia more effectively. However we faced some contradictory data for conditions like lumbar canal stenosis, complex regional pain syndrome, phantom pain and malignancies.ConclusionThis study showed that calcitonin could be helpful analgesic agent in different painful situations. Calcitonin can be considered an eligible treatment for acute pains related to vertebral fractures and a feasible alternative for the treatment of the acute and chronic neuropathic pains where other medications might fail.     

Mutation screening of TGFBI in two Iranian Avellino corneal dystrophy pedigrees

Purpose: Genetic analysis and phenotypic features of Avellino corneal dystrophy patients from Japan and some European countries have been published. We report for the first time the genetic analysis and phenotypic features of two Avellino corneal dystrophy pedigrees from the Middle East. Methods: Slit‐lamp biomicroscope photographs of cornea were obtained, and corneal tissue sections were stained with masson‐trichrome and Congo red. DNA was isolated from peripheral blood leucocytes and exons 4 and 12 of TGFBI were screened for mutations by direct sequencing. Results: The probands of the pedigrees had phenotypic features consistent with diagnosis of Avellino corneal dystrophy. They were homozygous for the same R124H mutation in TGFBI as previously reported in Avellino patients from Japan and European countries. Heterozygous carriers of the mutation were identified in the pedigree and shown to have symptoms of disease milder than those of the probands. Conclusion: The finding of R124H in the Middle Eastern (Iranian) population supports the proposal that perhaps only substitution of histidine for arginine at position 124 of tumour growth factor beta induced protein results in the Avellino corneal dystrophy phenotype. As both probands were originally diagnosed with granular corneal dystrophy, and as heterozygous carriers of R124H were unaware of their disease status prior to genetic analysis, the importance of genetic analysis is emphasized.     

Genetic analysis and SOD1 mutation screening in Iranian amyotrophic lateral sclerosis patients

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease, and the most common in European populations. Results of genetic analysis and mutation screening of SOD1 in a cohort of 60 Iranian ALS patients are here reported. Initially, linkage analysis in 4 families identified a disease-linked locus that included the known ALS gene, SOD1. Screening of SOD1 identified homozygous p.Asp90Ala causing mutations in all the linked families. Haplotype analysis suggests that the p.Asp90Ala alleles in the Iranian patients might share a common founder with the renowned Scandinavian recessive p.Asp90Ala allele. Subsequent screening in all the patients resulted in identification of 3 other mutations in SOD1, including p.Leu84Phe in the homozygous state. Phenotypic features of the mutation-bearing patients are presented. SOD1 mutations were found in 11.7% of the cohort, 38.5% of the familial ALS probands, and 4.25% of the sporadic ALS cases. SOD1 mutations contribute significantly to ALS among Iranians.     

Isolation of Small Number of Cryptosporidium parvum Oocyst Using Immunochromatography

Background

Cryptosporidium parvum causes severe gastroenteritis in immunocompromised human and new borne animals. The organism can be transmitted through water. Since small number of C. parvum is infectious, the aim of the present study was to develop a chromatography method for the isolation of C. parvum oocyst in samples with limited number of oocysts.

Methods

Antibody was prepared against whole antigen from C. parvum oocysts, the achieved Ab bound to the sepharose 4B and used for the isolation of oocysts. Antibody against P23 bound to the sepharose 4B, used also for the isolation of C. parvum oocyst. In comparison to these both methods, 2 traditional methods (Salt floatation and 55% sucrose floatation) were also performed.

Results

Both chromatography methods could bind oocysts with capacity depends on the column size. The isolated oocysts were free of bacteria. Our results showed that the traditional methods are useful for the isolation of oocysts from feces, in its smear stained with ziehl-nelsen, at least 3 oocyts are detectable in each microscopic field under 1000 X magnification. In contrast to the chromatography methods, the bacterial contamination was always observed in oocysts isolated with traditional methods.

Conclusion

Immunochromatography could be used for the successful isolation of C. parvum oocysts from the samples containing limited number of oocysts.     

Cytokine Responses of Human Blood Monocytes Stimulated with Igs

Using an in vitro system for stimulating human peripheral blood mononuclear cells (PBMC) with immobilized Ig, patterns of cytokine production as a function of different Ig classes and subclasses were elucidated. Wells were coated with IgA, IgG1, IgG2, IgG3 or IgG4. Equivalent protein content on surfaces of wells was demonstrated by a human kappa chain ELISA. Isolated human PBMC were added to Ig-coated wells and incubated for 24 hrs before supernatants were assayed for cytokines. The IgG subclasses showed differences in cytokine production stimulated from PBMC, with the relative stimulation for TNF being IgG2 IgG3 IgG1 > IgG4 and for IL-6 production, IgG2 IgG3 > IgG1 = IgG4. In contrast, the relative stimulation for IL-8 was IgG1 = IgG2 = IgG3 = IgG4. IgA caused less production of TNF when compared to IgG2, but similar levels of IL-8. Such differences may have important implications in the pathogenesis of immune complex mediated diseases.     

Hesperidin improves hepatic steatosis,hepatic enzymes,and metabolic and inflammatory parameters in patients with nonalcoholic fatty liver disease: A randomized,placebo‐controlled,double‐blind clinical trial

This study aimed to evaluate the effects of hesperidin on nonalcoholic fatty liver disease (NAFLD) characteristics. In this randomized, double‐blind, controlled clinical trial, 50 NAFLD patients were supplemented with either 1‐g hesperidin capsule or identical placebo capsule for 12 weeks. During the intervention, both groups were advised to follow healthy lifestyle habits including dietary and physical activity recommendations. At the end of the study, hesperidin supplementation, compared with placebo, was associated with a significant reduction in alanine aminotransferase (p = .005), γ‐glutamyltransferase (p = .004), total cholesterol (p = .016), triglyceride (p = .049), hepatic steatosis (p = .041), high‐sensitivity C‐reactive protein (p = .029), tumor necrosis factor‐α, and nuclear factor‐κB (NF‐κB). In conclusion, our results indicate that hesperidin supplementation accompanied with lifestyle modification is superior to lifestyle modification alone in management of NAFLD at least partially through inhibiting NF‐κB activation and improving lipid profile. Further studies with higher dose of hesperidin are required to find the optimal dose.     

Demographic background and dangerous behaviors of heroin-addicted patients under maintenance treatment in the North East of Iran (North Khorasan province)

Background: In recent years, Iran has experienced an increase in heroin consumption, which endangers the health of individuals as well as the whole society. The present research investigated the demographic indices and dangerous behavior of heroin-addicted patients under maintenance treatment. Methods: This study was conducted as a cross-sectional study in 2015. The sample of this research includes the heroin-addicted patients under treatment in 79 maintenance treatment centers in North Khorasan. The information is gathered by a checklist of dangerous behaviors and demographic indices of the patients. Results: In this study, 842 heroin consumers under maintenance treatment were investigated, 82.4% of whom were male and 17.6% were female. Heroin-addicted patients showed a significant difference in terms of marital status, occupation, addiction initiation, and dangerous behaviors (such as unsafe sexual intercourse, the history of concurrency injection and unsafe sexual intercourse) in two gender groups. Conclusion: It seems that there is a significant difference between male and female heroin-addicts in dangerous behaviors and demographic indices. As such, there is a significant difference between demographic pattern and dangerous behaviors of heroin-addicted patients in Iran and patients from other countries.