Association of vitamin D genetic pathway with asthma susceptibility in the Kurdish population.

BACKGROUND: Vitamin D (Vit D) function in asthma progression has been studied well. The effects of genetic variations in Vit D pathway molecules have been also studied, although the results are contradicted. In the present study, for the first time we examined the Vit D pathway molecules included serum Vit D and vitamin D-binding protein (VDBP) and also genetic variations in the vitamin D receptor (VDR) and VDBP in a Kurdish population with asthma. METHODS: An enzyme-linked immunosorbent assay (ELISA) method was used to measure the serum Vit D and VDBP. VDR rs1544410 and rs2228570 and VDBP rs7041 were assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The serum level of Vit D significantly decreased in asthmatic patients versus controls (16.26 ± 6.76 vs 23.05 ± 10.57 ng/mL, P value = .001). We observed an indirect correlation between Vit D and clinical findings. We also found an increased level of serum VDBP in patients as compared to the controls (1044.6 ± 310.82 vs 545.95 ± 121.73 µg/mL, P value < .0001). Besides, the risk of asthma progression was increased in patients with the VDR rs2228570 CC and VDBP rs7041 GG genotypes (OR = 3.56, P = .0382 and OR = 2.58, P = .01, respectively). CONCLUSION: In summary, our results explain the influence of the genetic variations in VDR and VDBP in addition to Vit D and VDBP serum concentrations on asthma susceptibility in the Kurdish population.

A seroprevalence and relationship survey of brucellosis between pregnant women and women with spontaneous abortion in Iran.

Background: Brucellosis is one of the most prevalent diseases common between humans and animals. It is also called Malta fever, Undulant fever and Mediterranean fever. This disease is spread by consuming milk and its unpasteurized derivatives. Clinical symptoms of brucellosis in humans are fever, chills, headache, muscular pain, tiredness, loss of appetite, joint pain, weight loss, constipation, sore throat, and dry cough. The present study aimed at surveying the seroprevalence of brucellosis in pregnant women and those women who suffered from spontaneous abortion. Methods: This case- control study was conducted in Sanandaj (Iran) in 2016 and included 2 groups of pregnant women: one group included 160 pregnant women and the other included 160 women who suffered from spontaneous abortion. Then, the participants were asked to fill out the questionnaire. After receiving permission from an obstetrician, a 10-cc blood sample was taken from each person to be used in the Rose Bengal, Wright, 2ME, and Coombs tests. Independent samples t test and Chi-square test were used to analyze the data and compare the groups. Results: Mean±SD age of women in the case group was 30.9±7.3 years, while it was 27.74±5.41 years in control women. The Rose Bengal, Wright, and 2ME prevalence for both groups was negative, but the Coombs and Wright tests score was 33 (20.6%) in pregnant women and it was 27 (16.9%) in women who experienced spontaneous abortion. No meaningful relationship was observed between spontaneous abortion and brucellosis (p= 0.39). Conclusion: Even though the present study did not find a meaningful relationship between spontaneous abortion and brucellosis (p=0.39), high brucella seroprevalence rates between both groups of women indicated that screening tests should be considered before gestation as an appropriate therapeutic strategy.

Retraction Note: Down-regulation of miR-133a and miR-539 are associated with unfavorable prognosis in patients suffering from osteosarcoma.

[This retracts the article DOI: 10.1186/s12935-015-0237-6.].

Genetic modification of bone-marrow mesenchymal stem cells and hematopoietic cells with human coagulation factor IX-expressing plasmids.

Ex-vivo gene therapy of hemophilias requires suitable bioreactors for secretion of hFIX into the circulation and stem cells hold great potentials in this regard. Viral vectors are widely manipulated and used to transfer hFIX gene into stem cells. However, little attention has been paid to the manipulation of hFIX transgene itself. Concurrently, the efficacy of such a therapeutic approach depends on determination of which vectors give maximal transgene expression. With this in mind, TF-1 (primary hematopoietic lineage) and rat-bone marrow mesenchymal stem cells (BMSCs) were transfected with five hFIX-expressing plasmids containing different combinations of two human ß-globin (hBG) introns inside the hFIX-cDNA and Kozak element and hFIX expression was evaluated by different methods. In BMSCs and TF-1 cells, the highest hFIX level was obtained from the intron-less and hBG intron-I,II containing plasmids respectively. The highest hFIX activity was obtained from the cells that carrying the hBG intron-I,II containing plasmids. BMSCs were able to produce higher hFIX by 1.4 to 4.7-fold increase with activity by 2.4 to 4.4-fold increase compared to TF-1 cells transfected with the same constructs. BMSCs and TF-1 cells could be effectively bioengineered without the use of viral vectors and hFIX minigene containing hBG introns could represent a particular interest in stem cell-based gene therapy of hemophilias.

An optimised mouse model of chronic pancreatitis with a combination of ethanol and cerulein.

INTRODUCTION: Chronic pancreatitis (CP) is an intractable and multi-factorial disorder. Developing appropriate animal models is an essential step in pancreatitis research, and the best ones are those which mimic the human disorder both aetiologically and pathophysiologically. The current study presents an optimised protocol for creating a murine model of CP, which mimics the initial steps of chronic pancreatitis in alcohol chronic pancreatitis and compares it with two other mouse models treated with cerulein or ethanol alone. MATERIAL AND METHODS: Thirty-two male C57BL/6 mice were randomly selected, divided into four groups, and treated intraperitoneally with saline (10 ml/kg, control group), ethanol (3 g/kg; 30% v/v), cerulein (50 µg/kg), or ethanol + cerulein, for six weeks. Histopathological and immunohistochemical assays for chronic pancreatitis index along with real-time PCR assessments for mRNA levels of inflammatory cytokines and fibrogenic markers were conducted to verify the CP induction. RESULTS: The results indicated that CP index (CPI) was significantly increased in ethanol-cerulein mice compared to the saline, ethanol, and cerulein groups (p < 0.001). Interleukin 1ß (IL-1ß), tumor necrosis factor α (TNF-α), transforming growth factor ß (TGF-ß), α-smooth muscle actin (α-SMA), and myeloperoxidase activity were also significantly greater in both cerulein and ethanol-cerulein groups than in the saline treated animals (p < 0.001). Immunohistochemical analysis revealed enhanced expression of TGF-ß and α-SMA in ethanol-cerulein mice compared to the saline group. CONCLUSIONS: Intraperitoneal (IP) injections of ethanol and cerulein could successfully induce CP in mice. IP injections of ethanol provide higher reproducibility compared to ethanol feeding. The model is simple, non-invasive, reproducible, and time-saving. Since the protocol mimics the initial phases of CP development in alcoholics, it can be used for investigating basic mechanisms and testing new therapies.

Diagnostic procedures for improving of the KIT (CD117) expressed allele burden for the liver metastases from uterus mast cell tumors: prognostic value of the metastatic pattern and tumor biology.

The activating KIT marker plays a central role in the pathogenesis, diagnosis, and targeted treatment of systemic mastocytosis (SM). Recent studies have identified the KIT (CD117) as a marker that distinguishes nonneoplastic from neoplastic mast cells in human systemic mastocytosis. In this study, we conclude that immunohistopathology assays for KIT staining pattern are useful complimentary tools for diagnosis and evaluation of prognosis in uterus mast cell tumor (MCT) metastasis to the liver in 10 patients. Uterine and hepatic cytology revealed mast cell neoplasia, which was confirmed as visceral mast cell tumor on postmortem examination. Histological changes of densely packed, poorly differentiated neoplastic mast cells, sheets of neoplastic round to pleomorphic cells that formed nonencapsulated nodules, high mitotic figures, necrosis, and fibrosis were found. In addition, eosinophils were scattered among the mast cells at the periphery of the nodules. These findings indicate tumors of high-grade malignancy with infiltrative cells resembling the uterus MCT in the intraparenchymal and periparenchymal areas of the liver. Immunohistochemically, tumors were positive for KIT. The histopathologic features coupled with the KIT immunoreactivity led to diagnosis of high-grade uterus MCTs. Taken together, these findings suggest that CD117 may play a critical role in early uterus MCT development and may be a stimulatory factor in grade 3 MCT. Therefore, the result has supported our hypothesis that there was an increased opportunity to observe a higher CD117 staining pattern in high-grade MCTs.

Diagnostic investigations of canine prostatitis incidence together with benign prostate hyperplasia, prostate malignancies, and biochemical recurrence in high-risk prostate cancer as a model for human study.

The aim of this study was to evaluate the prevalence of acute and chronic inflammation, benign prostatic hyperplasia (BPH), and cancer of the prostate glands in the canine as a human model in prostate disorders. The study was carried out on 12 cases of different male dogs of terrier (50%), German shepherd (25%) breeds, and Greden (25%), and the age of the dogs ranged from 6 to 13 years (average age 7.8 ± 3.6). The bodyweight ranged from 3.6 to 7.9 kg. Signalment, clinical signs, and diagnostic tools such as ultrasonography, urinary cytology, and histopathology are presented. Dysuria was the most common clinical sign in this study and occurred in 10/12 canine (83.3%) included. Other clinical signs included lameness (5/12 canine, 41.6%) and constipation (3/12 canine, 25%). The range of duration of clinical signs was 5 days to 7 months. Moreover, in the present study, the urinary biochemical markers of different prostate lesions include blood, protein, and glucose and were detected in 11/12 cases (91.6%), 5/12 cases (41.6%), and 2/12 cases (16.6%), respectively. Taken together, sonographic data were classified into four groups based on histological diagnosis. In 7/12 cases (58.4%), the prostate appeared to have BPH lesions, and the remaining lesions included inflammation (3/12 cases, 25%), abscess (1 case, 8.3%), and adenocarcinoma (1 case, 8/3%) on ultrasound. In all cases, prostate tissue had an irregular echotexture. None of the dogs had sonographic evidence of sublumbar lymph node enlargement. Histopathologically, we looked at the prevalence of inflammation (33.3% chronic and 8.3% acute) and BPH (58.4%) in dogs of different ages and breeds, and also, we observed chronic inflammation in >20% of dogs, which was about 25% in 3 cases of the 12 cases referred. More chronic inflammation was associated with more BPH. The majority of the asymptomatic inflammation that is detected in the prostate is classified as chronic inflammation (i.e., as evidenced by the presence of monocytic and/or lymphoplasmacytic inflammatory cell infiltrates); however, acute inflammation is also observed to a lesser degree. Acute inflammation, as is typically evidenced by the infiltration of neutrophils, is classically an indicator of an infectious process. Finally, the patients included seven castrated, four castrated together with antibiotic therapy, and one castrated together with chemotherapy intact male dogs, which were treated with the mentioned cases. In conclusion, chronic prostatic inflammation could be a central mechanism in BPH progression, but the pathological features of tissue inflammation were different between BPH and prostate cancer (PCa). Nevertheless, the histological examination of prostate biopsies remains the only way to diagnose prostatic disorders.

Down-regulation of miR-133a and miR-539 are associated with unfavorable prognosis in patients suffering from osteosarcoma.

BACKGROUND: MicroRNAs (miRNAs) play key roles in cancer development and progression. The purpose of the present study was to determine the expression levels of miR-133a and miR-539 in osteosarcoma patients and to further investigate the clinicopathological, and prognostic value of these miRNAs. METHODS: The expression levels of miR-133a and miR-539 were determined by qRT-PCR. Associations between miRNAs expressions and various clinicopathological characteristics were analyzed. Survival rate was determined with Kaplan-Meier and statistically analyzed with the log-rank method between groups. Survival data were evaluated through multivariate Cox regression analysis. RESULTS: Our findings revealed that the miR-133a expression was significantly decreased in clinical osteosarcoma tissues compared to adjacent normal bone tissues. The expression level of miR-539 was decreased in clinical osteosarcoma tissues as compared to those adjacent normal tissues. Low expressions of miR-133a and miR-539 were significantly association with advanced TNM stage (P = 0.002; P = 0.001), and metastasis or recurrence (P = 0.001; P = 0.01). Furthermore, Kaplan-Meier survival analysis and log-rank test showed that the low expressions of miR-133a and miR-539 were correlated with the reduced overall survival of osteosarcoma patients. Multivariate Cox proportional hazards model showed that decreased expressions of miR-133a and miR-539 (P = 0.007; P = 0.02), TNM stage (P = 0.001; P = 0.002), and metastasis or recurrence (P = 0.005; P = 0.026) were independent prognostic markers of overall survival of patients. CONCLUSION: These results suggest that decreased miR-133a and miR-539 expressions may participate in the progression of osteosarcoma. Together, these results showed that miR-133a and miR-539 may have their role in both progression and prognosis of osteosarcoma.

A comprehensive review of pesticides and the immune dysregulation: mechanisms, evidence and consequences.

Nowadays, in many communities, there is a growing concern about possible adverse effects of pesticides on human health. Reports indicate that during environmental or occupational exposure, pesticides can exert some intense adverse effects on human health through transient or permanent alteration of the immune system. There is evidence on the relation between pesticide-induced immune alteration and prevalence of diseases associated with alterations of the immune response. In the present study, direct immunotoxicity, endocrine disruption and antigenicity have been introduced as the main mechanisms working with pesticides-induced immune dysregulation. Moreover, the evidence on the relationship between pesticide exposure, dysregulation of the immune system and predisposition to different types of psychiatric disorders, cancers, allergies, autoimmune and infectious diseases are criticized.

Stromal vascular fraction combined with silicone rubber chamber improves sciatic nerve regeneration in diabetes.

PURPOSE: To study the effects of transplantation of characterized uncultured stromal vascular fraction (SVF) on sciatic nerve regeneration. METHODS: A 10-mm sciatic nerve defect was bridged using a silicone conduit filled with SVF. In control group, silicone conduit was filled with phosphate-buffered saline alone. In sham-operated group, the sciatic nerve was only exposed and manipulated. The regenerated nerve fibers were studied 8 and 12 weeks after surgery. RESULTS: Behavioral and functional studies confirmed faster recovery of regenerated axons in SVF transplanted animals than in control group (p<0.05). Gastrocnemius muscle mass in SVF transplanted animal was found to be significantly more than that in control group. Morphometric indices of the regenerated fibers showed the number and diameter of the myelinated fibers to be significantly higher in SVF transplanted animals than in control group. In immunohistochemistry, the location of reactions to S- 100 in SVF transplanted animals was clearly more positive than that in control group. CONCLUSION: SVF transplantation combined with silicone conduit could be considered as a readily accessible source of stromal cells that improves functional recovery of sciatic nerve. It may have clinical implications for the surgical management of acute diabetic patients after facial nerve transection.

Improving the diagnosis, treatment, and biology patterns of feline mammary intraepithelial lesions: a potential model for human breast masses with evidence from epidemiologic and cytohistopathologic studies.

In this study, the frequency of different types of mammary masses and their relationship with cytohistopathologic changes was investigated and data on history, macroscopic description, clinical examination and treatment were collected. To determine the prevalence and types of cytohistopathologic changes, mammary glands from 12 female cats were evaluated. The mean age of cats at the time of diagnosis was 11.5 ± 1.9 years (range 4-14 years), the mean gross size of the masses was 3.1 ± 2.4 cm, 4/12 (33.3 %) masses were ≤3.0 cm in diameter, and the maximum diameter of the largest mass had a median of 5 cm, with a range of diameter of 6 × 5 × 4 cm. Moreover, the preferential localization of mammary masses was the abdominal lobes (%50) and thoracic lobes (%33.3), and inguinal lobes (%16.7 of cases). Furthermore, two cases of the inguinal masses affected the caudo-inguinal lobe, six cases caudo-abdominal lobe, and thoracic masses were found in four cases. Eventually, six cases (%50) of masses were found in the right mammary lobes and six cases (%50) in the left mammary lobes. The majority of the masses revealed elastic (%50 of cases), hard (%25 of cases), or soft (%25 of cases) consistency. In the present study, according to the criteria of the veterinary and the medical WHO classification system, of the 12 cats with the cytohistopathological features of six (50 %) cases qualified abscess, 3 (25 %) cases as cystic hyperplasia and 3 (25 %) cases were called situ carcinoma. Whereas, all hyperplastic lesions (case nos. 7-9 and ranging in size from, 1 to >4 cm(3)) and carcinomas in situ lesions (case nos. 10-12 and ranging in size from, 1 to >3 cm(3)) were found incidentally upon routine cytohistology. Other lesions were observed grossly and removed either at surgery (case nos. 1-6). Finally, the cats were treated with unilateral lumpectomy (3 cases) and also, nine (75 %) cases had subsequent drainage, 3 (25 %) of which showed cystic hyperplasia and 6 (50 %) showed abscess on subsequent histopathological evaluation. Therefore, a correct diagnosis must be established quickly, and treatment must be instituted rapidly when alteration is noted in the mammary glands.

Clinico-histopathologic and outcome features of cutaneous infundibular keratinizing acanthoma: a case report and literature review.

The infundibular keratinizing acanthoma (IKA) is a rare epithelial benign keratin-containing neoplasm of hair follicles. The purpose of this study was to evaluate the defining histopathologic architecture of IKA. A typical IKA consisted of a keratin-filled crypt in the dermis and subcutis that opened to the skin surface. Most of this tumor occurred on the back, neck, head, and the shoulders. Microscopically, the dermal nodules were focally contiguous in both the dermis and subcutis. Furthermore, most histological lesions are consistent with a simple or multiloculated cyst filled with keratin and lined by a wall of stratified squamous epithelium; keratin appears as a concentric lamellar mass, with a keratotic pearly aspect. Histological examination of the cutaneous lesions revealed that the growths were comprised of IKA. IKA of man and dog were compared, and it was concluded that although they are similar in many respects, they are not identical entities. To the best of our knowledge, this is the first report on the prevalence of IKAs among the population of owned dogs in Iran.

The court's judgment on the true identity of the author responsible for identity fraud and the consequent batch retraction of Iranian's papers in the journal of diagnostic pathology.

This letter concerns retracted papers published in the Journal of Diagnostic Pathology, where my name was misused as the author or corresponding author without my permission or knowledge. Considering that all misconducts were directed by an author during initial manuscripts' submissions, I opened a case in Iran's Cyber Police (FATA) to unravel the true identity of the submitting author. After Cyber Police's report revealed the true identity of the submitting author, the court started a thorough investigation and finally convicted the submitting author for identity fraud and data forgery through creating and using fake email addresses.

Biomarkers for Assessment of Human Immunodeficiency Virus and its Co-Infection with Hepatitis B and Hepatitis C Viruses: A Comprehensive Review.

Recently, prevalence of hepatitis B virus (HBV), and hepatitis C virus (HCV) co-infection with Human immunodeficiency virus (HIV), has dramatically increased worldwide due to their shared routes of transmission. Compared to the sporadic infection with HIV, HBV, and HCV, concurrent infection with these agents increases the complications of these viruses. Furthermore, co-infection may also alter the therapeutic strategies against HIV. Accordingly, choosing appropriate biomarkers to detect these co-infections is one of the main concerns in the field of diagnostic pathology. Up to now, several markers have been introduced for the simultaneous diagnosis of HIV, HBV, and HCV. In this regard, serum adenosine deaminase activity (ADA), FibroTests, AST-to-Platelet Ratio Index (APRI), Fibrosis-4, Hyaluronic acid, and micro ribonucleic acids (MiR) have been investigated as potential biomarkers for diagnosis of HIV-HCV/HBV co-infections. This review summarizes diagnostic values of the current and emerging biomarkers in HIV patients concurrently infected with HBV and HCV.

Designing a Strategy for pH Control to Improve CHO Cell Productivity in Bioreactor.

BACKGROUND: Drastic pH drop is a common consequence of scaling up a mammalian cell culture process, where it may affect the final performance of cell culture. Although CO2 sparging and base addition are used as common approaches for pH control, these strategies are not necessarily successful in large scale bioreactors due to their effect on osmolality and cell viability. Accordingly, a series of experiments were conducted using an IgG1 producing Chinese Hamster Ovary (CHO-S) cell culture in 30 L bioreactor to assess the efficiency of an alternative strategy in controlling culture pH. METHODS: Factors inducing partial pressure of CO2 and lactate accumulation (as the main factors altering culture pH) were assessed by Plackett-Burman design to identify the significant ones. As culture pH directly influences process productivity, protein titer was measured as the response variable. Subsequently, Central Composite Design (CCD) was employed to obtain a model for product titer prediction as a function of individual and interaction effects of significant variables. RESULTS: The results indicated that the major factor affecting pH is non-efficient CO2 removal. CO2 accumulation was found to be affected by an interaction between agitation speed and overlay air flow rate. Accordingly, after increasing the agitation speed and headspace aeration, the culture pH was successfully maintained in the range of 6.95-7.1, resulting in 51% increase in final product titer. Similar results were obtained during 250 L scale bioreactor culture, indicating the scalability of the approach. CONCLUSION: The obtained results showed that pH fluctuations could be effectively controlled by optimizing CO2 stripping.