The effects of Fasciola hepatica recombinant proteins (peroxiredoxin and cathepsin L1) on Crohn's disease experimental model.

The immunomodulatory potential of the excretory-secretory (E/S) proteins of the helminths has been shown in previous investigations. This study evaluated the effects of the recombinants and excretory-secretory proteins of the Fasciola hepatica on induced colitis in Balb/c mice. The F. hepatica Recombinant proteins, Cathepsin L1 and Peroxiredoxin, and E/S proteins were intraperitoneally injected into the three mice groups as the case groups, while the control groups received PBS. Colitis was induced in mice by intraluminal administration of the 2, 4, 6-Trinitrobenzenesulfonic acid solution (TNBS). After 8 h, the case groups received the second dosage of the treatments, and it was repeated 24 h later. The immunological, pathological, and macroscopic changes were evaluated 3 days after colitis induction. The macroscopic evaluation revealed significantly lower inflammatory scores in the mice treated with recombinant Peroxiredoxin (rPRX) and recombinant Cathepsin L1 (rCL1). Despite the macroscopic observation, the pathological finding was insignificant between the groups. IFN-γ secretion was significantly lower in splenocytes of the groups that received rPRX, rCL1, and E/S than the controls. IL-10 showed significantly higher levels in groups treated with rPRX and rCL1 than controls, whereas the level of IL-4 was not statistically significant. Excretory-secretory proteins of the F. hepatica showed immunomodulatory potency and the main effects observed in this study were through the reduction of inflammatory cytokine and inflammation manifestation as well as induction of anti-inflammatory cytokines.

TLRs expression in canine mammary gland neoplasms: a pathological and molecular study.

TLRs are a class of PRRs that play a vital role in innate immunity. TLRs are expressed on immune cells and mammary epithelial cells. They can promote tumor growth, angiogenesis, invasion, and viability signaling. The current study aimed to test the correlation between histologic types and grades of neoplasms and TLRs gene expression levels. Twenty-one tissue samples of canine mammary neoplasms were stained with H&E. Then, it evaluated histologic type and grade according to the methods of Goldschmidt et al. and Peña, respectively. We established real-time PCR quantification assays to measure the mRNA abundances of TLRs in normal and neoplastic mammary glands. Profile pattern of TLR 1, 2, 3, 4, 5, 6, and 9 genes expression in canine mammary glands performed in 21 samples of mammary gland neoplasms and three non-neoplastic mammary gland samples from normal dogs. TLR 3, 4, and 9 mRNA overexpression were detected. In addition, tubulopapillary carcinoma grade II, SCC grade III, and carcinoma mixed type grade II demonstrated the highest relative TLR-3, and 9 mRNA expression levels. Complex carcinoma grade I, ductal carcinoma grade II, and anaplastic carcinoma grade II showed the highest relative TLR4 mRNA expression level. Although histopathological characteristics of tumors, including histologic type, grade, and inflammation, influenced TLRs mRNA expression level, such correlation was insignificant (P > 0.05).

Molecular Characterization of Fasciola spp. from Some Parts of Iran.

BACKGROUND: Identification of liver flukes, Fasciola hepatica, and Fasciola gigantica by morphometric parameters is not always reliable due to the overlapping measurements. This study aimed to characterize the liver flukes of animals from different parts of Iran by the genetic markers, ITS1, and COXI. METHODS: We collected flukes from infected livestock in six provinces of Iran from Sep to Nov 2016. The flukes were identified by amplification of a 680 bp sequence of ITS1 locus followed by a restriction fragment polymorphism (RFLP) assay. The genetic diversity among isolates was evaluated by amplification and sequencing of a 493 bp fragment of the COXI gene. RESULTS: We obtained 38 specimens from Khuzestan, 22 from Tehran, 10 from Isfahan, 10 from Mazandaran, 4 from Kurdistan, and 3 from Ardabil provinces. PCR-RFLP analysis revealed two patterns, representing F. hepatica, and F. gigantica. Fifty specimens from cattle and sheep exhibited F. hepatica pattern and 37 from the cattle, sheep, buffalo, and goat that of F. gigantica. The phylogeny based on COXI revealed two distinct clades separating F. hepatica from F. gigantica. In our phylogeny, the Iranian F. gigantica isolates showed a distinct separation from the African flukes, while grouped with the East Asia specimens demonstrating a common ancestor. The F. hepatica isolates clustered with the flukes from different parts of the world, including East Asia, Europe, and South America. CONCLUSION: The present study revealed a substantial genetic difference between F. gigantica populations of Asia and Africa, while F. hepatica isolates from different parts of the world shared high similarities.

Esophageal-Crop Capillariasis and Proventriculus Ventriculus Hystrichisiasis in a Migratory Duck (Anas crecca).

BACKGROUND: The aim of the present study was to study pathological and parasitological characteristics of simultaneous capillariasis and hystrichiasis in a diseased Anas crecca captured in Mazandaran Province of Iran on Oct 2016. METHODS: Parasitology and histopathology techniques were used to detect the parasites of the carcass of the captive common teal. RESULTS: Macroscopically, severe chronic inflammatory reactions and nodular or granuloma formation and irregular thickening of the affected wall of the esophagus, crop, proventrculus and ventriculus were observed. Microscopically, apart from the chronic inflammatory reactions and granuloma formation, in the paraffin sections stained with Harris Hematoxyline and Eosin technique, the characteristics of the mature female and male Capillaria spp. and their barrel-shaped operculated embryonated eggs were found within the tunnels burrowed by the nematode in the hyperplastic stratified squamous epithelium of the inflamed crop and distal portion of the esophagus. The mature female Hystrichis spp. containing oval-shaped, embryonated non-operculated eggs and male parasite were found within the labyrinthus spaces formed in the submucosa granulomas of the ventriculus and proventriculus. CONCLUSION: Based on the parasitological and pathological studies, the species of the nematode parasites were identified as Capillaria (Thominx) contorta and Hystrichis. tricolor.

Correction to: Blocking of opioid receptors in experimental formaline-inactivated respiratory syncytial virus (FI-RSV) immunopathogenesis: from beneficial to harmful impacts.

In the original publication, seventh author's name was incorrectly published as 'Maryam Golaram'.

Blocking of opioid receptors in experimental formaline-inactivated respiratory syncytial virus (FI-RSV) immunopathogenesis: from beneficial to harmful impacts.

Opioid system plays a significant role in pathophysiological processes, such as immune response and impacts on disease severity. Here, we investigated the effect of opioid system on the immunopathogenesis of respiratory syncytial virus (RSV) vaccine (FI-RSV)-mediated illness in a widely used mouse model. Female Balb/c mice were immunized at days 0 and 21 with FI-RSV (2 × 106 pfu, i.m.) and challenged with RSV-A2 (3 × 106 pfu, i.n.) at day 42. Nalmefene as a universal opioid receptors blocker administered at a dose of 1 mg/kg in combination with FI-RSV (FI-RSV + NL), and daily after live virus challenge (RSV + NL). Mice were sacrificed at day 5 after challenge and bronchoalveolar lavage (BAL) fluid and lungs were harvested to measure airway immune cells influx, T lymphocyte subtypes, cytokines/chemokines secretion, lung histopathology, and viral load. Administration of nalmefene in combination with FI-RSV (FI-RSV + NL-RSV) resulted in the reduction of the immune cells infiltration to the BAL fluid, the ratio of CD4/CD8 T lymphocyte, the level of IL-5, IL-10, MIP-1α, lung pathology, and restored weight loss after RSV infection. Blocking of opioid receptors during RSV infection in vaccinated mice (FI-RSV-RSV + NL) had no significant effects on RSV immunopathogenesis. Moreover, administration of nalmefene in combination with FI-RSV and blocking opioid receptors during RSV infection (FI-RSV + NL-RSV + NL) resulted in an increased influx of the immune cells to the BAL fluid, increases the level of IFN-γ, lung pathology, and weight loss in compared to control condition. Although nalmefene administration within FI-RSV vaccine decreases vaccine-enhanced infection during subsequent exposure to the virus, opioid receptor blocking during RSV infection aggravates the host inflammatory response to RSV infection. Thus, caution is required due to beneficial/harmful functions of opioid systems while targeting as potentially therapies.

In Vitro Assay of Paecilomyces lilacinus Biocontrol Effects on Fasciola hepatica Eggs Illustrated in Scanning Electron Micrographs.

BACKGROUND: Fascioliasis is a zoonotic disease caused by the liver fluke Fasciola hepatica. Drug resistance, high costs of treatment and economic losses in meat production have emerged the need of alternative control measures into consideration. The aim of this study was to evaluate the in vitro ovicidal activity of Paecilomyces lilacinus fungus on F. hepatica eggs. METHODS: P. lilacinus isolated from the soil of natural environment was challenged on F. hepatica eggs to observe the bio control effect of nematophagous fungi on trematode helminth eggs. The study was conducted in Tehran University of Medical Sciences, in 2015. Within 21 d of experiment, destructive effects exhibited on the eggshells were investigated using optical and Scanning Electron Microscopy. RESULTS: The effective role of P. lilacinus on damaging the eggs of F. hepatica was noticed. CONCLUSION: This finding is promising for advantageous use of nematophagus fungi as a natural constituent in hyper endemic areas for certain helminthic infections like fascioliasis with diverse kinds of herbivores as egg passer hosts.

Preliminary investigation of the effects of topical mixture of Lawsonia inermis L. and Ricinus communis L. leaves extract in treatment of osteoarthritis using MIA model in rats.

BACKGROUND: Many plants have been introduced in Iranian traditional medicine for treatment of different joint problems including knee pain. Topical application of the mixture of Lawsonia inermis L. leaves (Henna) with aqueous extract of Ricinus communis L. leaves have been mentioned to have significant effects on reducing knee pain. The present study was designed to evaluate the analgesic and anti-inflammatory effects of the mixture of these two herbs in male rats. METHODS: We induced knee osteoarthritis as a model of chronic pain by intra-articular injection of mono sodium iodoacetate (MIA). Mechanical allodynia, hotplate latency test, spontaneous movements and gait analysis were used for the evaluation of analgesic activity. Anti-inflammatory activity was evaluated by measuring the diameter and the volume of the injected paw compared to contralateral paw. These tests were monitored at days 1, 3, 7, 14 and 21 of MIA administration. Histopathological evaluations were also used to assess the efficacy of the treatment on inflammation and lesions in knee tissue. In all tests, diclofenac topical gel was used as a positive control. The herbal extracts, their mixture, and vehicle or diclofenac gel were administered daily for 14 days by topical route. RESULTS: The mixture of these two extracts significantly reduced the knee joint width and volume of the injected paws and also improved foot prints in gait analysis after 3 days of MIA injection. Analysis of mechanical allodynia (after 21 days), hotplate latency test (after 10 days), spontaneous movements (after 7 days) and in positive control group (after 3 days in all tests and in mechanical allodynia after 14 days) compared to the vehicle group, showed significant effects. Topical usage of the selected formulation made significant histopathological changes on the knee of the rats. Compared to the vehicle group, the tests and diclofenac groups showed less reactions characterized by negligible edema and a few scattered inflammatory lymphoid cells. CONCLUSION: The present findings showed that the present formulation not only was able to mitigate pain and inflammation in the paws but also made significant histopathological changes on the knee of the rats. Further studies are necessary to confirm the effect of the formulation.

Metastatic ovarian papillary cystadenocarcinoma to the small intestine serous surface: report of a case of high-grade histopathologic malignancy.

Ovarian cystadenocarcinoma is characterized by marked heterogeneity and may be composed of an admixture of histologic growth patterns, including acinar, papillary and solid. In the present study, a case of isolated small intestine metastasis of ovarian papillary cystadenocarcinoma was reported. A 7-year-old female mixed-breed dog presented with a mass in the left upper quadrant with progressive enlargement of the abdomen, periodic bloody discharge from the vulva and incontinence. The tumor was histologically characterized by the presence of cysts and proliferation of papillae, both lined by single- or multi-layered pleomorphic epithelial cells. Furthermore, the mass was composed by intense cellular and nuclear pleomorphism and numerous mitotic figures. These findings indicate a tumor of high-grade malignancy with infiterative tumor cells resembling the papillary ovarian tumor in the serosal surface of the small intestine along with an intact serosa. Immunohistochemically, tumor was positive for CK7 and negative immunoreactivity for CK20. The histopathologic features coupled with the CK7 immunoreactivity led to a diagnosis of high grade ovarian papillary cystadenocarcinoma. To the best of our knowledge, this is the first case of small intestine serousal surface metastasis from ovarian papillary cystadenocarcinoma.

Cervical type AB thymoma (Mixed) tumour diagnosis in a mynah as a model to study human: clinicohistological, immunohistochemical and cytohistopathological study.

Thymoma is a primary mediastinal neoplasm arising from or exhibiting differentiation towards thymic epithelial cells, typically with the presence of non-neoplastic lymphocytes. A 13-year-old male Mynah bird (acridotheres tristis) was presented for evaluation of a 2.3 × 1.5 × 1.0 cm mass in the left ventrolateral cervical region. The clinical signs, radiology, cytohistopathology and immunohistochimy findings related to the thymoma are presented. These findings indicated that the tumor was a type AB thymoma according to the World Health Organization (WHO) and veterinary classification. Thymomas are rarely reported in avian species and this is the first report in a Mynah bird. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1159525819982779.

The Oncolytic Effect of Respiratory Syncytial Virus (RSV) in Human Skin Cancer Cell Line, A431.

BACKGROUND: Oncolytic viruses have become of noticeable interest as a novel biological approach for selectively infecting cancer cells and triggering apoptosis in a number of malignant cells. Many researches are devoted to characterize more viruses with oncolytic properties. OBJECTIVES: Evidences on the oncolytic feature of respiratory syncytial virus (RSV) are conflicting; therefore, this study was designed to elucidate the possible role of RSV on the modulation of cell growth and apoptosis in the skin cancer cells. MATERIALS AND METHODS: Plaque assay was used to determine RSV titers. The cytotoxic effect of RSV in A431 (skin carcinoma cell line) was determined using MTT assay. The detection of apoptosis was performed via Annexin-V-FITC staining method and analyzed with flow cytometry. RESULTS: The results indicated that A431 cell growth was inhibited following infection by RSV in a dose- and time-dependent manner. The most growth inhibitory effect of RSV was occurred at the MOI of 3, and 48 hour after infection. The inhibitory effect of RSV on the cell growth was accompanied by the induction of apoptosis in the skin cancer cells. The percentages of early and late apoptotic cells were increased following exposure to RSV in a concentration- and time-dependent manner. CONCLUSIONS: This study delineated the beneficial role of RSV for growth regulation of skin cancer cells and highlighted the involvement of RSV in the induction of apoptosis in A431 cells. These findings might conduct evidence into the oncolytic properties of RSV in the skin cancer. Further studies are required to indicate intracellular targets for RSV-induced apoptosis in skin cancer cells.

Post challenging serum cytokine profile (Th1 & Th2) in the vaccinated mice (Balb/C) with a new formulation of Leishmania major antigen.

OBJECTIVE: The aim of this study was to carry out experiments further to our previous new formulation to modify the Leishmania major antigen that had satisfactory results previously. METHODS: In this study we made a preliminary, new vaccine with the same methodology and selected two injection doses (100&200 µg/o.1 mL), three injection Groups: Leishmania plus BCG (LB), Leishmania plus new adjuvant (Teucrium Polium) [LT], Leishmania plus BCG and Teucrium Polium (LBT), and one susceptible mouse Group (Balb/c) and measure two types of cytokines: Th1 (IFN-γ, IL-12) and Th2 (IL-4, IL-10) We prepared crude antigen combinations by five different methods using antigens from L. major parasites. Phase I was done in the animal model. In our study, Leishmania antigen was examined both with BCG and the new adjuvant (TP) in three Groups in two injection doses (100.200 µg/1 mL) and Balb/c mice. RESULTS: Our results showed that in three injection Groups (LB, LT and LBT) that received each or both BCG and TP as adjutant with injection doses of 100 and 200 µg/1 mL with two booster doses: the LBT Group had the lowest IFNγ and highest IL-12 value, LT and LB Groups have equal IL-12, but LB have more IFNγ and IL-10 but less than IL-4 in the LT Group. CONCLUSION: In this study, the LBT Group has statistical differences regarding IL-12 and IL-10 from the other Groups.

In vivo electroporation enhances immunogenicity and protection against influenza A virus challenge of an M2e-HSP70c DNA vaccine.

There is a growing concern regarding continuous risk of emerging a new influenza pandemic. It is highlighted the need for novel vaccination techniques that quickly and effectively employed to respond to such threats. Although, DNA vaccine is a simple and effective approach to induce antigen specific immune responses, their potency requires further improvement. DNA vaccine encoding conserved antigen of influenza virus could provide protection in various animal models. Therefore, we constructed a plasmid vector encoding M2e-HSP70c sequences, pcDNA/MHc, as a candidate for universal influenza vaccine. The expression of newly constructed vectors was verified by transient transfection of mammalian cells (HEK293T cell line) and western blot analysis using commercial antibodies. Mice were injected subcutaneously (s.c.) by the help of electroporation (IEP) in the footpad area and boosted without IEP with 100 µg of constructed vector. Furthermore, the potency of this construct to provoke humoral immune responses and its protectivity against lethal dose of viral challenge were evaluated. Based on our study, the fusion construct was immunogenic in mice and was able to confer both protection against lethal challenge of H1N1 virus and reduce viral load in lung homogenates of the infected mice.

Endoscopic treatment of vesicoureteral reflux using calcium hydroxyl apatite in dogs.

BACKGROUND: Injection of biomaterial to suburetral region, using minimally invasive procedure, has become an interesting topic for urologists to treat vesicoureteral reflux. The objective of this study was to evaluate the feasibility of injecting newly introduced calcium hydroxyl apatite to suburetral region, for treating an experimentally induced vesicoureteral reflux in dogs. FINDINGS: Bilateral vesicoureteral refluxed (VUR) mixed breed dogs (n = 12; 10-15 kg live weight, 3-6 months of age) were selected for this study. The presence and grade of the reflux were determined using cystography. Accordingly, 6 dogs displayed grade 1 & 2 and the other 6 showed grade 3 & 4 bilateral VUR. Every single dog, with bilateral VUR, underwent endoscopic treatment and received an injection of calcium hydroxyl apatite (an Iranian made product) into the left (treated side) and an injection of the similar volume of normal saline in to the right (control side) subureteric space. One week, 3 and 6 months after treatment, cystography was performed. On each occasion, 4 dogs were euthanized by gas inhalation and biopsy samples were collected for histopathological study from ureter, bladder, kidney, lung and spleen in order to investigate the biomaterial migration into different organs. Data were analyzed using Chi-squared test. In control sides, radiographs confirmed the same grade of VUR, found at the initiation of the study. VUR was resolved in 100% (6/6) of Grade 1 & 2 and 83.33% (5/6) of Grade 3 & 4 in treated side. Therefore, the total success rate of this study was 91.67% (11/12). Macroscopic examination of the vesicouretral region of the treated side revealed a firm and consistent biomaterial mass at the site of injection. Histological findings confirmed inflammation at treated side. In contrast, there was no tissue reaction on control side. There was no evidence for biomaterial migration in macroscopic and microscopic observations in this study. CONCLUSION: In the present study, a new biocompatible material produced a firm, consist and sustainable biomaterial mass in the suburetral region for treating vesicouretral reflux without any evidence of biomaterial migration.

Single-layer versus double-layer laparoscopic intracorporeally sutured gastrointestinal anastomoses in the canine model.

OBJECTIVE: The objective of this study was to compare the gross and histopathologic changes following 1- versus 2-layer hand-sewn suture techniques in laparoscopic gastrointestinal anastomosis in dogs. METHODS: This was an experimental prospective study of 16 healthy mixed breed male and female dogs. Animals were randomly divided into 2 groups. Two-layer side-to-side hand-sewn laparoscopic gastrojejunostomies were performed in group A, so that simple interrupted sutures were placed in the outer layer and simple continuous suture was used in the inner layer. The 1-layer simple continuous anastomosis between the stomach and jejunum was done in group B precisely. Specimen were collected from the sites of anastomosis, and H&E statining was performed for light microscopic studies. RESULTS: All animals survived the surgery. There was no gross inflammation, ischemia, apparent granulation tissue, abscess or fistula formation, leakage or stricture formation, and all sites of anastomosis were patent. Several adhesion formations were found in the abdomen with the higher incidence in the control group. Mean scores of leukocyte infiltration and granulation tissue formation at the sites of anastomosis were statistically insignificant between groups (P>0.05). CONCLUSIONS: Gross and histopathologic findings revealed that hand-sewn laparoscopic gastrointestinal anastomosis with the 1-layer suture technique is comparable to the 2-layer suture technique.

Cochlosoma infection in a turkey in Iran.

Cochlosoma sp. infection was identified in a single case among 60 stunted diarrheic native turkey poults, Meleagris galopavo. A large number of the flagellated parasites was found free or within the intervillous spaces of the jejunum, ileum and cecum. Moderate enteritis was associated with the parasites. In TEM studies of the parasagittal sections of the parasite, a prominent ventral sucker like disc and flagella emerging from an opening on the ventrodorsal surface of the pyriform uninuclear parasite were found. The morphological characteristics of this protozoan match with those described for Cochlosoma anatis. The parasite could be considered as an intestinal pathogenic protozoan causing stunting and diarrhea in turkeys in Iran.

Neutrophil function (innate immunity) during Ramadan.

BACKGROUND: Fasting during the month of Ramadan is one of the essential religious practices of Muslims. The aim of this study was to evaluate opsonisation, phagocytosis, and nitroblue tetrazolium (NBT) reduction by white blood cells in normal, healthy, male subjects under non-fasting (before Ramadan) and fasting (after Ramadan) conditions. METHODS: In this study, 13 Muslim men, aged 28-54 years, whose health was confirmed by health application forms, gave blood samples one week before the beginning of the holy month of Ramadan and during the last week of Ramadan. Blood samples were tested for neutrophil phagocytosis, serum opsonisation power, and NBT reduction. RESULTS: Despite a decline in the neutrophil phagocytic index and serum opsonisation index, the percentage of neutrophils participating in phagocytosis increased with fasting. In addition, there was an increase in the percentage of neutrophils demonstrating NBT reduction. Although there was a decrease in opsonisation of the serum, the increased percentage of opsonisation compensated for this defect. CONCLUSION: This study demonstrates the beneficial effect of fasting during Ramadan on neutrophil phagocytic function.

Determination of the time required for appropriate chemical de-epithelialization of an ileal segment for cystoplasty: an animal model.

UNLABELLED: Another group of authors from Iran attempted experimentally to determine the required time for the appropriate enzymatic treatment of the ideal segment to complete de-epithelialization, thus reducing its absorptive function. They found that 25 min of enzymatic treatment of the ideal segment was adequate for this, and that it was recommended from their experimental study for cystoplasty. OBJECTIVES: To determine the time required for the appropriate enzymatic treatment of an ileal segment to de-epithelialize its mucosa and to reduce its absorptive function for cystoplasty in dogs. MATERIALS AND METHODS: Twenty-one adult female Persian mixed-breed dogs were divided into seven equal groups: group 1 (negative control group) had no ileocystoplasty; group 2 (positive control group) had a routine ileocystoplasty with no enzymatic treatment of the ileal segment; and groups 3-7 had an ileocystoplasty with 5, 10, 15, 20 or 25 min, respectively, of enzymatic treatment of the ileal segment with collagenase and trypsin. The seven groups were then compared for haematological, biochemical and histological changes, and glucose reabsorption assessed using a glucose-absorption test. RESULTS: No dogs showed any signs of metabolic disturbances, biochemical and haematological changes. There were significant differences in blood glucose level (BGL) for the groups at different times after the glucose-absorption test, but a pair-wise comparison showed significant differences in BGL between group 1 and the other groups, except group 7. The degree of histopathological change was associated with the duration of enzymatic treatment, in that changes were more prominent in group 7. There was no shrinkage or collagen deposition. CONCLUSIONS: In these conditions, 25 min of enzymatic treatment of the ileal segment is sufficient to remove the absorptive function of the augmented bladder, and is recommended for cystoplasty in dogs.