High resolution melting technique for molecular epidemiological studies of cystic echinococcosis: differentiating G1, G3, and G6 genotypes of Echinococcus granulosus sensu lato

Reliable and rapid genotyping of large number of Echinococcus granulosus sensu lato isolates is crucial for understanding the epidemiology and transmission of cystic echinococcosis. We have developed a method for distinguishing and discriminating common genotypes of E. granulosus s.l. (G1, G3, and G6) in Iran. This method is based on polymerase chain reaction coupled with high resolution melting curve (HRM), ramping from 70 to 86 °C with fluorescence data acquisition set at 0.1 °C increments and continuous fluorescence monitoring. Consistency of this technique was assessed by inter- and intra-assays. Assessment of intra- and inter-assay variability showed low and acceptable coefficient of variations ranging from 0.09 to 0.17 %. Two hundred and eighty E. granulosus s.l. isolates from sheep, cattle, and camel were used to evaluate the applicability and accuracy of the method. The isolates were categorized as G1 (93, 94, and 25 %), G3 (7, 4, and 4 %), and G6 (0, 2, and 71 %) for sheep, cattle, and camel, respectively. HRM results were completely compatible with those obtained from sequencing and rostellar hook measurement. This method proved to be a valuable screening tool for large-scale molecular epidemiological studies.     

The correlation between circulating inflammatory,oxidative stress,and neurotrophic factors level with the cognitive outcomes in multiple sclerosis patients

Neurological Sciences - Changes in the levels of circulating markers of inflammation, oxidative stress, and neurotrophic factors might be a good candidate for the prediction of cognitive impairment...     

Detection of Mycoplasma gallisepticum in experimentally infected broiler chickens using Culture, SPA, ELISA, and PCR methods

Avian mycoplasmosis is caused by several pathogenic mycoplasmas of which Mycoplasma gallisepticum and Mycoplasma synoviae are the most important. This experiment was designed to study the diagnosis of M. gallisepticum infection in broiler chicken receiving M. ga llisepticum. One hundred and twenty one-day-old chickens were raised in separate isolated rooms and equally allocated into two groups and renamed groups A and B were 121-day-old chickens. At 3 weeks old, group A was considered as the control group while group B was challenged with M. gallisepticum containing 1?×?10⁶ CFU/bird via an eye drop. In this study, culture, serum plate agglutination (SPA) test, the commercially available enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR) tests were compared for the detection of M. gallisepticum infection during the 1–7 weeks of age. Results revealed that M. gallisepticum was found in group B using culture and PCR during the 4–7-week-old period. The SPA results showed positive reactors in group B during the 5–7-week-old period. The positive reactors were detected by commercial ELISA test kit from 6 to 7 weeks of age. In this study, sensitivity of the PCR method was higher as compared to culture and serological tests. Its high sensitivity and fast turnaround time is making it the most frequently used test to monitor M. gallisepticum infection beside other tests.     

Non-aneurysmal non-perimesencephalic subarachnoid hemorrhage: effect of rehabilitation at short-term and in a prospective study of long-term follow-up

Background: A recently published prospective study identified an impaired outcome of patients with non-perimesencephalic (NPM) subarachnoid hemorrhage (SAH). Our objective was to analyze the long-term outcome of patients with subsequent rehabilitation after NPM SAH.

Methods: A comparison of patients with NPM SAH receiving subsequent in-patient rehabilitation was done at discharge (using the modified Rankin scale (mRS)), short-term outcome after 6 months (mRS), and prospectively using a questionnaire (short-form health survey with 36 questions (SF-36)), which was sent to 66 patients.

Results: Thirty-seven patients answered the SF-36, on average 6.3 years after ictus (range 1.5–14 years). After NPM SAH, the mRS is impaired. Patients with subsequent rehabilitation had a significant better improvement until short-term follow-up. Until long-term outcome, the psychological items were non-significantly reduced, whereas all physical items (physical functioning, role limitations because of physical health problems, bodily pain, and general health perceptions) were significantly decreased compared to the standard population. In patients with subsequent rehabilitation, all items were only non-significantly reduced. About 16% of the patients developed secondary neurological and/or psychiatric diseases.

Conclusions: The quality of life (QoL) is decreased after NPM SAH. In the long-term follow-up, a significant reduction in physical items was identified. Due to subsequent in-patient rehabilitation after NPM SAH, the impairment can be improved significantly until short-term follow-up. Whereas patients with NPM SAH had a significantly decreased QoL at long-term follow-up, for patients with rehabilitation, the QoL was only slightly (non-significantly) reduced. Therefore, patients should receive subsequent rehabilitation after NPM SAH to improve the functional short-term outcome (mRS) and long-term QoL. www.clinicaltrials.gov (Identifier No. NCT02334657).     

Serum hyaluronic acid and laminin as potential tumor markers for upper gastrointestinal cancers

Background

Early diagnosis of patients with upper gastrointestinal cancer is important because many cases are diagnosed in advanced stages and have poor prognosis. Several studies have reported increased serum levels of hyaluronic acid and laminin in various cancers and the correlation of the levels with poor prognosis. However, little data on the use of serum hyaluronic acid and laminin levels for early detection of esophageal and gastric cancers are available.

Methods

We assessed serum hyaluronic acid and laminin levels using enzyme-linked immunosorbent assay in 20 gastric cardia cancer, 23 gastric noncardia cancer and 20 esophageal squamous cell carcinoma incident cases and 25 controls in the Golestan Province, northern Iran, a high risk area for upper gastrointestinal cancers.

Results

Mean serum hyaluronic acid and laminin concentrations in cancer cases were higher than in controls in crude analyses. Significant correlations were observed between hyaluronic acid levels and gastric noncardia cancer (Beta-coefficient = 0.390; P = 0.01) and esophageal squamous cell carcinoma (Beta-coefficient = 0.332; P = 0.05) and between laminin levels and gastric cardia cancer (Beta-coefficient = 0.454; P = 0.003) in multivariate models. For esophageal squamous cell carcinoma, gastric cardia cancer, and gastric noncardia cancer, area under ROC curve (AUC) of hyaluronic acid was 0.708, 0.694, and 0.770, and of laminin was 0.706, 0.828, and 0.671.

Conclusions

Our study suggests that hyaluronic acid and laminin may be used to identify potentially high-risk groups of upper gastrointestinal cancers for further diagnostic work-ups, particularly in high incidence areas. Nevertheless, further studies with larger sample size and tumor staging information are warranted to clarify the clinical significance of hyaluronic acid and laminin in those cancers.     

Prevalence of horB gene among the Helicobacter pylori strains isolated from dyspeptic patients: first report from Iran

Helicobacter pylori (H. pylori) is globally accepted as an important cause of gastritis in human, and evidence strongly shows an etiological role for H. pylori in gastric cancer and peptic ulceration. In this study, we determined the relationship between digestive diseases and the horB gene of H. pylori infection. Fresh antral biopsy specimens were obtained from 140 dyspeptic patients (67 men and 73 women; mean age 41.5, aged 19?C63?years). They were examined for presence of the horB gene of H. pylori clinical isolates. Bacterial DNA content was extracted directly from the antral biopsy. Statistical analysis was performed with SPSS version 16.0. Prevalence of the horB gene in H. pylori isolated from patients with gastric cancer, gastric ulcer, gastritis and duodenal ulcer is (5/32) 15.6%, (4/25) 16%, (30/43) 70%, and (9/40) 22.5%, respectively. No significant relationship is observed between age, pathologic findings and gender factors with respect to the four digestive diseases (P?>?0.05). In our examination, a significant association was observed between a horB positive genotype of H. pylori and the occurrence of gastritis; in support of the protective theory. Studies with a higher sample size in different countries of the world should be conducted to obtain a thorough assessment as to whether horB has a role in the progress of gastritis (protective effect) or not. Further tests should be carried out to determine the exact role of horB in infection of H. pylori.     

Nodular lesions found in Irish poultry during veterinary inspection at poultrymeat plants

This investigation was carried out on three categories of old birds including cage layers, game birds (ducks and pheasants) and free-range birds. During the study 137,918 carcasses were examined grossly at slaughterhouses and 2040 condemned carcasses underwent detailed post-mortem examination. A total of 419 of the condemned carcasses had nodular lesions. On histological examination of the nodular lesions, one case of bacterial granuloma in a free-range layer, two cases of parasitic granulomata in ducks, 337 cases of different types of tumours and 79 cases of miscellaneous lesions with a nodular appearance were diagnosed. Overall, there was a total condemnation rate of 1.5%. The prevalence of various types of nodular lesions was discussed.     

Resistance in psychotherapy: what conclusions are supported by research.

Theoretical literature is contradictory in most areas, but virtually all theories agree on the existence of patient resistance and propose similar implications, meanings, and effects of its manifestation. However, theories differ widely in both the assumed causes of resistance and the methods of dealing with resistant patients. Common to various theoretical definitions is an assumption that resistance is both a dispositional trait and an in-therapy state of oppositional, angry, irritable, and suspicious behaviors. Reactance is a special class of resistance that is manifest in oppositional and uncooperative behavior. Resistance bodes poorly for treatment effectiveness. Nondirective and paradoxical strategies have been found to be quite successful in overcoming resistant and reactant states, while matching low-directive and self-directed treatments with resistant patients circumvents the effects of resistance traits.     

In vivo regeneration of bladder muscular wall using decellularized colon matrix: an experimental study

Background

Finding a proper scaffold for augmentation is a serious challenge in bladder tissue engineering. We hereby aimed to determine the histological aspects of a decellularized colon graft for bladder augmentation in healthy rats.

Methods

Rat colon tissues were decellularized using perfusion-based method. After partial cystectomy, bladders were grafted with a patch of decellularized colon. Bladder specimens were investigated in 12 rats at 1, 3, and 9 months postoperatively for further histological changes and immunohistochemistry analyses were also performed.

Results

One month after implantation, partial seeding of new cells was observed. After 3 months continuity of transitional epithelium of natural bladder on the decellularized grafted colon tissue was confirmed with histological and immunohistochemical examinations. All augmented bladders demonstrated a spherical shape without stone formation, necrosis or graft rejection. The presence of urothelium with similar morphology to the natural urothelium and visible blood vessels were found within 3 months of operation. All immunohistochemical markers (except markers of colonic stem cells) were expressed in biopsies obtained 3 months after surgery demonstrating a progressive vascular and smooth muscle cell infiltration into the graft after implantation.

Conclusion

This study suggests that decellularized colon may provide a viable material for bladder augmentation in rats to pave the road for future applications of this natural collagen scaffold.