rpoB gene mutations among Mycobacterium tuberculosis isolates from extrapulmonary sites

The aim of this study was to analyze mutations occurring in the rpoB gene of Mycobacterium tuberculosis (MTB) isolates from clinical samples of extrapulmonary tuberculosis (EPTB). Seventy formalin‐fixed, paraffin‐embedded samples and fresh tissue samples from confirmed EPTB cases were analyzed. Nested PCR based on the rpoB gene was performed on the extracted DNAs, combined with cloning and subsequent sequencing. Sixty‐seven (95.7%) samples were positive for nester PCR. Sequence analysis of the 81 bp region of the rpoB gene demonstrated mutations in 41 (61.2%) of 67 sequenced samples. Several point mutations including deletion mutations at codons 510, 512, 513 and 515, with 45% and 51% of the mutations in codons 512 and 513 respectively were seen, along with 26% replacement mutations at codons 509, 513, 514, 518, 520, 524 and 531. The most common alteration was Gln → His, at codon 513, presented in 30 (75.6%) isolates. This study demonstrated sequence alterations in codon 513 of the 81 bp region of the rpoB gene as the most common mutation occurred in 75.6% of molecularly confirmed rifampin‐resistant strains. In addition, simultaneous mutation at codons 512 and 513 was demonstrated in 34.3% of the isolates.     

The prevalence of celiac autoantibodies in hepatitis patients

Celiac disease has been associated with other autoimmune disorders such as autoimmune hepatitis, moreover it is known that T cell mediated immune response to dietary gluten and released cytokines are important for the entheropathy seen in celiac disease. We investigated celiac autoantibodies in patients with autoimmune hepatitis (AIH), and chronic hepatitis B (CHB).Sera from 84 patients with Autoimmune Hepatitis (AIH) type 1 and 88 patients with Chronic Hepatitis B (CHB) were tested for Immunoglobulin A and G antibodies to Gliadin, Immunoglobulin A antibodies to tissue transglutaminase using enzyme immunoassay, and Immunoglobulin A anti-endomysial antibodies by both indirect immunofluorescence, and enzyme immunoassay. The patients positive for anti-endomysial antibodies and/or anti tissue transglutaminase antibodies were considered for deuodenal biopsy. The study was approved by Research Center for Gastroenterology and Liver Disease Ethics Committee and all patients gave their written informed consent to participate.Immunoglobulin A anti-endomysial and Immunoglobulin A anti-gliadin antibodies were positive in two out of 84 patients with AIH. Moreover, Immunoglobulin A anti-gliadin antibodies were positive in another patient who was also positive for anti tissue transglutaminase antibodies. Tissue transglutaminase antibodies were positive in eight (9.1%) of 88 patients with CHB, two of which were also positive for anti-endomysial antibodies. One of the patients with CHB was only positive for anti-endomysial antibodies.Compared with the general population, the prevalence of celiac autoantibodies in CHB and AIH patients is relatively high, and it is noteworthy that most positive patients were asymptomatic for celiac disease. We suggest screening for celiac disease before and during treatment in patients with viral and autoimmune hepatitis.     

Cutaneous manifestations of diabetes mellitus: a case series

Diabetes mellitus (DM) is a common disorder with a broad spectrum of cutaneous manifestations. Our purpose was to evaluate the prevalence and main clinical presentation of skin disorders in patients with DM. For a period of 6 months, all of the patients with DM attending the outpatient dermatology and diabetes clinics of the Hamedan University of Medical Sciences, Iran, were clinically examined for cutaneous manifestations of DM. Patients also were evaluated for glycemic control and evidence of other diabetes-related complications. Diabetic skin manifestations were detected in 110 of 155 (71%) patients with DM. The most common skin lesions in both patients with type 1 and type 2 DM were infectious in origin (72%). No statistically significant differences in cutaneous manifestations were observed between the 2 types of DM. In the outpatient population with DM there is a high prevalence of skin lesions mainly represented by cutaneous infections.     

Modulation of morphine state-dependent learning by muscarinic cholinergic receptors of the ventral tegmental area

In the present study, the effects of bilateral intra-ventral tegmental area (intra-VTA) injections of an anticholinesterase, physostigmine and/or muscarinic acetylcholine receptor antagonist, atropine on memory retention and morphine state-dependent learning were examined in adult male Wistar rats. As a model of learning, a step-through passive avoidance task was used. Post-training subcutaneous administration of morphine (0.5, 2.5 and 5 mg/kg) dose-dependently impaired memory retrieval on the test day. Pre-test administration of morphine (2.5 and 5 mg/kg) induced state-dependent retrieval of the memory acquired under post-training morphine influence. Pre-test intra-VTA microinjection of physostigmine (0.5, 1 and 2 microg/rat) or atropine (1, 2 and 3 microg/rat) alone cannot affect memory retention. Interestingly, pre-test intra-VTA administration of physostigmine (1 and 2 microg/rat) reversed post-training morphine (5 mg/kg, s.c.)-induced retrieval impairment. Furthermore, pre-test intra-VTA microinjection of physostigmine (1 and 2 mug/rat) with an ineffective dose of morphine (0.5 mg/kg), synergistically improved memory performance impaired by post-training morphine. On the other hand, pre-test intra-VTA microinjection of atropine (2 and 3 microg/rat) 5 min before the administration of morphine (5 mg/kg, s.c.) dose-dependently inhibited morphine state-dependent memory. Pre-test atropine microinjection also reversed the influence of physostigmine on morphine response. It may be concluded that the muscarinic acetylcholine receptors of the VTA play an important role in morphine-induced recovery of memory, on the test day.     

GABA(A) receptors in the basolateral amygdala are involved in mediating morphine reward

In the present study, the effects of intra-basolateral amygdala (BLA) injection of GABA(A) receptor agonist and antagonist on morphine-induced conditioned place preference (CPP) in male Wistar rats have been investigated. Subcutaneous (s.c.) administration of different doses of morphine sulfate (1-9 mg/kg) produced a dose-dependent CPP. Using a 3-day schedule of conditioning, it was found that the GABA(A) receptor agonist, muscimol (0.125, 0.25 and 0.5 microg/rat) or the GABA(A) receptor antagonist, bicuculline (0.125, 0.25 and 0.5 microg/rat), did not produce a significant place preference or place aversion. Intra-BLA administration of muscimol (0.25 and 0.5 microg/rat) decreased the acquisition of CPP induced by morphine (6 mg/kg). On the other hand, intra-BLA injection of bicuculline (0.25 and 0.5 microg/rat) in combination with an ineffective dose of morphine (1 mg/kg) elicited a significant CPP. The response of different doses of muscimol was attenuated by bicuculline (0.125 and 0.25 microg/rat). Furthermore, intra-BLA administration of bicuculline but not muscimol before testing significantly decreased the expression of morphine (6 mg/kg)-induced place preference. The administration of the higher doses of bicuculline (0.25 and 0.5 microg/rat) during acquisition and the higher dose of muscimol (2 microg/rat) on the test day decreased the locomotor activity of the animals on the testing phase. It can be concluded that GABA(A) receptors in the amygdala are involved in morphine reward.     

Inhibition of morphine-induced amnesia in morphine-sensitized mice: involvement of dorsal hippocampal GABAergic receptors

In the present study, the effects of bilateral injections of the GABAergic receptor agents into the dorsal hippocampal CA1 regions (intra-CA1) on morphine-induced amnesia were examined in morphine sensitized-mice. Pre-training subcutaneous (s.c.) administration of morphine (5 mg/kg) suppressed the learning of a one-trial passive avoidance task. Amnesia induced by pre-training morphine was significantly reversed in mice which had previously received once daily injections of morphine (20 and 30 mg/kg, s.c.) for 3 days, which may be due to behavioral sensitization. Intra-CA1 injections of GABAA receptor agonist, muscimol (0.125, 0.25, 0.5 and 1 microg/mouse) or the GABAB receptor agonist, baclofen (1, 1.5 and 2 microg/mouse) prior to injection of morphine (20 mg/kg per dayx3 days) decreased the reversion of morphine-induced amnesia in morphine sensitized-mice. Daily intra-CA1 injections of muscimol or baclofen plus saline for 3 days did not alter memory formation. Furthermore, during development of sensitization, the combination of GABAA receptor antagonist, bicuculline (0.25, 0.5, 1 and 2 microg/mouse) with an ineffective dose of morphine (5 mg/kg, s.c.) reversed the amnesia induced by pre-training morphine. However, the same treatment with GABAB receptor antagonist, CGP35348 (2.5-40 microg/mouse) had no effect on the morphine response. On the other hand, daily intra-CA1 injections of bicuculline or CGP35348 alone for 3 days did not alter the amnesia induced by pre-training injection of morphine. The results suggest that morphine sensitization reverses the impairment of memory induced by morphine and that GABAergic receptors of the dorsal hippocampus may play an important role in this effect.     

Fe3O4@Sap/Cu(ii): an efficient magnetically recoverable green nanocatalyst for the preparation of acridine and quinazoline derivatives in aqueous media at room temperature

Saponin, as a green and available phytochemical, was immobilized on the surface of magnetite nanoparticles then doped with Cu ions (Fe₃O₄@Sap/Cu(ii)) and used as an efficient nanocatalyst for the synthesis of quinazoline and acridine derivatives, due to their high application and importance in various fields of science. Different spectroscopic and microscopic techniques were used for the catalyst characterization such as FT-IR, XRD, FE-SEM, EDX, TEM, TGA, VSM, BET, DLS, CV, and XPS analyses. All characterization data were correlated with each other so that the structure of the catalyst was accurately characterized. The reactions were performed in the presence of a low amount of Fe₃O₄@Sap/Cu(ii) (0.42 mol%) as a green catalyst in water over a short period of time. The results show well the effective role of saponin in solving the problem of mass transfer in aqueous medium, which is the challenge of many organic reactions in aqueous medium and in the presence of heterogeneous medium. High catalytic activity was found for the catalyst and high to excellent efficiency was obtained for all quinazoline (68–94% yield) and acridine (66–97% yield) derivatives in short reaction times (less than 1 hour) under mild reaction conditions in the absence of any hazardous or expensive materials. There is not any noticeable by-product found whether for acridine or quinazoline derivatives, which reflects the high selectivity. Two reasonable mechanisms were proposed for the reactions based on observations from control experiments as well as literature reports. The catalyst could be easily recovered magnetically for at least six consecutive runs with insignificant reactivity loss.

A highly efficient, robust, and green protocol has been developed for the synthesis of acridine and quinazoline derivatives in water under mild reaction conditions using a Fe₃O₄@Sap/Cu(ii) nanocomposite as an efficient heterogeneous catalyst.     

Prevalence of and Risk Factors for Genito-Pelvic Pain/Penetration Disorder: A Population-Based Study of Iranian Women

IntroductionTo date, few studies have investigated the prevalence of sexual pain in the context of the new diagnostic concept of genito-pelvic pain/penetration disorder (GPPPD).AimTo evaluate the prevalence of GPPPD and its associated factors.MethodsThis was a population-based, cross-sectional study of 590 healthy married women age 18–70 years conducted between May and October 2017 in Tehran, Iran.Main Outcome MeasuresResearch tools included demographic characteristics checklist, factors affecting GPPPD, sexual distress and self-reporting of pain during intercourse, 2 standard questionnaires on depression (Patient Health Questionnaire 9) and Binik’s guideline for the diagnosis of GPPPD.Results196 women (33%) reported pain or fear in answer to self-report questions. Administration of Binik’s guideline yielded a GPPPD prevalence of 16% (n = 94 women); however, this number decreased to 62 women (10.5%) when sexual distress was taken into account; thus, the final prevalence of GPPPD was considered to be 10.5%. However, if the threshold in Binik’s guideline was lowered to also include those reporting “somewhat” pain in addition to the group reporting “moderate” and “quite a bit or always,” then the prevalence of GPPPD increased to 25.8%. The results of backward logistic regression identified a strong aversion to looking at or touching the genitalia (odd ratio [OR] = 4.3), low sexual satisfaction (OR = 3.1), and severe depression (OR = 6.6) as independent risk factors for a diagnosis of GPPPD and secure financial status (OR = 0.3) and a high level of marital satisfaction (OR = 0.2) as protective factors against a diagnosis of GPPPD.Clinical ImplicationsReliable diagnosis of GPPPD is crucial. Application of validated tools may mitigate the overestimation of GPPPD prevalence. Simultaneously, clinicians’ judgment is essential in assessing a reasonable threshold and preventing underestimation that leads to the exclusion of women suffering from pain.Strengths & LimitationsThe present study is one of the few evaluating the prevalence of GPPPD according to the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-5) definition and Binik’s guideline. The study also aims to point out some new perspectives on merging the 2 concepts of vaginismus and dyspareunia. Study limitations include the evaluation of factors affecting GPPPD based on self-reporting and possible recall bias.ConclusionFurther research is needed to determine the appropriate threshold for a diagnosis of GPPPD. We suggest that a woman with mild to moderate pain or fear of vaginal penetration is under sexual distress and cannot be neglected. In addition, problems may arise following the DSM-5 merging of the 2 disorders of vaginismus and dyspareunia, owing to the significant prevalence and distress of lifelong vaginismus in some cultures.Alizadeh A, Farnam F, Raisi F, et al. Prevalence of and Risk Factors for Genito-Pelvic Pain/Penetration Disorder: A Population-Based Study of Iranian Women.J Sex Med 2019;16:1068–1077.     

Effects of <Emphasis Type="Italic">Ginkgo biloba</Emphasis> on cerebral blood flow assessed by quantitative MR perfusion imaging: a pilot study

Introduction  

Extract of Ginkgo biloba (EGb), a dietary supplement used for a number of conditions including dementia, has been suggested to increase cerebral blood flow (CBF). The purpose of this study was to determine if changes in CBF could be detected by dynamic susceptibility contrast-enhanced magnetic resonance imaging (DSC-MRI) in elderly human subjects taking EGb.