Evaluation of Sex Differences in Murine Diabetic Ketoacidosis and Neutropenic Models of Invasive Mucormycosis

There is increased concern that the quality, generalizability and reproducibility of biomedical research can be influenced by the sex of animals used. We studied the differences between male and female mice in response to invasive pulmonary mucormycosis including susceptibility to infection, host immune reaction and responses to antifungal therapy. We used diabetic ketoacidotic (DKA) or neutropenic mice infected with either Rhizopus delemar or Mucor circinelloides. The only difference detected was that when DKA mice were infected with M. circinelloides, female mice were more resistant to infection than male mice (median survival time of 5 vs. 2 days for female and male mice, respectively). However, a 100% lethality was detected among infected animals of both sexes. Treatment with either liposomal amphotericin B (L-AMB) or posaconazole (POSA) protected mice from infection and eliminated the difference seen between infected but untreated female and male mice. Treatment with L-AMB consistently outperformed POSA in prolonging survival and reducing tissue fungal burden of DKA and neutropenic mice infected with R. delemar or M. circinelloides, in both mouse sexes. While little difference was detected in cytokine levels among both sexes, mucormycosis infection in the DKA mouse model induced more inflammatory cytokines/chemokines involved in neutrophil (CXCL1) and macrophage (CXCL2) recruitment vs. uninfected mice. As expected, this inflammatory response was reduced in the neutropenic mouse model. Our studies show that there are few differences between female and male DKA or neutropenic mice infected with mucormycosis with no effect on the outcome of treatment or host immune response.

Evaluation of Sex Differences in Murine Diabetic Ketoacidosis and Neutropenic Models of Invasive Mucormycosis

There is increased concern that the quality, generalizability, and reproducibility of biomedical research can be influenced by the sex of animals used. We studied the differences between male and female mice in response to invasive pulmonary mucormycosis including susceptibility to infection, host immune reaction, and responses to antifungal therapy. We used diabetic ketoacidotic (DKA) or neutropenic mice infected with either Rhizopus delemar or Mucor circinelloides. The only difference detected was that when DKA mice were infected with M. circinelloides, female mice were more resistant to infection than male mice (median survival time of 5 vs. 2 days for female and male mice, respectively). However, a 100% lethality was detected among infected animals of both sexes. Treatment with either liposomal amphotericin B (L-AMB) or posaconazole (POSA) protected mice from infection and eliminated the difference seen between infected but untreated female and male mice. Treatment with L-AMB consistently outperformed POSA in prolonging survival and reducing tissue fungal burden of DKA and neutropenic mice infected with R. delemar or M. circinelloides, in both mouse sexes. Although little difference was detected in cytokine levels among both sexes, mucormycosis infection in the DKA mouse model induced more inflammatory cytokines/chemokines involved in neutrophil (CXCL1) and macrophage (CXCL2) recruitment vs. uninfected mice. As expected, this inflammatory response was reduced in the neutropenic mouse model. Our studies show that there are few differences between female and male DKA or neutropenic mice infected with mucormycosis with no effect on the outcome of treatment or host immune response.

Effect of Lanthanum Strontium Manganese Oxide (LaSMnO3) Nanoparticle on mouse Testosterone and Fertility

Intraperitoneal administration of Lanthanum strontium manganese oxide (LaSMnO3) a new class of magnetic nanoparticle on mouse testosterone and fertility was investigated. For this, experimental mice divided into 4 groups (n=5); group I, II, III and IV were treated with vehicle (control), 5, 10 and 20 µg/kg/day of LaSMnO3 for 21 days respectively. Five animals from each group were sacrificed at interval of 0, 7, 14 and 21 days, however, after twenty-one days of the treatment, animals in all groups were allowed to cohabited with untreated female mice for fertility study. Toxic effects of LaSMnO3 on the testosterone and sperm parameters were analyzed. Effect on ROS and anti-oxidative biomarkers were also measured. Significant decrease (p<0.05) of epididymal spermatozoa motility and numbers was measured revealing the cytotoxicity effects of this nanomaterial. Light microscopic study revealed changes in the cauda epididymal sperm morphology. Failure of the fertility in LaSMnO3-treated mice as evidenced by the significant reduction in the average number of implantation in females mated with the treated males. Depletion of testicular testosterone hormone level by high dose of LaSMnO3 (20µg/kg/day) shows a reduced testicular androgen synthesis. This study therefore, shows the potential adverse effect of LaSMnO3 on male fertility. Keywords: Lanthanum strontium manganese oxide nanoparticle, animal models, toxicity, fertility

Investigating the Potential of Nigella Sativa and Thymoquinone in Salvaging the Embryo from Effects of Toxic Paternal Exposure to Cyclophosphamide

Background: Exposure to cyclophosphamide (CPA) for cancer treatment results in over-production of reactive oxygen species and oxidative stress thus affecting the DNA in male germ cell inducing sperm defects. Our goal is to assess the potential effects of Nigella sativa extract (NSE) and thymoquinone (TQ) on sperm and embryo quality following fertlization of sperm produced from germ cells which have been exposed to the damaging alkylating effects of CPA. Methods: Thirty six male ICR mice were divided into six groups; (I) Vehicle-treated control (normal saline), (II) CPA-only, (III) TQ-only, (IV) NSE-only, (V) CPA followed by TQ and (VI) CPA followed by NSE. Treatment with 200mg/kg CPA and 10mg/kg of both NSE and TQ were given by intraperitoneal injection. Animals were sacrificed at 33 days by cervical dislocation and sperm from caudal epidydymis were taken for analysis and in vitro fertilization (IVF) with eggs from untreated female. Fertilization rates and embryo development were monitored for 5 days. The result were analysed by using SPSS 16.Results: TQ and NSE supplementation to CPA-exposed male mice have no significant effect (p>0.05) on the total number of sperm if compared to CPA-only exposed mice. NSE and TQ supplementation have been shown to have significant effect (p<0.05) on the percentage of motile sperm as well as the number of abnormal sperm. Four types of abnormalities of the sperm were found which includes folded sperm, amorphous, banana-like and the head lacking of the usual hook. Finally, the embryo quality shows a significant improvement by the supplementation of TQ and NSE to CPA-exposed male mice (p<0.05). Conclusion: Overall, both NSE and TQ have indicated chemopreventive potential against the cytotoxicity of cyclophosphamide on the reproductive capacity and fertility.

MRI activity in MS and completed pregnancy

ObjectiveTo evaluate postpartum MRI activity in patients with MS and a completed pregnancy and to compare these results to an age-matched untreated nonpregnant MS cohort.MethodsPatient with MS from a tertiary care MS center between 2006 and 2015, with prepartum and postpartum neurologic follow-ups and MRI scans were analyzed. Clinical activity and inflammatory brain MRI activity (new T2-hyperintense or gadolinium-enhancing [Gd+] lesions) were assessed peripartum. The results were compared with untreated reproductive-age patients with MS from the placebo arm of the clinical trials.ResultsA total of 123 pregnancies in 123 women (median Expanded Disability Status Scale 1.0) were analyzed. Approximately 7.2% relapsed during pregnancy and 48.7% relapsed postpartum. Of pregnancies with prepartum and postpartum gadolinium (Gd)-enhanced MRI (n = 112), 8% had Gd+ lesions prepartum and 33% had new Gd+ lesions postpartum. Overall, 54.4% had either new T2 or Gd+ lesions postpartum. Seventy-nine percent of subjects with postpartum relapse had new MRI activity compared with 37.1% without relapse (p < 0.001). Twenty-five percent had both clinical and radiographic activity and only 24.9% maintained no evidence of disease activity status postpartum. There was no association between postpartum MRI activity and disease-modifying treatments (DMTs) (p > 0.5). MRI and clinical outcomes were also assessed for 126 nonpregnant untreated female patients with MS. Comparing pregnancy and no pregnancy groups, there was no difference in MRI activity at follow-up.ConclusionsThere was a high level of inflammatory radiographic disease activity which was related to relapses in postpartum patients with MS. Further studies are needed to determine whether hormonal fluctuations vs extended time off DMTs may be the underlying cause of our observations.

Transgenerational inheritance of abnormal spermatogenesis with Igf2/H19 epigenetic alteration in CD1 mouse induced by in utero arsenic exposure

Developmental exposure to environmental toxicants can induce transgenerational reproductive disease phenotypes through epigenetic mechanisms. However, little is known about the transgenerational effects of arsenic exposure. We hypothesize that prenatal arsenic exposure may result in impaired spermatogenesis in subsequent generations of male mice. To test our hypothesis, we treated pregnant CD-1 (F0) mice with drinking water containing sodium arsenite (85 ppm) from days 8 to 18 of gestation. Male offspring were bred with untreated female mice until the F3 generation was produced. Our results revealed that transient exposure of the F0 gestating female to arsenic can result in decreased sperm quality and histological abnormalities in testes of male offspring in the F1 and F3 generations. The overall methylation status of Igf2 DMR2 and H19 DMR was significantly lower in the arsenic-exposed group than that of the control group in both F1 and F3 generations. The relative mRNA expression levels of Igf2 and H19 in arsenic-exposed males were significantly higher than in the control males in both F1 and F3 generations. This study indicates that ancestral exposure to arsenic may result in transgenerational inheritance of an impaired spermatogenesis phenotyping involving both epigenetic alterations and the abnormal expression of Igf2 and H19.

Spontaneous clearance of Chlamydia trachomatis accounting for bacterial viability in vaginally or rectally infected women (FemCure)

ObjectivesSpontaneous clearance of Chlamydia trachomatis (CT) infections can occur between diagnosis and treatment. We followed CT patients to assess clearance using a conventional definition (no total CT-DNA, assessed by routine quantitative PCR methods) and a definition accounting for viability, assessed by viability PCR testing.MethodsThree outpatient STI clinics included CT-diagnosed women (The Netherlands, 2016–2017, FemCure study); participants had vaginal CT (vCT) and rectal CT (rCT) (group A: n=155), vCT and were rectally untested (group B: n=351), single vCT (group C: n=25) or single rCT (group D: n=29). Follow-up (median interval 9 days) vaginal and rectal samples underwent quantitative PCR testing (detecting total CT-DNA). When PCR positive, samples underwent V-PCR testing to detect ‘viable CT’ (CT-DNA from intact CT organisms; V-PCR positive). ‘Clearance’ was the proportion PCR-negative patients and ‘clearance of viable CT’ was the proportion of patients testing PCR negative or PCR positive but V-PCR negative. We used multivariable logistic regression analyses to assess diagnosis group (A–D), age, days since initial CT test (diagnosis) and study site (STI clinic) in relation to clearance and clearance of viable CT.ResultsClearance and clearance of viable CT at both anatomic sites were for (A) 0.6% and 3.9%; (B) 5.4% and 9.4%; (C) 32.0% and 52.0% and (D) 27.6% and 41.4%, respectively. In multivariate analyses, women with single infections (groups C and D) had higher likelihood of clearance than women concurrently infected with vCT and rCT (p<0.001).Of rectally untested women (group B), 76.9% had total CT-DNA and 46.7% had viable CT (V-PCR positive) at the rectal site.ConclusionsOf untreated female vCT patients who had CT also at the rectal site, or who were rectally untested, only a small proportion cleared CT (in fact many had viable CT) at their follow-up visit (median 9 days). Among single site infected women clearance was much higher.Trial registration numberNCT02694497.

Increased Serum Leptin Levels in New-Onset, Untreated Female Patients with Coronary Artery Disease and Positively Associated with Inflammatory Markers

Background/Aims: Previous studies have suggested that leptin was associated with atherosclerosis and involved in inflammation. Gender differences between leptin and inflammatory markers have been evaluated less in untreated patients with stable coronary artery disease (CAD). Methods: In this study, a total of 394 consecutive Chinese patients who received coronary artery angiography were enrolled, including 243 patients with CAD and 151 non-CAD controls. The baseline clinical characteristics were collected and serum leptin levels were determined using ELISA. Results: The relation of serum leptin levels to inflammatory markers was found only in female patients. Leptin and white blood cell count (WBCC) as well as its subsets were significantly higher in female patients than female controls. In female patients, leptin was positively associated with C-reactive protein (CRP; r = 0.28, p = 0.016), WBCC (r = 0.261, p = 0.02), neutrophil, r = 0.268, p = 0.018, and monocyte, r = 0.228, p = 0.044. Multivariable regression analysis revealed that leptin was significantly and independently associated with CRP (β = 0.317, p = 0.004), WBCC (β = 0.278, p = 0.020), neutrophil (β = 0.262, p = 0.032), and monocyte (β = 0.245, p = 0.032). Conclusions: The serum leptin levels were higher in female patients and independently associated with CRP, WBCC, and its subsets, suggesting a potential interaction between leptin and inflammation in female CAD patients.

Effect of feeding cottonseed cake on male fertility in rabbit

In order to study the effects of cottonseed cake on the fertility of the male rabbit, 60 animals (40 bucks and 20 does) were used. Males were 3 months old at the beginning of the essay, and the females were multiparous. Males were divided into 4 identical groups. Four diets containing 0, 6, 12 and 18% of cottonseed cake were formulated and randomly allotted to different groups of males. All the females received the same diet (6%). The treatment lasted for 90 days. At the end of that period, 5 males were sacrificed per lot and the rest were allowed to mate each with an untreated female before being sacrificed in turn. No significant difference (p>0.05) was observed among rations for the volume of testis and weight of reproductive organs. Histological study of the testes revealed no abnormality. In the presence of an untreated female, hundred percent of rabbits realized mounting and ejaculated whichever the cottonseed cake level. Nevertheless, the volume of the ejaculate was significantly (p>0.05) low in rabbit fed 6% cottonseed cake (0.86 vs 1.66, 12%). The spermatozoa concentration in the cauda epididymis was higher in rabbit given cottonseed cake than in control, although not significantly (p>0.05). The adverse effects of the toxicant contained in cottonseed cake occurred at 18% inclusion. The result showed that it is not possible to generalize the upper limit of inclusion of cottonseed cake in the ration formulation.DOI: http://dx.doi.org/10.3329/bjas.v44i1.23123 Bang. J. Anim. Sci. 2014. 44 (1): 16-20