Puerarin Attenuates Obesity-Induced Inflammation and Dyslipidemia by Regulating Macrophages and TNF-Alpha in Obese Mice

Obesity causes low-grade inflammation that results in dyslipidemia and insulin resistance. We evaluated the effect of puerarin on obesity and metabolic complications both in silico and in vivo and investigated the underlying immunological mechanisms. Twenty C57BL/6 mice were divided into four groups: normal chow, control (HFD), HFD + puerarin (PUE) 200 mg/kg, and HFD + atorvastatin (ATO) 10 mg/kg groups. We examined bodyweight, oral glucose tolerance test, serum insulin, oral fat tolerance test, serum lipids, and adipocyte size. We also analyzed the percentage of total, M1, and M2 adipose tissue macrophages (ATMs) and the expression of F4/80, tumor necrosis factor-α (TNF-α), C-C motif chemokine ligand 2 (CCL2), CCL4, CCL5, and C-X-C motif chemokine receptor 4. In silico, we identified the treatment-targeted genes of puerarin and simulated molecular docking with puerarin and TNF, M1, and M2 macrophages based on functionally enriched pathways. Puerarin did not significantly change bodyweight but significantly improved fat pad weight, adipocyte size, fat area in the liver, free fatty acids, triglycerides, total cholesterol, and HDL-cholesterol in vivo. In addition, puerarin significantly decreased the ATM population and TNF-α expression. Therefore, puerarin is a potential anti-obesity treatment based on its anti-inflammatory effects in adipose tissue.

Improvement of Bovine Pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers.

Bovine pestiviruses are the causative agents of Bovine viral diarrhea, a disease that generates severe economic losses in cattle. The aim of this study was to improve its diagnosis by means of developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set a protocol for collecting, shipping and conserving bovine pestiviral RNA in filter papers. The developed RT-qPCR showed high sensitivity in detecting these viruses in different matrixes: viral stocks, semen and serum samples. Regarding the possibility of using the technique to test serum pools, it was possible to identify a positive serum sample within a pool containing 30 sera. In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovine samples was analyzed. The sampling method in two different filter papers using bovine blood drops was a useful alternative for diagnosis purposes and allowed the preservation of pestiviral RNA up to 12 months under refrigeration.

The Combined Effects of 6 Weeks of Jump Rope Interval Exercise and Dark Chocolate Consumption on Antioxidant Markers in Obese Adolescent Boys

Research has shown that both dark chocolate and exercise training may have favorable effects on antioxidant function in obese cohorts. However, their combined effect has not been established. We assessed the influences of six weeks of dark chocolate consumption combined with jump rope exercise on antioxidant markers in adolescent boys with obesity. Fifty adolescent boys with obesity (age = 15 ± 1 years) were randomly assigned into one of four groups; jump rope exercise + white chocolate consumption (JW; n = 13), jump rope exercise + dark chocolate consumption (JD; n = 13), dark chocolate consumption (DC; n = 12), or control (C; n = 12). Two participants dropped out of the study. Participants in JW and JD groups performed jump rope exercise three times per week for six weeks. Participants in the DC and JD groups consumed 30 g of dark chocolate containing 83% of cocoa during the same period. Serum concentrations of superoxide dismutase (SOD), total antioxidant capacity (TAC), glutathione peroxidase (GPx), and thiobarbituric acid reactive substances (TBARS) were evaluated prior to and after the interventions. All 3 intervention groups noted significant (p < 0.01) increases in serum concentrations of TAC, SOD, and GPx from baseline to post-test. In contrast, all intervention groups showed significantly reduced serum concentrations of TBARS from pre- to post-test (p ≤ 0.01). Bonferroni post hoc analysis revealed that post-test serum concentrations of TAC in the JD group were significantly greater than C (p < 0.001), DC (p = 0.010), and JW (p < 0.001) groups. In addition, post-test serum concentrations of SOD in the JD group were significantly greater than C group (p = 0.001). Post-test serum concentrations of GPx in the JD group were significantly greater than C (p < 0.001), DC (p = 0.021), and JW (p = 0.032) groups. The post-test serum concentrations of TBARS in the JD group was significantly lower than C (p < 0.001). No other significant between-group differences were observed. The current study provides evidence that dark chocolate consumption in combination with jump rope exercise is more efficient in improving antioxidant capacity than dark chocolate consumption or jump rope exercise alone among obese adolescent boys.

Sight-threatening progressive corneo-scleral involvement in porphyria cutanea tarda

Porphyria cutanea tarda is the most common type of porphyria. It is associated with a deficiency of uroporphyrinogen decarboxylase enzyme responsible for heme synthesis. Clinical manifestations are predominantly dermatological and very rarely present with ocular involvement. Although scleral thinning in the interpalpebral area is a well-documented entity, sight-threatening corneal involvement is rarely described. We, herein report a case of a 58-year-old man who presented with ocular surface dryness, photophobia and mild redness. Slit-lamp biomicroscopy revealed corneo-scleral thinning in both eyes. The diagnosis was confirmed with a urine porphyrin test, serum iron and serum ferritin levels. We started him on conservative management after which he was lost to follow-up. He presented again after 6 years with total corneal opacification and progressive loss of vision in the right eye.

Gynaecomastia in a male neutered cat with an adrenal tumour and associated hyperprogesteronism, hypercortisolism and hyperaldosteronism

Case summary A 7-year-old male neutered domestic longhair cat was presented with chronic progressive gynaecomastia, polydipsia, polyphagia, weight loss and poor fur regrowth. Sexualised behavioural changes were not reported and virilisation was not present on physical examination. Pertinent haematology, biochemistry and urinalysis findings at the time of referral included mild hypokalaemia. Left adrenomegaly and mild prostatomegaly were identified on a CT scan. Evaluation of adrenal hormones with a low-dose dexamethasone suppression test, serum progesterone, testosterone, oestradiol, plasma aldosterone, renin, plasma metanephrine and normetanephrine measurement supported a diagnosis of hyperprogesteronism, hyperaldosteronism and hypercortisolism. Adrenalectomy was performed and histopathology was consistent with an adrenocortical tumour. Clinical signs and hormone elevations resolved postoperatively. Relevance and novel information To our knowledge, this is the second report of gynaecomastia secondary to an adrenal tumour in a male neutered cat and the first associated with hyperprogesteronism.

Predicted reproductive longevity and women’s facial attractiveness

Physical attractiveness has been shown to reflect women’s current fecundity level, allowing a man to choose a potentially more fertile partner in mate choice context. However, women vary not only in terms of fecundity level at reproductive age but also in reproductive longevity, both influencing a couple’s long-term reproductive success. Thus, men should choose their potential partner not only based on cues of current fecundity but also on cues of reproductive longevity, and both may be reflected in women’s appearance. In this study, we investigated if a woman’s facial attractiveness at reproductive age reflects anti-Müllerian hormone (AMH) level, a hormone predictor of age at menopause, similarly as it reflects current fecundity level, estimated with estradiol level (E2). Face photographs of 183 healthy women (Mage = 28.49, SDage = 2.38), recruited between 2nd - 4th day of the menstrual cycle, were assessed by men in terms of attractiveness. Women’s health status was evaluated based on C-reactive protein level and biochemical blood test. Serum AMH and E2 were measured. The results showed that facial attractiveness was negatively correlated with AMH level, a hormone indicator of expected age at menopause, and positively with E2, indicator of current fecundity level, also when controlled for potential covariates (testosterone, BMI, age). This might result from biological trade-off between high fecundity and the length of reproductive lifespan in women and greater adaptive importance of high fecundity at reproductive age compared to the length of reproductive lifespan.

Discrepancy Between In-clinic and Haemagglutination-Inhibition Tests in Detecting Maternally-Derived Antibodies Against Canine Parvovirus in Puppies

Canine parvovirus (CPV) is one of the most common causes of mortality in puppies worldwide. Protection against CPV infection is based on vaccination, but maternally-derived antibodies (MDA) can interfere with vaccination. The aim of this study was to evaluate the applicability of an in-clinic ELISA test to assess the CPV MDA in unvaccinated puppies and CPV antibodies in bitches, comparing the results with the gold standard haemagglutination inhibition (HI) test. Serum samples of 136 unvaccinated puppies were tested, along with sera of 16 vaccinated bitches. Five unvaccinated puppies were retested after vaccination. Both assays showed that the 16 vaccinated bitches had protective antibody levels against CPV. Conversely, significant discrepancies were observed for the MDA titers in unvaccinated puppies. Protective MDA titers were observed in 91.9% puppies using HI and in 40.4% by the in-clinic ELISA test, and only the latter one showed a decrease of MDA titers and percentages of protected puppies after the first weeks of age. Vaccination of five puppies with high HI and low in-clinic ELISA MDA titers resulted in seroconversion. Our results confirm the reliability of the in-clinic ELISA test in determining protective antibodies against CPV in adult dogs. Our findings also suggest that the in-clinic ELISA test kit may also be a useful tool to detect and quantify CPV MDA, thus allowing prediction of the best time to vaccinate puppies and reduction of the rate of vaccination failures due to interference by maternally-derived antibodies.

Comparison of Syphilis Rapid Diagnostic Test to Rapid Plasma Reagin, Treponema pallidum Haemagglutination Assay and Fluorescent Treponemal Antibody-Absorption for Syphilis and Yaws Diagnostics

BACKGROUND: Syphilis and yaws are the treponemal infections which have become serious public health problems, and both are serologically indistinguishable. Developed serological tests for syphilis may also be used to diagnose yaws. In remote area, test modality with minimal requirements were needed. This study investigated the diagnostic value of syphilis rapid diagnostic test (RDT) in diagnosing syphilis and yaws.METHODS: For syphilis diagnostic test, serum samples were obtained from patients of outpatient clinic in Dr. Cipto Mangunkusumo National Central General Hospital who were sent for rapid plasma reagin (RPR) and Treponema pallidum haemagglutination assay (TPHA) to clinical laboratory of the hospital. The serum samples were collected and stored at -80°C until the day of testing for syphilis RDT and fluorescent treponemal antibody-absorption (FTA-Abs). For yaws diagnostic test, serum samples were obtained as a part of surveillance study of yaws among children 1-15 years old in West Halmahera. Venous blood samples were used for syphilis RDT and the sera were separated and were sent to Dr. Cipto Mangunkusumo National Central General Hospital for RPR, TPHA, and FTA-Abs tests.RESULTS: For syphilis diagnostic test, among 156 samples, 39 samples were positive with syphilis RDT. The sensitivity of syphilis RDT was similar to RPR and TPHA (100.0%), the specificity was same as TPHA (77.5%), but lower than RPR (84.8%) when compared to FTA-Abs IgM. The sensitivity of syphilis RDT was 62.5% and the specificity was 96.0% when compare to FTA-Abs IgG. For yaws diagnostic test, among 176 samples, 13 samples were positive with syphilis RDT. By using FTA-Abs IgM as gold standard for diagnosing yaws, the RDT have similar sensitivity (50.0%) with RPR and TPHA and syphilis RDT have similar specificity to TPHA (93.1%). If compared to FTA-Abs IgG, the sensitivity of syphilis RDT was 40.0% and the specificity was 98.0%.CONCLUSION: Syphilis RDT gives similar results with TPHA in syphilis and yaws cases. It may be used as a first line screening test latent or untreated syphilis and yaws because of good sensitivity. For yaws diagnosis Syphilis RDT, RPR, and TPHA have low sensitivity, however all those tests have an excellent agreement.KEYWORDS: FTA-Abs, rapid diagnostic test, syphilis, yaws

SARS-CoV-2 in first trimester pregnancy – does it affect the fetus?

BackgroundSeveral viral infections are known to be harmful to the fetus in the first trimester of pregnancy and can cause increased nuchal translucency thickness and pregnancy loss. Currently, no evidence exists regarding possible effects of SARS-CoV-2 in first trimester pregnancies.MethodsCohort 1 included pregnant women with a double test taken between Feb. 17 and Apr. 23, 2020, during the SARS-CoV-2 epidemic peak in Denmark. The double test was taken as part of the first trimester risk assessment. Cohort 2 included women with a first trimester pregnancy loss before double test. Serum from the double test or from a blood sample, in case of pregnancy loss, was analyzed for SARS-CoV-2 antibodies. The results were correlated to the nuchal translucency thickness and the number of pregnancy losses.ResultsIn total, 1,019 pregnant women with double test and 36 women with pregnancy loss participated in the study. Thirty (2.9%) women had SARS-CoV-2 antibodies in the serum from the double test. All women with pregnancy loss prior to the double test were negative for SARS-CoV-2 antibodies. There were no significant differences in nuchal translucency thickness for women testing positive (n=14) versus negative (p=0.20) or grey zone (n=16) versus negative (p=0.28). In total, 54 women experienced a pregnancy loss of whom two had grey zone or positive SARS-CoV-2 antibodies.ConclusionMaternal SARS-CoV-2 infection did not seem harmful in first trimester pregnancies. Infection had no effect on the nuchal translucency thickness and women with SARS-CoV-2 antibodies were not overrepresented among women with pregnancy loss.

Seroprevalence of Pre-Existing Nabs Against AAV1, 2, 5, 6 and 8 in South African Hemophilia B Patient Population

Introduction Several studies have shown that the induction of antibodies by natural exposure to various AAV serotypes can compromise the subsequent use of AAV as a gene therapy vector, limiting patient eligibility for AAV-delivered therapeutics. The implications of pre-existing antibodies to AAV serotypes are very different: Levels of anti-AAV2 or anti-AAV8 neutralizing antibodies (NABs) as low as 5 have been related to a decrease or even total impairment of AAV liver transduction after systemic delivery in humans. However, successful gene transfer has been reported in patients with anti-AAV5 NABs titers up to 340 and in non-human primates with titers up to 1030. Extensive surveys on the prevalence of anti-AAV antibodies in humans have been published. Results from these studies indicate that prevalence varies dependent on serotype, and that a significant proportion of individuals develop humoral immunity against various AAV serotypes early in life, starting around 2 years of age. Furthermore, the prevalence of antibodies to different AAV serotypes has been reported to vary according to geographical location. Study Objective We performed a NABs seroprevalence study in South African hemophilia B patient population (n=44) using a panel of AAV serotypes suitable for liver targeted therapy, to determine the AAV serotypes likely to be of greatest clinical applicability for the South African hemophilia B population. Methods Forty-four hemophilia B patient serum samples were obtained from Hemophilia Comprehensive Care Center in Johannesburg (South Africa). All the patient serum samples were analyzed for the presence of NABs against AAV serotypes 1, 2, 5, 6 and 8 with the use of highly sensitive luciferase-based bioassays. The assays entail incubation of the test serum samples dilution series with an AAV1, 2, 5, 6 or 8-based reporter vector that carries the luciferase gene. This incubation allows neutralizing antibodies in the test serum to bind to the reporter vector particles. These mixtures are subsequently transferred onto Hek293T cells, where reporter vector particles can transduce cells and mediate expression of luciferase. Anti-AAV1, 2, 5, 6 or 8 NABs titers were determined by calculation of the percentage of neutralization for each sample dilution and fitting the neutralization curve with a four-parameter method. Anti-AAV1, 2, 5, 6 or 8 NABs titer (IC50) is the dilution at which antibodies inhibit Hek293T cell transduction with AAV1, 2, 5, 6 or 8-LUC by 50%. The lowest patient serum dilution used in every assay was 8 and samples were considered positive when calculated anti-AAV NAB titer was ≥8. All analytical runs included proper negative and positive controls. Results and Discussion The presence of NABs against the AAV serotypes 1, 2, 5, 6 and 8 was determined in the serum of the hemophilia B patients (Fig. 1). The highest prevalence of NABs was found to be against the AAV2 serotype, 95% (n=42/44) followed by the AAV6 serotype, 82% (n=36/44), and the AAV1 serotype 77% (n=34/44). The prevalence of NABs against AAV5 and AAV8 was lower with 66% (n=29/44) for AAV5 and 64% (n=28/44) for AAV8. The serum samples positive for anti-AAV2 NABs had a high occurrence of titers above 1030 (39%) in comparison to anti-AAV1 NABs (20%), anti-AAV5 NABs (5%) or anti-AAV8 NABs (7%).The occurrence of samples with low titers (ranging from titer of 8 to titer of 50) was the highest for anti-AAV8 NABs (32%) and for anti-AAV5 NABs (27%), followed by anti-AAV2 (18%) and anti-AAV1 (5%) (Fig.1). Currently, an anti-AAV NABs titer of 5 is used as an exclusion criteria in most of the systemic AAV-based gene therapies. When applying a similar cut-off of 8, 23% of the analyzed patients could be treated with AAV1, 5% with AAV2, 34% with AAV5, 18% with AAV6 and 36% with AAV8-based therapeutics (Fig.2). However, we have previously reported that AAV5-neutralizing antibodies do not impair the efficacy of in vivo transduction of AAV5-based vector up to a measured titer of 340 in humans and 1030 in non-human primates. Therefore, applying the cut-off of 340 or 1030, either 84% or 95% of the South African Hemophilia B patients could benefit from treatment with AAV5-based gene therapy (Fig.2). Disclosures Majowicz: uniQure N.V.: Employment. van Waes:uniQure N.V.: Employment. Timmer:uniQure N.V.: Employment. van Deventer:uniQure Biopharma B.V.: Employment. Mahlangu:Takeda: Consultancy, Honoraria, Speakers Bureau; LFB: Consultancy; NovoNordisk: Consultancy, Research Funding, Speakers Bureau; Roche: Consultancy, Research Funding, Speakers Bureau; Baxalta: Consultancy, Research Funding, Speakers Bureau; Freeline Therapeutics: Research Funding; Pfizer: Consultancy, Research Funding, Speakers Bureau; Spark: Consultancy, Speakers Bureau; Chugai: Consultancy; Biomarin: Research Funding; CSL Behring: Consultancy, Research Funding, Speakers Bureau; Novartis: Research Funding; Sanofi Genzyme: Research Funding, Speakers Bureau; Shire: Consultancy, Research Funding, Speakers Bureau; Sobi: Research Funding, Speakers Bureau; uniQure: Research Funding; World Federation of Haemophilia: Speakers Bureau. Ferreira:uniQure N.V.: Employment.