DNA barcode reveals candidate species of Scinax and Ololygon (Anura: Hylidae) in Atlantic Forest

Molecular species delimitation methods are efficient tools to identify species, including the discovery of new taxa and cryptic organisms, thus being useful to biodiversity studies. In the present work, 16S mitochondrial sequences and cytochrome oxidase I (COI) were used to evaluate the richness of species in the genus Scinax and Ololygon from a biodiversity hotspot in Atlantic Forest. A total of 109 specimens formally belonging to eight species of Scinax and three species of Ololygon were collected in 13 localities along the state of Bahia (northeastern Brazil) and one site in Espírito Santo (southeastern Brazil). Of the Scinax species collected in this study, three were morphologically differentiated from other described species and identified as putative new species (Scinax sp.1, Scinax sp.2 and Scinax sp.3). The species delimitations were inferred using three different methods: ABGD, PTP and mPTP which allowed recognizing 11 Scinax species and five Ololygon species. Scinax sp. 1, Scinax sp. 2 and Scinax sp. 3, have been confirmed as new putative species and Ololygon argyreornata possibly contains cryptic species. We suggest additional studies, including morphological and bioacoustic data to validate these new putative species.     

ICOS Gene Polymorphisms (IVS1 + 173 T/C and c. 1624 C/T) in Primary Sjögren’s Syndrome Patients: Analysis of ICOS Expression

Background: Primary Sjögren’s syndrome (pSS) is a systemic autoimmune disease, which affects exocrine glands. T cell activation is a trigger mechanism in the immune response. Hyperreactivity of T cells and antibody production are features in pSS. ICOS can be critical in the pathogenesis of pSS. Methods: A total of 134 pSS patients and 134 control subjects (CS) were included. Genotyping was performed by PCR-RFLP. ICOS mRNA expression was quantified by real-time PCR, and CD4+ ICOS+ T cells were determined by flow cytometry. Results: The ICOS IVS1 + 173 T>C polymorphisms were not associated with susceptibility to pSS (p = 0.393, CI = 0.503–1.311). However, the c.1624 C>T polymorphism was associated with a reduction in the risk of development of pSS (p = 0.015, CI = 0.294–0.884). An increase in ICOS mRNA expression in patients was observed (3.7-fold). Furthermore, pSS patients showed an increase in membranal-ICOS expression (mICOS). High expression of mICOS (MFI) was associated with lymphocytic infiltration. Conclusions: The IVS1 + 173 polymorphism is not a genetic marker for the development of pSS, while c.1624 T allele was associated with a low risk. However, elevated mICOS expression in pSS patients with high lymphocytic infiltration was found. ICOS may have an important role in the immunopathogenesis of pSS and should be analyzed in T cell subsets in pSS patients as a possible disease marker.     

Prevention of violence against women and girls: A cost-effectiveness study across 6 low- and middle-income countries

BackgroundViolence against women and girls (VAWG) is a human rights violation with social, economic, and health consequences for survivors, perpetrators, and society. Robust evidence on economic, social, and health impact, plus the cost of delivery of VAWG prevention, is critical to making the case for investment, particularly in low- and middle-income countries (LMICs) where health sector resources are highly constrained. We report on the costs and health impact of VAWG prevention in 6 countries.Methods and findingsWe conducted a trial-based cost-effectiveness analysis of VAWG prevention interventions using primary data from 5 randomised controlled trials (RCTs) in sub-Saharan Africa and 1 in South Asia. We evaluated 2 school-based interventions aimed at adolescents (11 to 14 years old) and 2 workshop-based (small group or one to one) interventions, 1 community-based intervention, and 1 combined small group and community-based programme all aimed at adult men and women (18+ years old). All interventions were delivered between 2015 and 2018 and were compared to a do-nothing scenario, except for one of the school-based interventions (government-mandated programme) and for the combined intervention (access to financial services in small groups). We computed the health burden from VAWG with disability-adjusted life year (DALY). We estimated per capita DALYs averted using statistical models that reflect each trial’s design and any baseline imbalances. We report cost-effectiveness as cost per DALY averted and characterise uncertainty in the estimates with probabilistic sensitivity analysis (PSA) and cost-effectiveness acceptability curves (CEACs), which show the probability of cost-effectiveness at different thresholds. We report a subgroup analysis of the small group component of the combined intervention and no other subgroup analysis. We also report an impact inventory to illustrate interventions’ socioeconomic impact beyond health. We use a 3% discount rate for investment costs and a 1-year time horizon, assuming no effects post the intervention period. From a health sector perspective, the cost per DALY averted varies between US$222 (2018), for an established gender attitudes and harmful social norms change community-based intervention in Ghana, to US$17,548 (2018) for a livelihoods intervention in South Africa. Taking a societal perspective and including wider economic impact improves the cost-effectiveness of some interventions but reduces others. For example, interventions with positive economic impacts, often those with explicit economic goals, offset implementation costs and achieve more favourable cost-effectiveness ratios. Results are robust to sensitivity analyses. Our DALYs include a subset of the health consequences of VAWG exposure; we assume no mortality impact from any of the health consequences included in the DALYs calculations. In both cases, we may be underestimating overall health impact. We also do not report on participants’ health costs.ConclusionsWe demonstrate that investment in established community-based VAWG prevention interventions can improve population health in LMICs, even within highly constrained health budgets. However, several VAWG prevention interventions require further modification to achieve affordability and cost-effectiveness at scale. Broadening the range of social, health, and economic outcomes captured in future cost-effectiveness assessments remains critical to justifying the investment urgently required to prevent VAWG globally.

In a cost-effectiveness study, Dr. Giulia Ferrari and colleagues examine the costs and health impact of prevention of violence against women and girls in six low- and middle-income countries.     

Follow-up assessment of patients with Pure Neural Leprosy in a reference center in Rio de Janeiro—Brazil

IntroductionPure Neural Leprosy (PNL) is a rare clinical form of leprosy in which patients do not present with the classical skin lesions but have a high burden of the disability associated with the disease. Clinical characteristics and follow up of patients in PNL are still poorly described in the literature.ObjectiveThis paper aims to describe the clinical, electrophysiological and histopathological characteristics of PNL patients, as well as their evolution after multidrug therapy (MDT).MethodsFifty-two PNL patients were selected. Clinical, nerve conduction studies (NCS), histopathological and anti-PGL-1serology were evaluated. Patients were also assessed monthly during the MDT. At the end of the MDT, all of the patients had a new neurological examination and 44 were submitted to another NCS.ResultsParesthesia was the complaint most frequently reported by patients, and in the neurological examination the most common pattern observed was impairment in sensory and motor examination and a mononeuropathy multiplex. Painful nerve enlargement, a classical symptom of leprosy neuropathy, was observed in a minority of patients and in the motor NCS axonal injuries, alone or in combination with demyelinating features, were the most commonly observed. 88% of the patients did not present any leprosy reaction during MDT. There was no statistically significant difference between the neurological examinations, nor the NCS pattern, performed before and after the MDT.DiscussionThe classical hallmarks of leprosy neuropathy are not always present in PNL making the diagnosis even more challenging. Nerve biopsy is an important tool for PNL diagnosis as it may guide therapeutic decisions. This paper highlights unique characteristics of PNL in the spectrum of leprosy in an attempt to facilitate the diagnosis and management of these patients.     

Neosparton darwinii (Verbenaceae), a restricted endemic species. Is it also endangered?

Neosparton darwinii Benth. & Hook. is a ramose, almost leafless shrub, whose growth is restricted to an area on the southwestern coast of Buenos Aires province (Argentina). Its limited distribution could imply risks for its survival. In the present study, we evaluated population characteristics relevant to an assessment of its conservation status and made a preliminary characterization of its habitat. Historical data seem to indicate that the species has undergone a severe reduction in its geographic distribution. We estimated the actual population size to be around 44,000 plants, distributed in a clumped pattern. We found no significant differences between the environments immediately associated with the plants and other similar distant sites, in terms of position on the dune profile, percentage of bare soil and the richness and composition of accompanying plant species. A high proportion of the seeds produced each year is destroyed by parasite bruchid beetles. We have not seen seedlings of N.darwinii in the area. All the laboratory germination essays done to date have produced negative results. The species grows in an area intensively used by tourists. We can foresee quantitative and qualitative reductions in habitat availability. This information points out that N.darwinii is not just a rare species, but its survival is a matter for serious consideration. We suggest certain steps towards effective management to ensure its conservation.     

ATP dependence is not an intrinsic property of Na+/H+ exchanger NHE1: requirement for an ancillary factor

Na⁺/H⁺exchange is a passive process not requiring expenditure of metabolicenergy. Nevertheless, depletion of cellular ATP produces a markedinhibition of the antiport. No evidence has been found for directbinding of nucleotide to exchangers or alteration in their state ofphosphorylation, suggesting ancillary factors may be involved. Thispossibility was tested by comparing the activity of dog red blood cells(RBC) and their resealed ghosts. Immunoblotting experiments usingisoform-specific polyclonal and monoclonal antibodies indicated RBCmembranes expressNa⁺/H⁺exchanger isoform 1 (NHE1). In intact RBC, uptake ofNa⁺ was greatly stimulated whenthe cytosol was acidified. The stimulated uptake was largely eliminatedby amiloride and by submicromolar concentrations of the benzoylguanidinium compound HOE-694, consistent with mediation by NHE1.Although exchange activity could also be elicited by acidification inresealed ghosts containing ATP, the absolute rate of transport wasmarkedly diminished at comparable pH. Dissipation of the pH gradientwas ruled out as the cause of diminished transport rate in ghosts. Thiswas accomplished by a "pH clamping" procedure based on continuedexport of base equivalents by the endogenous anion exchanger. Theseobservations suggest a critical factor required to maintain optimalNa⁺/H⁺exchange activity is lost or inactivated during preparation of ghosts.Depletion of ATP, achieved by incubation with2-deoxy-D-glucose, inhibitedNa⁺/H⁺exchange in intact RBC, as reported for nucleated cells. In contrast, the rate of exchange was similar in control and ATP-depleted resealed ghosts. Interestingly, the residual rate ofNa⁺/H⁺exchange in ATP-depleted but otherwise intact cells was similar to thetransport rate of ghosts. Therefore, we tentatively conclude that fullactivation of NHE1 requires both ATP and an additional regulatoryfactor, which may mediate the action of the nucleotide. Ancillaryphosphoproteins or phospholipids or the kinases that mediate theirphosphorylation are likely candidates for the regulatory factor(s) thatis inactivated or missing in ghosts.

     

Interference of some tryptophan metabolites in the formation of melanin in vitro

Melanin (eumelanin) is commonly produced in mammals starting from tyrosine and/or 3,4-dioxyphenylalanine (DOPA) under the action of tyrosinase. 3-Hydroxyanthranilic acid and 3-hydroxykynurenine are intermediates occurring in the kynurenine pathway of tryptophan catabolism. In this paper, we show that these substances can interfere in melanin formation in vitro when tyrosine or DOPA is oxidized by molecular oxygen under catalysis by tyrosinase. In particular, when 3-hydroxyanthranilic acid is present, a brown and apparently water-soluble pigment is formed, whereas the typical eumelanin granules seem to become more and more rare as the concentration of 3-hydroxyanthranilic acid increases. Also in the presence of the latter, the rate of tyrosine and/or DOPA consumption decreases. A very complicated (13)C-NMR spectrum indicates the high complexity of the reaction. This involves both the true melanin precursor(s) and the tryptophan metabolite, even if with peculiar mechanism and kinetics. When 3-hydroxykynurenine is substituted for 3-hydroxyanthranilic acid the reaction leads to reddish pigments whereas xanthommatins (the typical oxidation products of 3-hydroxykynurenine) are absent. A possible relationship between some dischromic pathologies and tryptophan metabolic disorders is suggested.     

Stereoselectivity in the placental transfer and kinetic disposition of racemic bupivacaine administered to parturients with or without a vasoconstrictor

The aim of the present study was to investigate the stereoselectivity in the kinetic disposition and the transplacental distribution of bupivacaine in term parturients during labor. Maternal age ranged from 18-37 years and fetal gestational age from 37.6-41.5 weeks. Healthy parturients (n = 23) received epidural 0.5% racemic bupivacaine alone (group A) or combined with epinephrine (group B). Maternal venous blood was sampled at regular intervals until 8 h after drug administration and umbilical venous blood was obtained at delivery. Bupivacaine enantiomers were determined in plasma samples by HPLC using a Chiralcel(R) OD-R column and a UV detector. One- or two-compartment models were fitted to data and differences between the (+)-(R) and (-)-(S) enantiomers were compared with the paired Wilcoxon test (P< 0.05). The influence of epinephrine was evaluated using the unpaired Mann-Whitney test (P< 0.05). The disposition of bupivacaine in maternal plasma was stereoselective, with higher V(d/f) (140.60 vs. 132.81 L for group A and 197.86 vs. 169.46 L for group B) and C(l/f) (29.00 vs. 25.43 L/h for group A and 33.15 vs. 26.39 L/h for group B) and lower t(1/2)beta (3.24 vs. 3.30 h for group A and 4.36 vs. 4.45 h for group B) being observed for (+)-(R)-bupivacaine. The combined administration of epinephrine resulted in higher V(d/f) (197.86 vs. 140.60 L for (+)-(R) and 169.46 vs. 132.81 L for (-)-(S)) and t(1/2)beta values (4.36 vs. 3.24 h for (+)-(R) and 4.45 vs. 3.30 h for (-)-(S)). The transplacental distribution of bupivacaine was stereoselective only when bupivacaine was administered without epinephrine (group B), with a higher cord blood/maternal blood ratio being observed for (-)-(S)-bupivacaine (0.40 vs. 0.35). Chirality 16:65-71, 2004.