Micromanipulation at an infertility centre

Aim. Human in vitro fertilisation (IVF) and gamete intrafallopian transfer have been used in the management of various forms of infertility. In cases of severe male-factor infertility, fertilisation can be a factor. In this study micromanipulation was used to increase fertilisation in such cases.Methods. Two micromanipulation techniques, subzonal sperm injection (SUZI) and partial zona dissection (PZD), were used to assist fertilisation in patients with abnorTl)al semen parameters. Ten couples with severe oligo-, teratoand asthenozoospermia participated in the SUZI programme. Seventy-three oocytes were obtained from these 10 patients. PZD was used on day 1 oocytes in cases of male infertility as well as a rescue attempt on day 2 oocytes when fertilisation had failed after routine insemination.Results. The SUZI technique had a fertilisation rate of 37,7%. In this group, a biochemical pregnancy was achieved. Differences between the fertilisation rate of conventionallVF (33,3%) and PZD (56,3%) in cases of male infertility, were not statistically significant although a clinical difference could be detected. PZD was statistically effective in facilitating fertilisation (37,5% v. 8,3%) in couples where this procedure was introduced to reinseminate 24-hour-old unfertilised oocytes. Four patients received PZD reinseminated embryos. An average of 1,45 PZD embryos were replaced and 1 implantation pregnancy was confirmed.Conclusion. The micromanipulation results are encouraging arid seemed to increase the efficiency of IVF in humans. Furthermore, our data support the conclusion that micromanipulation procedures can bring about pregnancies

Bioavailability of Microplastics to Marine Zooplankton: Effect of Shape and Infochemicals

The underlying mechanisms that influence microplastic ingestion in marine zooplankton remain poorly understood. Here, we investigate how microplastics of a variety of shapes (bead, fiber, and fragment), in combination with the algal-derived infochemicals dimethyl sulfide (DMS) and dimethylsulfoniopropionate (DMSP), affect the ingestion rate of microplastics in three species of zooplankton, the copepods Calanus helgolandicus and Acartia tonsa and larvae of the European lobster Homarus gammarus. We show that shape affects microplastic bioavailability to different species of zooplankton, with each species ingesting significantly more of a certain shape: C. helgolandicus—fragments (P < 0.05); A. tonsa—fibers (P < 0.01); H. gammarus larvae—beads (P < 0.05). Thus, different feeding strategies between species may affect shape selectivity. Our results also showed significantly increased ingestion rates by C. helgolandicus on all microplastics that were infused with DMS (P < 0.01) and by H. gammarus larvae and A. tonsa on DMS-infused fibers and fragments (P < 0.05). By using a range of more environmentally relevant microplastics, our findings highlight how the feeding strategies of different zooplankton species may influence their susceptibility to microplastic ingestion. Furthermore, our novel study suggests that species reliant on chemosensory cues to locate their prey may be at an increased risk of ingesting aged microplastics in the marine environment

Distribution and Molecular Evolution of Bacillus anthracis Genotypes in Namibia

The recent development of genetic markers for Bacillus anthracis has made it possible to monitor the spread and distribution of this pathogen during and between anthrax outbreaks. In Namibia, anthrax outbreaks occur annually in the Etosha National Park (ENP) and on private game and livestock farms. We genotyped 384 B. anthracis isolates collected between 1983–2010 to identify the possible epidemiological correlations of anthrax outbreaks within and outside the ENP and to analyze genetic relationships between isolates from domestic and wild animals. The isolates came from 20 animal species and from the environment and were genotyped using a 31-marker multi-locus-VNTR-analysis (MLVA) and, in part, by twelve single nucleotide polymorphism (SNP) markers and four single nucleotide repeat (SNR) markers. A total of 37 genotypes (GT) were identified by MLVA, belonging to four SNP-groups. All GTs belonged to the A-branch in the cluster- and SNP-analyses. Thirteen GTs were found only outside the ENP, 18 only within the ENP and 6 both inside and outside. Genetic distances between isolates increased with increasing time between isolations. However, genetic distance between isolates at the beginning and end of the study period was relatively small, indicating that while the majority of GTs were only found sporadically, three genetically close GTs, accounting for more than four fifths of all the ENP isolates, appeared dominant throughout the study period. Genetic distances among isolates were significantly greater for isolates from different host species, but this effect was small, suggesting that while species-specific ecological factors may affect exposure processes, transmission cycles in different host species are still highly interrelated. The MLVA data were further used to establish a model of the probable evolution of GTs within the endemic region of the ENP. SNR-analysis was helpful in correlating an isolate with its source but did not elucidate epidemiological relationships

A Single-Arm, Proof-Of-Concept Trial of Lopimune (Lopinavir/Ritonavir) as a Treatment for HPV-Related Pre-Invasive Cervical Disease

BACKGROUND: Cervical cancer is the most common female malignancy in the developing nations and the third most common cancer in women globally. An effective, inexpensive and self-applied topical treatment would be an ideal solution for treatment of screen-detected, pre-invasive cervical disease in low resource settings. METHODS: Between 01/03/2013 and 01/08/2013, women attending Kenyatta National Hospital's Family Planning and Gynaecology Outpatients clinics were tested for HIV, HPV (Cervista®) and liquid based cervical cytology (LBC -ThinPrep®). HIV negative women diagnosed as high-risk HPV positive with high grade squamous intraepithelial lesions (HSIL) were examined by colposcopy and given a 2 week course of 1 capsule of Lopimune (CIPLA) twice daily, to be self-applied as a vaginal pessary. Colposcopy, HPV testing and LBC were repeated at 4 and 12 weeks post-start of treatment with a final punch biopsy at 3 months for histology. Primary outcome measures were acceptability of treatment with efficacy as a secondary consideration. RESULTS: A total of 23 women with HSIL were treated with Lopimune during which time no adverse reactions were reported. A maximum concentration of 10 ng/ml of lopinavir was detected in patient plasma 1 week after starting treatment. HPV was no longer detected in 12/23 (52.2%, 95%CI: 30.6-73.2%). Post-treatment cytology at 12 weeks on women with HSIL, showed 14/22 (63.6%, 95%CI: 40.6-82.8%) had no dysplasia and 4/22 (18.2%, 95%CI: 9.9-65.1%) were now low grade demonstrating a combined positive response in 81.8% of women of which 77.8% was confirmed by histology. These data are supported by colposcopic images, which show regression of cervical lesions. CONCLUSIONS: These results demonstrate the potential of Lopimune as a self-applied therapy for HPV infection and related cervical lesions. Since there were no serious adverse events or detectable post-treatment morbidity, this study indicates that further trials are clearly justified to define optimal regimes and the overall benefit of this therapy. TRIAL REGISTRATION: ISRCTN Registry 48776874

Persistent anthrax as a major driver of wildlife mortality in a tropical rainforest

Anthrax is a globally important animal disease and zoonosis. Despite this, our current knowledge of anthrax ecology is largely limited to arid ecosystems, where outbreaks are most commonly reported. Here we show that the dynamics of an anthrax-causing agent, Bacillus cereus biovar anthracis, in a tropical rainforest have severe consequences for local wildlife communities. Using data and samples collected over three decades, we show that rainforest anthrax is a persistent and widespread cause of death for a broad range of mammalian hosts. We predict that this pathogen will accelerate the decline and possibly result in the extirpation of local chimpanzee (Pan troglodytes verus) populations. We present the epidemiology of a cryptic pathogen and show that its presence has important implications for conservation

Activation of P2X7-mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

<p>Abstract</p> <p>Background</p> <p>The study tested the hypothesis that apoptosis can prevent and control growth of neoplastic cells. Previous studies in-vitro have shown that the pro-apoptotic P2X₇receptor regulates growth of epithelial cells. The specific objective of the present study was to understand to what degree the P2X₇system controls development and growth of skin cancer in vivo, and what cellular and molecular mechanisms are involved in the P2X₇action.</p> <p>Methods</p> <p>Skin neoplasias in mice (papillomas, followed by squamous spindle-cell carcinomas) were induced by local application of DMBA/TPA. Experiments in-vitro utilized cultured epidermal keratinocytes generated from wild-type or from P2X₇-null mice. Assays involved protein immunostaining and Western blots; mRNA real-time qPCR; and apoptosis (evaluated in situ by TUNEL and quantified in cultured keratinocytes as solubilized DNA or by ELISA). Changes in cytosolic calcium or in ethidium bromide influx (P2X₇pore formation) were determined by confocal laser microscopy.</p> <p>Results</p> <p>(a) Co-application on the skin of the P2X₇specific agonist BzATP inhibited formation of DMBA/TPA-induced skin papillomas and carcinomas. At the completion of study (week 28) the proportion of living animals with cancers in the DMBA/TPA group was 100% compared to 43% in the DMBA/TPA+BzATP group. (b) In the normal skin BzATP affected mainly P2X₇-receptor – expressing proliferating keratinocytes, where it augmented apoptosis without evoking inflammatory changes. (c) In BzATP-treated mice the degree of apoptosis was lesser in cancer than in normal or papilloma keratinocytes. (d) Levels of P2X₇receptor, protein and mRNA were 4–5 fold lower in cancer tissues than in normal mouse tissues. (e) In cultured mouse keratinocytes BzATP induced apoptosis, formation of pores in the plasma membrane, and facilitated prolonged calcium influx. (f) The BzATP-induced apoptosis, pore-formation and augmented calcium influx had similar dose-dependence for BzATP. (g) Pore formation and the augmented calcium influx were depended on the expression of the P2X₇receptor, while the BzATP-induced apoptosis depended on calcium influx. (h) The BzATP-induced apoptosis could be blocked by co-treatment with inhibitors of caspase-9 and caspase-3, but not of caspase-8.</p> <p>Conclusion</p> <p>(a) P2X₇-dependent apoptosis is an important mechanism that controls the development and progression of epidermal neoplasia in the mouse. (b) The P2X₇-dependent apoptosis is mediated by calcium influx via P2X₇pores, and involves the caspase-9 (mitochondrial) pathway. (c) The diminished pro-apoptotic effect of BzATP in mouse cancer keratinocytes is possibly the result of low expression of the P2X₇receptor. (d) Activation of P2X₇-dependent apoptosis, e.g. with BzATP could be a novel chemotherapeutic growth-preventive modality for papillomas and epithelial cancers in vivo.</p

Population genomics of marine zooplankton

Author Posting. © The Author(s), 2017. This is the author's version of the work. It is posted here for personal use, not for redistribution. The definitive version was published in Bucklin, Ann et al. "Population Genomics of Marine Zooplankton." Population Genomics: Marine Organisms. Ed. Om P. Rajora and Marjorie Oleksiak. Springer, 2018. doi:10.1007/13836_2017_9.The exceptionally large population size and cosmopolitan biogeographic distribution that distinguish many – but not all – marine zooplankton species generate similarly exceptional patterns of population genetic and genomic diversity and structure. The phylogenetic diversity of zooplankton has slowed the application of population genomic approaches, due to lack of genomic resources for closelyrelated species and diversity of genomic architecture, including highly-replicated genomes of many crustaceans. Use of numerous genomic markers, especially single nucleotide polymorphisms (SNPs), is transforming our ability to analyze population genetics and connectivity of marine zooplankton, and providing new understanding and different answers than earlier analyses, which typically used mitochondrial DNA and microsatellite markers. Population genomic approaches have confirmed that, despite high dispersal potential, many zooplankton species exhibit genetic structuring among geographic populations, especially at large ocean-basin scales, and have revealed patterns and pathways of population connectivity that do not always track ocean circulation. Genomic and transcriptomic resources are critically needed to allow further examination of micro-evolution and local adaptation, including identification of genes that show evidence of selection. These new tools will also enable further examination of the significance of small-scale genetic heterogeneity of marine zooplankton, to discriminate genetic “noise” in large and patchy populations from local adaptation to environmental conditions and change.Support was provided by the US National Science Foundation to AB and RJO (PLR-1044982) and to RJO (MCB-1613856); support to IS and MC was provided by Nord University (Norway)

Palinspastic restoration of an exhumed deep-water system: a workflow to improve paleogeographic reconstructions

The Permian Laingsburg depocenter, Karoo Basin, South Africa is the focus of sedimentological and stratigraphic research as an exhumed analogue for offshore hydrocarbon reservoirs in deep-water basins. Thin-skinned thrust tectonics during the Permo-Triassic Cape Orogeny resulted in post-depositional deformation of the Permian basin-fill. Regional-scale cross sections reveal two structural domains: a southern domain in the Laingsburg depocenter comprising 8-11 km wavelength north-verging fault-propagation folding, driven by buried low angle (<45o) reverse faults that coalesce at depth as part of a mega-detachment below the Lower Paleozoic Cape Supergroup; and a northern domain to the north of the Laingsburg depocenter of short wavelength, low amplitude, asymmetrical folding facilitated by a detachment within the Permian Ecca Group. Five detailed structural cross sections permit the palinspastic restoration, and a calculation of the amount of shortening, across a 2500 km2 area in the Laingsburg depocenter. Average shortening across the study area is 16.9% (5.8 km), and decreases south to north. Shortening estimates from the Upper Ecca Group increase from 4.3 km near Matjiesfontein in the west to 10.4 km near Prince Albert in the east. Three dimensional restorations of stratigraphic surfaces are consistent with these figures (17%), and allow paleogeographic and isopach thickness maps to be resorted to their configuration at the time of deposition. Structural restoration can be routinely employed in outcrop studies to improve the accuracy of dimensions (e.g. volumetrics) and reconstructions (e.g. sediment dispersal patterns) derived from ancient sedimentary systems. The workflow presented here will add value to exhumed basin analogues by presenting pre-kinematic configurations at the frontal margins of fold-thrust belts