Using mycobacterium cell wall fraction to decrease equine chorionic gonadotropin after abortion.

BACKGROUND: Equine embryonic loss following the development of endometrial cups delays return to cyclicity due to the production of equine chorionic gonadotropin (eCG). Natural degradation of endometrial cups coincides with an influx of immune cells at 100-120 days of gestation, but therapeutic stimulation of reduced eCG production has been relatively unsuccessful. Recently, we observed an increase in pro-inflammatory cytokine production following the use of the immunostimulant mycobacterium cell wall fraction (MCWF). OBJECTIVES: To evaluate the efficacy of hysteroscopic-guided injection of MCWF on the accelerated decline of eCG secretion. STUDY DESIGN: In vivo experiment. METHODS: Mares were pharmacologically aborted at 40-45 days of gestation, and then divided into groups: MCWF-treated (6 mg MCWF suspended in 20 mL LRS; n = 10) and Control (20 mL LRS; n = 6). Five days after abortion, hysteroscopic-guided injection of endometrial cups was performed, with 1 mL of volume placed into each visible endometrial cup. This was repeated 7 days later. Trans-rectal ultrasonography was performed to monitor ovarian activity, and serum was obtained to assess eCG and cytokine concentrations. RESULTS: Concentrations of eCG decreased in the MCWF-treated group (p < 0.01) with a significant suppression noted as early as 14 days after onset of treatment and remained suppressed for the duration of the study. This coincided with an increase in peripheral IFN-γ (p < 0.01) and IL-1ß (p < 0.01) concentrations. Eight out of ten MCWF-treated mares (80%) developed pre-ovulatory follicles, in comparison to 2/6 controls (33%). A pre-ovulatory follicle was noted 23 ± 4 days after onset of treatment. MAIN LIMITATIONS: No pregnancy data was obtained following treatment. CONCLUSIONS: This is the first report of a treatment for the accelerated reduction of eCG following abortion. Stimulation of this process allowed mares to develop a pre-ovulatory follicle within a month of MCWF treatment onset, granting repeat attempts at breeding within the confines of a single breeding season.

FAZ assembly in bloodstream form Trypanosoma brucei requires kinesin KIN-E.

Trypanosoma brucei, the causative agent of African sleeping sickness, uses its flagellum for movement, cell division, and signaling. The flagellum is anchored to the cell body membrane via the flagellum attachment zone (FAZ), a complex of proteins, filaments, and microtubules that spans two membranes with elements on both flagellum and cell body sides. How FAZ components are carried into place to form this complex is poorly understood. Here, we show that the trypanosome-specific kinesin KIN-E is required for building the FAZ in bloodstream-form parasites. KIN-E is localized along the flagellum with a concentration at its distal tip. Depletion of KIN-E by RNAi rapidly inhibits flagellum attachment and leads to cell death. A detailed analysis reveals that KIN-E depletion phenotypes include failure in cytokinesis completion, kinetoplast DNA missegregation, and transport vesicle accumulation. Together with previously published results in procyclic form parasites, these data suggest KIN-E plays a critical role in FAZ assembly in T. brucei.

Social complexity, life-history and lineage influence the molecular basis of castes in vespid wasps.

A key mechanistic hypothesis for the evolution of division of labour in social insects is that a shared set of genes co-opted from a common solitary ancestral ground plan (a genetic toolkit for sociality) regulates caste differentiation across levels of social complexity. Using brain transcriptome data from nine species of vespid wasps, we test for overlap in differentially expressed caste genes and use machine learning models to predict castes using different gene sets. We find evidence of a shared genetic toolkit across species representing different levels of social complexity. We also find evidence of additional fine-scale differences in predictive gene sets, functional enrichment and rates of gene evolution that are related to level of social complexity, lineage and of colony founding. These results suggest that the concept of a shared genetic toolkit for sociality may be too simplistic to fully describe the process of the major transition to sociality.

Model for Teaching the Helping Babies Survive Course Series and Advanced Life Support in Obstetrics Workshops to Rural Skilled Birth Attendants in Uganda.

Background: Uganda, like much of Sub-Saharan Africa and other underserved regions continues to face the challenge of high neonatal and maternal mortality. The Helping Babies Survive (HBS) course and the Advanced Life Support in Obstetrics (ALSO) provide hands on education to train providers in key life-saving interventions. A uterine balloon tamponade (UBT) procedure can be life-saving in the event of uterine bleeding. The purpose of this implementation research is to gain more insight into the effectiveness of a tailored down 5-day combined HBS-ALSO-UBT course. In this study, we found that a tailored down 5-day combined HBS-ALSO-UBT could be performed with significantly improved self-assessment in diagnosing and managing a wide range of peripartum conditions.

Identification of Phytotoxic Levels of Copper and Nickel in Commercial Organic Soil Amendments Recycled from Poultry Farms and Municipal Wastes.

Commercial-scale recycling of agricultural and municipal wastes into organic soil amendments facilitates safe disposal of waste and reduces environmental contamination. However, phytotoxicity of commercial organic amendments to crops is a major concern to farmers. Consistent with this, commercial chicken manure and Milorganite (recycled from municipal waste) were found to be phytotoxic. Chicken manure aqueous extract contains 10.8 ppm Cu and 0.7 ppm Ni. The level of Cu and Ni in Milorganite is lower. The current study identified an aqueous solution containing 5 ppm Cu, lower than in chicken manure aqueous extract, was highly phytotoxic to mustard seeds germination. Therefore, phytotoxicity of chicken manure is in part due to Cu. An aqueous solution containing 1 ppm Ni was not phytotoxic; whereas 0.125 ppm Ni was phytotoxic when 62.5 ppm Na, which is nontoxic, was added to the solution. Therefore, synergistic effects of chemicals in the organic amendments may induce phytotoxicity.

Trypanosomes have divergent kinesin-2 proteins that function differentially in flagellum biosynthesis and cell viability.

Trypanosoma brucei, the causative agent of African sleeping sickness, has a flagellum that is crucial for motility, pathogenicity, and viability. In most eukaryotes, the intraflagellar transport (IFT) machinery drives flagellum biogenesis, and anterograde IFT requires kinesin-2 motor proteins. In this study, we investigated the function of the two T. brucei kinesin-2 proteins, TbKin2a and TbKin2b, in bloodstream form trypanosomes. We found that, compared to kinesin-2 proteins across other phyla, TbKin2a and TbKin2b show greater variation in neck, stalk and tail domain sequences. Both kinesins contributed additively to flagellar lengthening. Silencing TbKin2a inhibited cell proliferation, cytokinesis and motility, whereas silencing TbKin2b did not. TbKin2a was localized on the flagellum and colocalized with IFT components near the basal body, consistent with it performing a role in IFT. TbKin2a was also detected on the flagellar attachment zone, a specialized structure that connects the flagellum to the cell body. Our results indicate that kinesin-2 proteins in trypanosomes play conserved roles in flagellar biosynthesis and exhibit a specialized localization, emphasizing the evolutionary flexibility of motor protein function in an organism with a large complement of kinesins.

Addressing the corona virus pandemic: will a novel filtered eye mask help?

OBJECTIVE: Non-hermetically sealed eye protection does not fully protect the eyes from airborne particles. Hermetically sealed eye protection fully protects the eyes from particles, but tends to fog up, rendering it unusable. This study aimed to build and test a filtered eye mask (FEM) to protect the eyes from airborne particles, while being usable without excessive fog build up. METHODS: The steps performed to build the FEM were described. A hermetically-sealed standard eye mask (SEM) and an FEM were examined at 1-minute, 5-minute and 60-minute periods for performance metrics relating to fog. RESULTS: The SEM showed minimal fog at 1minute, lots of fog at 5minutes and was dripping with condensation at 60minutes. The FEM was clear at 1minute, 5minutes and showed minimal fog at 60minutes. CONCLUSION: An FEM may play an important role in preventing novel coronavirus (COVID-19) exposure by protecting the eyes from airborne particles and preventing fog, rendering it usable. Further research is strongly recommended.

Movement Demands of an Elite Cricket Team During the Big Bash League in Australia.

There is a lack of research on the movement patterns within Twenty20 (T20) cricket, thus the purpose of this study was to investigate the movement demands placed on elite T20 cricket players playing in The Big Bash League, in Australia, in the 2017/2018 season. Player positional movements were determined from the time motion data obtained from a portable 10 Hz global positioning (GPS) unit. Overall, all the players covered between 1.77km and 6.54km in a time ranging between 40.4 minutes and 96.5 minutes. Fast bowlers covered a mean distance of 6.5 (±0.5) km, batsmen 1.7 (±1.2) km and fielders 5.9 (±0.9) km. This is the first study that has looked at the movement demands of players in The Big Bash League and found that bowlers have the highest movement demands followed by fielding. With that, arguably, more attention needs to be devoted to bowling and particularly fielding which is often not prioritized. However, overall demands of T20 cricketers have decreased. Cricketers and coaches need to ensure that they adapt training to ensure that their players are physically prepared for the associated demands.

Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares.

Pregnancy loss during the normal lifespan of endometrial cups (∼37-120-150 days of gestation) may affect a mare's ability to conceive again in the same breeding season, as equine chorionic gonadotropin (eCG) secretion by retained endometrial cups can lead to abnormal ovulations and follicular growth. While intrauterine kerosene infusion has anecdotally been proposed as a treatment for endometrial cup retention, there are no controlled studies evaluating kerosene's ability to enhance endometrial cup regression following abortion. The objectives of this study were to assess uterine response, systemic side effects, and efficacy of intrauterine kerosene infusions after abortion. We hypothesized that kerosene infusions would hasten regression of endometrial cups without detrimental effects on the endometrium and the mare's general health. Twelve light-breed mares were enrolled in the study after an experimentally induced abortion with cloprostenol (n = 12) by 60 ± 2 days of gestation. Mares were randomly allocated to receive an intrauterine infusion with 500 mL of kerosene (n = 6) or 500 mL saline (n = 6) on days 21 and 35 after pregnancy termination. Uterine biopsies were collected at days 7, 21, 35, and 49 post-abortion to evaluate the degree of endometrial fibrosis with Picrosirius Red Stain and to be graded according to the Kenney & Doig 1986 classification. Furthermore, histomorphometry analysis of the endometrium lining, glandular epithelium and glandular density was performed. Endometrial lymphocyte B CD20+, lymphocyte T CD3+, and macrophage IBA-1+ cell populations were characterized by immunohistochemistry. Physical examinations, blood cell counts, and serum biochemistry were performed before, and for 2 days after each uterine infusion. Serum samples were collected for assessment of eCG concentrations. Continuous data were analyzed with MIXED procedure with repeated measures in SAS, categorical data with LOGISTIC procedure of SAS. Significance was set at p < 0.05. Kerosene infusion did not affect complete blood cell counts, serum chemistry parameters, or physical examinations. Concentrations of eCG decreased over time (p < 0.001), but there were no differences between groups or time by group interactions (p = 0.72). Histological evaluation of the uterus showed no signs of increased fibrosis or degeneration in the treatment group. In conclusion, while kerosene infusions did not appear to have detrimental effects on mare health, our findings suggest that the use of kerosene in the uterus does not enhance the regression of endometrial cups by 49 days post-abortion.

Cryo-EM Structure (4.5-Å) of Yeast Kinesin-5-Microtubule Complex Reveals a Distinct Binding Footprint and Mechanism of Drug Resistance.

Kinesin-5s are microtubule-dependent motors that drive spindle pole separation during mitosis. We used cryo-electron microscopy to determine the 4.5-Å resolution structure of the motor domain of the fission yeast kinesin-5 Cut7 bound to fission yeast microtubules and explored the topology of the motor-microtubule interface and the susceptibility of the complex to drug binding. Despite their non-canonical architecture and mechanochemistry, Schizosaccharomyces pombe microtubules were stabilized by epothilone at the taxane binding pocket. The overall Cut7 footprint on the S. pombe microtubule surface is altered compared to mammalian tubulin microtubules because of their different polymer architectures. However, the core motor-microtubule interaction is tightly conserved, reflected in similar Cut7 ATPase activities on each microtubule type. AMPPNP-bound Cut7 adopts a kinesin-conserved ATP-like conformation including cover neck bundle formation. However, the Cut7 ATPase is not blocked by a mammalian-specific kinesin-5 inhibitor, consistent with the non-conserved sequence and structure of its loop5 insertion.

Photoelectrochemical Characterisation on Surface-Inverted Black Silicon Photocathodes by Using Platinum/Palladium Co-catalysts for Solar-to-Hydrogen Conversion.

Black silicon (bSi) has recently captured research attention in photoelectrochemical (PEC) solar-to-hydrogen (STH) conversion devices. Because nanostructuring of silicon retains the photovoltaic attributes of the material, it also provides a range of excellent physicochemical properties, such as a vast active-site-rich electrochemical interface, owing to a high aspect ratio, and important light-scattering attributes, which significantly improve photoconversion. One method to gain control over p-type bSi interface energetics is surface inversion of the p-type interface by phosphorus doping to introduce a shallow n+ -emitter layer, which provides a thin p-n junction at the interface of the nanostructures. Although this concept has been suggested in the literature, it has not been demonstrated experimentally for a platinum/palladium co-catalysed bSi photocathode device for STH conversion. Herein, preliminary investigations and proof-of-concept studies are reported for the fabrication and PEC characterisation of surface-inverted p-type bSi photocathodes prepared by wet chemical etching. The PEC tests on p-bSi|n+ photocathodes show that, for both metal nanoparticles (Pt and Pd), the catalytic activity for proton conversion is increased; this is evident from an anodic shift in the onset potentials shifts to 0.24 and 0.29 V and an increase in photocurrent by 9 and 13.8 mA cm-2 , respectively, at 0 V versus a reversible hydrogen electrode, as a result of introducing the emitter layer.

Nucleotide- and Mal3-dependent changes in fission yeast microtubules suggest a structural plasticity view of dynamics.

Using cryo-electron microscopy, we characterize the architecture of microtubules assembled from Schizosaccharomyces pombe tubulin, in the presence and absence of their regulatory partner Mal3. Cryo-electron tomography reveals that microtubules assembled from S. pombe tubulin have predominantly B-lattice interprotofilament contacts, with protofilaments skewed around the microtubule axis. Copolymerization with Mal3 favors 13 protofilament microtubules with reduced protofilament skew, indicating that Mal3 adjusts interprotofilament interfaces. A 4.6-Å resolution structure of microtubule-bound Mal3 shows that Mal3 makes a distinctive footprint on the S. pombe microtubule lattice and that unlike mammalian microtubules, S. pombe microtubules do not show the longitudinal lattice compaction associated with EB protein binding and GTP hydrolysis. Our results firmly support a structural plasticity view of microtubule dynamics in which microtubule lattice conformation is sensitive to a variety of effectors and differently so for different tubulins.

A mitochondrial therapeutic reverses visual decline in mouse models of diabetes.

Diabetic retinopathy is characterized by progressive vision loss and the advancement of retinal micoraneurysms, edema and angiogenesis. Unfortunately, managing glycemia or targeting vascular complications with anti-vascular endothelial growth factor agents has shown only limited efficacy in treating the deterioration of vision in diabetic retinopathy. In light of growing evidence that mitochondrial dysfunction is an independent pathophysiology of diabetes and diabetic retinopathy, we investigated whether selectively targeting and improving mitochondrial dysfunction is a viable treatment for visual decline in diabetes. Measures of spatial visual behavior, blood glucose, bodyweight and optical clarity were made in mouse models of diabetes. Treatment groups were administered MTP-131, a water-soluble tetrapeptide that selectively targets mitochondrial cardiolipin and promotes efficient electron transfer, either systemically or in eye drops. Progressive visual decline emerged in untreated animals before the overt symptoms of metabolic and ophthalmic abnormalities were manifest, but with time, visual dysfunction was accompanied by compromised glucose clearance, and elevated blood glucose and bodyweight. MTP-131 treatment reversed the visual decline without improving glycemic control or reducing bodyweight. These data provide evidence that visuomotor decline is an early complication of diabetes. They also indicate that selectively treating mitochondrial dysfunction with MTP-131 has the potential to remediate the visual dysfunction and to complement existing treatments for diabetic retinopathy.

Photoreceptor regulation of spatial visual behavior.

PURPOSE: To better understand how photoreceptors and their circuits support luminance-dependent spatial visual behavior. METHODS: Grating thresholds for optokinetic tracking were measured under defined luminance conditions in mice with genetic alterations of photoreceptor activity. RESULTS: The luminance conditions that enable cone- and rod-mediated behavior, and the luminance range over which rod and cone functions overlap, were characterized. The AII amacrine pathway was found to support low-resolution and high-contrast function, with the rod-cone pathway supporting high-resolution and low-contrast function. Rods alone were also shown to be capable of driving cone-like spatial visual function, but only when cones were genetically maintained in a physiological dark state. CONCLUSIONS: The study defined how luminance signals drive rod- and cone-mediated spatial visual behavior and revealed new and unexpected contributions for rods that depend on an interaction between cone and rod systems.

Aspiration and injection of the knee joint: approach portal.

Aspiration and injection of the knee joint is a commonly performed medical procedure. Injection of corticosteroid for the treatment of osteoarthritis is the most common reason for knee joint injection, and is performed as an office procedure. Debate exists among practitioners as to the 'best' approach portal for knee injection. This paper examines the various approach portals for injection and/or aspiration of the knee joint, as well as the accuracy of each approach. Searches were made of electronic databases, and appropriate papers were identified and hand-searched. Although there is some evidence that particular approach portals may be more efficacious in the presence of specific knee joint pathologies, generally, in experienced hands, it is of no clinical consequence as to which approach portal is utilised for aspiration or injection of the knee joint. No approach portal is 100% accurate, and the accuracy of injection of the knee joint may be enhanced by the use of techniques such as ultrasound. Practitioners are reminded that they should continuously refine and practice their preferred technique. Knee joint aspiration and injection is a common, simple, and generally safe office procedure.

Injection of a chemical castration agent, zinc gluconate, into the testes of cats results in the impairment of spermatogenesis: a potentially irreversible contraceptive approach for this species?

Male sterilization by chemical agents is a nonsurgical contraceptive approach designed to induce azoospermia and, therefore, infertility. Intratesticular injection of zinc gluconate for sterilization of dogs has been described, but its use in cats remains limited. The objective of the present study was to evaluate, by light and transmission electron microscopy, the efficacy of a single intratesticular injection of a zinc gluconate solution (Testoblock) as a sterilant for male cats. Twelve sexually mature mixed breed cats were allocated at random into two groups (control = 6; treated = 6) and given a single injection into each testis of either isotonic saline or zinc gluconate, respectively. Histopathologic and ultrastructural evaluation was assessed at 120 days postinjection. Histopathologic changes were not detected in the testes from the control group. However, histologic evaluation of the treated group revealed atrophic and dilated seminiferous tubules, a decrease in the number of germ cells, and incomplete spermatogenesis. Sertoli cells had various degrees of cytoplasmic vacuolization. Intertubular tissue revealed active fibroblasts, collagen deposition, and inflammatory cells. The diameter of seminiferous tubules, epithelial height and tubular area were reduced (P < 0.05) in the treated group compared with controls. Azoospermia occurred in 8 of the 11 treated cats (73%). Ultrastructural evaluation of Leydig cells revealed loss of nuclear chromatin, increased smooth endoplasmatic reticulum, and mitochondria degeneration. Intratesticular injection of zinc gluconate solution impaired spermatogenesis in cats and has great potential as a permanent sterilant in this species.