Half Of Animals Research Articles

Сравнение радиопротекторных свойств рибоксина (инозина) и индралина при профилактическом введении в дозировках 100 мг/кг по критерию выживаемости облучённых мышей

Relevance: Due to the high chemical toxicity of all known effective radioprotectors, studies of the radioprotective properties of safer drugs are very relevant. A sufficient number of works are devoted to the radioprotective properties of ribonucleoside riboxin (inosine). However, studies comparing the direct radioprotective properties of riboxin and a recognized radioprotector, for example, indralin, using a survival test in irradiated animals have not yet been carried out. Purpose: Conduct a comparative assessment of the radioprotective properties of riboxin and indralin using a survival test in mice exposed to external X-ray radiation. Material and methods: The experiment was carried out on 200 male ICR (CD-1) mice of the SPF category in duplicate. In each experiment, the animals were divided into the following groups, separated by body weight, 10 animals each: vivar control, not exposed to drugs and radiation, radiation control, with preliminary intraperitoneal administration of sterile water and exposed to external X-ray radiation in doses of 6.0, 6.5 and 6.75 Gy, experimental groups exposed to irradiation in the indicated doses with preliminary intraperitoneal administration of riboxin at a dosage of 100 ml/kg body weight or indralin at a dosage of 100 ml/kg. Survival was assessed for 30 days after irradiation. The dose change factor was determined using probit analysis as the ratio of the radiation dose causing the death of half of the irradiated animals that received the drug to the radiation dose causing the death of half of the irradiated animals without administration of the drug. Results: The use of indralin before X-ray irradiation in doses of 6.0 Gy, 6.5 Gy and 6.75 Gy led to a statistically significant increase in the survival of animals compared to the group receiving Riboxin and control irradiation (р<0,05, log-rank test). Using equations derived from Phinney probit analysis, LD50 doses were calculated for indralin and riboxin, from which dose change factors were calculated to be 1.8 and 1.07, respectively. Conclusion: Since riboxin has not demonstrated radioprotective properties, its preventive use with intraperitoneal administration under the conditions described in this paper, for leveling the effects of radiation can be considered ineffective.

Kisspeptin-10 Improves Testicular Redox Status but Does Not Alter the Unfolded Protein Response (UPR) That Is Downregulated by Hypothyroidism in a Rat Model.

Hypothyroidism compromises the testicular redox status and is associated with reduced sperm quality and infertility in men. In this regard, studies have demonstrated the antioxidant potential of kisspeptin in reproductive and metabolic diseases. In this study, we evaluate the effects of kisspeptin-10 (Kp10) on the testicular redox, as well as mediators of the unfolded protein response (UPR) in adult rats with hypothyroidism. Adult male Wistar rats were randomly separated into the Control (n = 15), Hypo (n = 13) and Hypo + Kp10 (n = 14) groups, and hypothyroidism was induced with 6-propyl-2-thiouracil (PTU) for three months. In the last month, half of the hypothyroid animals received Kp10. Testis samples were collected for enzymatic, immunohistochemical and/or gene evaluation of mediators of oxidative stress (TBARs, lipid hydroperoxides (LOOH), ROS, peroxynitrite, SOD, CAT and GPX), endoplasmic reticulum stress (GRP78, ATF6, PERK, CHOP, HO-1 and sXBP1) and antiapoptocytes (BCL-2). Hypothyroidism increased apoptosis index, TBARS and LOOH concentrations, and reduced testicular gene expression of Sod1, Sod2 and Gpx1, as well as the expression of Grp78, Atf6, Ho1 and Chop. Treatment with Kp10, in turn, reduced testicular apoptosis and the production of peroxynitrite, while increased SOD1 and GPX ½ expression, and enzymatic activity of CAT, but did not affect the lower expression of UPR mediators caused by hypothyroidism. This study demonstrated that hypothyroidism causes oxidative stress and dysregulated the UPR pathway in rat testes and that, although Kp10 does not influence the low expression of UPR mediators, it improves the testicular redox status, configuring it as an important antioxidant factor in situations of thyroid dysfunction.

Phytomineral complexes as elements of functional nutrition to enhance the protective function of the lymphatic system in aging

Background: The problem of aging remains relevant in the modern world, with a notable connection between aging and nutrition. It is important to find a way to slow down aging. Integration lymphology with functional food theory (or phytodietics) is the most promising idea. Detection of lymphotropic properties in bioactive food ingredients will increase the lymphatic system activity in aging. Our task is to improve the protective status of lymph nodes by taking a phytomineral product in aging. Objective: The research purpose is to provide evidence of the use of a plant-mineral complex to activate the lymph system (lymph nodes) in aging. Participants in the experiment are 160 Wistar rats divided into groups of old and young animals. Half of the aging animals took an additional food plant-mineral complex "IQdetoxSORB". The basis of the food complex is physiologically acting substances such as flavonoids, trace elements, food fibers, and others.Methods: Different methods have been applied to study systems forming components of lymph nodes. The system of lymph node compartments was examined by a histological method. The liquid component of the organ is evaluated by a thermogravimetric method. Lymph node trace elements (Mn, Fe, Cu, Zn, Se) were determined by X-ray fluorescence analysis using synchrotron radiation (RFA SR). The StatPlus Pro license program and statistical analysis of dynamic series were used to process digital data.Results: We first presented data on the effect of functional nutrition on the lymphatic system and proved the lymphotropic properties of the plant-mineral complex. The bioactive ingredients of the complex (flavonoids, trace elements, food fibers) are useful and influence the lymphatic system, improving health in old age. The increase in lymphatic system activity occurs through the implementation of a lymphosanation mechanism due to the stimulating and protective effects of plant ingredients. Intake of the phytomineral complex accelerates lymph flow, modifies lymph node compartments, and restores trace element balance. The positive effect depends on the formation of an association of bioelements with lymphoid compartments. A plant-based diet corrects and increases the protective status of lymph nodes. The result is a slowdown in age-induced changes in peripheral lymphoid organs. Conclusion: As an element of functional nutrition, the plant-mineral complex contains bioactive ingredients - flavonoids, trace elements, and food fibers. These bioactive substances alter the existing status of the lymphatic system, increasing protective potential. Increased activity of the lymph system (lymph nodes) is an argument for slowing aging and preventing age-associated pathology. The information presented is the basis for recommending plant-mineral complexes for wellness programs of anti-aging.Keywords:lymphatic system; lymph nodes; compartments; trace elements; gerontology, phytotherapy, nutriciology.

Compound Loss of Dnmt3a and Kmt2c in Myeloid Malignancies

Monosomy 7, or loss of 7q (-7/del(7q)), is one the most common chromosomal anomalies in myelodysplastic syndromes (MDS, ~10% of patients), a malignant disorder of the blood system with few treatment options. There are currently no targeted therapies available for -7/del(7q) MDS, and patients progress to a deadly acute myeloid leukemia (AML). The 7q segment is rich in genes that are implicated in MDS pathophysiology, including KMT2C encoding a histone H3 lysine 4 methyltransferase (depositing H3K4me1) that marks active gene enhancers during myeloid differentiation. Co-occurring DNA methyltransferase DNMT3A mutations are significantly overrepresented in -7/del(7q) MDS, especially in the absence of other chromosomal aberrations (23%). Individually, loss in DNMT3A activity in hematopoietic stem cells (HSCs) has shown bias towards self-renewal due to a failure to shut off stem-cell-related genes, whereas a loss of KMT2C leads to aberrant myelopoiesis. Generally, only one allele of KMT2C and DNMT3A is lost in MDS, suggesting that these haploinsufficient tumor suppressors may need to cooperate to drive disease. We hypothesized that disruption of Dnmt3a cooperated with loss of Kmt2c creating an aberrant epigenetic state that may accelerate development of MDS and progression to AML. To address this, we generated mice that lacked one copy of both Kmt2c and Dnmt3a in the bone marrow by conditional deletion with a hematopoietic-specific inducible Mx1-driven Cre recombinase. After inducing Cre-mediated recombination by poly(I:C) treatment creating Kmt2c(+/del):Dnmt3a(+/del) animals, the hematopoietic stem and progenitor compartment was subjected to comprehensive immunophenotypic and functional characterization. In serial competitive bone marrow repopulation assays, cells with combined heterozygous loss of both Dnmt3a and Kmt2c demonstrated increased competitive advantage in secondary transplants and retained ability to reconstitute tri-lineage hematopoiesis in tertiary transplants, unlike single gene deletion and wild-type controls. Consistently, immunophenotypic analysis of the bone marrow compartment 16 weeks after the third transplant demonstrated persistence of the primitive long-term hematopoietic stem cells (LT-HSCs) defined as Lineage -Sca1 +cKit + (LSK) CD150 +CD48 -, which were absent in other genotypes. Further, we found expansion of the LT-HSCs in aged 80-week old Kmt2c(+/del):Dnmt3a(+/del) animals that were also characterized by a gain in self-renewal as seen by their ability to serially replate in semisolid media (MethoCult GM3434). To test if combined loss of Kmt2c and Dnmt3a led to disease development, next we established a large cohort of animals non-competitively transplanted with Kmt2c(+/del):Dnmt3a(+/del), single gene deletion, and wild-type control bone marrow, and initiated a tumor watch by monthly monitoring the composition of their peripheral blood by CBCs and flow cytometry. Sixty weeks after transplantation, over a half of double knock-out animals developed myeloid disease characterized by neutrophilia and/or monocytosis accompanied by anemia, occasional thrombocytopenia, and splenomegaly most consistent with myelodysplasia. Immunophenotypically, we found expanded LSK populations (LT-HSCs specifically) in the bone marrow and evidence of extramedullary hematopoiesis in the spleen. At the same time, the majority of mice with single gene deletion and all wild-type controls remained healthy. Overall, our studies demonstrate that combined loss of Kmt2c and Dnmt3a results in the development of myelodysplastic disease with long latency through endowing the HSCs with enhanced self-renewal capacity. We are currently interrogating the epigenetic profiles and gene expression patterns resulting from combined disruption of Dnmt3a and Kmt2c aiming to identify gene targets and signaling networks for future therapeutic intervention. Better understanding of the molecular pathogenesis of the -7/del(7q) MDS with concurrent DNMT3A mutations is critical to develop more effective treatment approaches and to improve outcomes for MDS patients.

Kisspeptin treatment reverses high prolactin levels and improves gonadal function in hypothyroid male rats

We evaluated whether the administration of kisspeptin-10 (Kp10) is capable of restoring gonadal function in hypothyroid male rats. Hypothyroidism was induced with 6-propyl-2-thiouracil (PTU) for three months. In the last month, half of the hypothyroid animals were treated with Kp10. Hypothyroidism reduced testicular and sex gland mass, decreased the proliferation of the seminiferous epithelium, and compromised sperm morphology, motility, and vigor. A decrease in plasma LH and testosterone levels and an increase in prolactin secretion were observed in the hypothyroid rats. Hypothyroidism reduced Kiss1 and Kiss1r protein and gene expression and Star and Cyp11a1 mRNA levels in the testis. Furthermore, it reduced Lhb, Prl, and Drd2 and increased Tshb and Gnrhr expression in the pituitary. In the hypothalamus, hypothyroidism increased Pdyn and Kiss1r while reducing Gnrh1. Kp10 treatment in hypothyroid rats restored testicular and seminal vesicle morphology, improved sperm morphology and motility, reversed high prolactin levels, and increased LH and testosterone levels. In addition, Kp10 increased testicular expression of Kiss1, Kiss1r, Fshr, and Nr5a1 and pituitary Kiss1 expression. Our findings describe the inhibitory effects of hypothyroidism on the male gonadal axis and sperm quality and demonstrate that Kp10 treatment reverses high prolactin levels and improves gonadal function and sperm quality in hypothyroid rats.

Le fantastique dans les romans d’Ananda Devi

In the article, the author analyses the fantasy elements noticeable in some of Ananda Devi’s novels. In her first novels as well as in her recent ones, the writer resorts to solutions from the fantasy genre. In L’Arbre fouet, Moi, l’interdite, La Vie de Joséphin le Fou, Pagli, Soupir, Indian tango, Les Jours vivants, Manger l’autre and in Le Jour des caméléons, the Mauritian novelist presents mysterious incipits and open-ended, dreamlike endings, introduces hybridised, often insane characters half animal half human, describes metamorphoses and transformations of man into animal, mentions imaginary sobriquets, ghosts. The aim of the analysis is also to try to explain for what purpose the writer uses fantasy in her work. In the analysis, the author also draws on the research of T. Todorov, P.-G. Castex, L. Vax and V. Tritter.

Positive Effects of the Nutraceutical Association of Lycopene and Selenium in Experimental Varicocele.

Many natural substances commonly found in healthy diets have been studied for their potential to reduce male infertility associated with varicocele. A positive role of selenium (Se) or lycopene alone was demonstrated in experimental varicocele, while no data are available on their association. One group of male Sprague-Dawley rats was sham operated and daily treated with Se (3 mg/kg, i.p.), lycopene (1 mg/kg, i.p.), or their association. A second group underwent surgery to induce varicocele. Sham and half of the varicocele animals were sacrificed after twenty-eight days, while the residual animals were treated for one more month and then sacrificed. In varicocele animals, testosterone levels and testes weight were reduced, Hypoxia Inducible Factor-1α (HIF-1α) expression was absent in the tubules and increased in Leydig cells, caspare-3 was increased, seminiferous epithelium showed evident structural changes, and many apoptotic germ cells were demonstrated with TUNEL assay. The treatment with lycopene or Se alone significantly increased testis weight and testosterone levels, reduced apoptosis and caspase-3 expression, improved the tubular organization, decreased HIF-1α positivity of Leydig cells, and restored its tubular positivity. Lycopene or Se association showed a better influence on all biochemical and morphological parameters. Therefore, the nutraceutical association of lycopene plus Se might be considered a possible therapeutic tool, together with surgery, in the treatment of male infertility. However, long-term experimental and clinical studies are necessary to evaluate sperm quantity and quality.

CUX1-related neurodevelopmental disorder: deep insights into phenotype-genotype spectrum and underlying pathology

Heterozygous, pathogenic CUX1 variants are associated with global developmental delay or intellectual disability. This study delineates the clinical presentation in an extended cohort and investigates the molecular mechanism underlying the disorder in a Cux1+/− mouse model. Through international collaboration, we assembled the phenotypic and molecular information for 34 individuals (23 unpublished individuals). We analyze brain CUX1 expression and susceptibility to epilepsy in Cux1+/− mice. We describe 34 individuals, from which 30 were unrelated, with 26 different null and four missense variants. The leading symptoms were mild to moderate delayed speech and motor development and borderline to moderate intellectual disability. Additional symptoms were muscular hypotonia, seizures, joint laxity, and abnormalities of the forehead. In Cux1+/− mice, we found delayed growth, histologically normal brains, and increased susceptibility to seizures. In Cux1+/− brains, the expression of Cux1 transcripts was half of WT animals. Expression of CUX1 proteins was reduced, although in early postnatal animals significantly more than in adults. In summary, disease-causing CUX1 variants result in a non-syndromic phenotype of developmental delay and intellectual disability. In some individuals, this phenotype ameliorates with age, resulting in a clinical catch-up and normal IQ in adulthood. The post-transcriptional balance of CUX1 expression in the heterozygous brain at late developmental stages appears important for this favorable clinical course.

Francisella tularensis PCR detection in Cape hares (Lepus capensis) and wild rabbits (Oryctolagus cuniculus) in Algeria

Tularemia is a zoonosis caused by the bacterium Francisella tularensis. Leporids are primary sources of human infections in the northern hemisphere. Africa is classically considered free of tularemia, but recent data indicate that this dogma might be wrong. We assessed the presence of this disease in wild leporids in Algeria. Between 2014 and 2018, we collected 74 leporids carcasses from spontaneously dead or hunted animals. Francisella tularensis DNA was detected by specific real-time PCR tests in 7/36 (19.44%) Cape hares (Lepus capensis) and 5/38 (13.15%) wild rabbits (Oryctolagus cuniculus). Known tularemia arthropod vectors infested half of the PCR-positive animals. At necropsy, F. tularensis-infected animals presented with an enlarged spleen (n = 12), enlarged adrenal glands (12), liver discoloration (12), hemorrhages (11), and pneumonia (11). Immunohistological examination of liver tissue from one animal was compatible with the presence of F. tularensis. Our study demonstrates the existence of tularemia in lagomorphs in Algeria. It should encourage investigations to detect this disease among the human population of this country.

ELAVL1/HuR regulates both endo‐ and exogenous neuroprotection in retinal ganglion cells

AbstractPurpose: To evaluate the impact of HuR gene silencing on the ratio of age‐related degeneration of Retinal Ganglion Cells (RGC), which potentially describes the efficiency of endogenous neuroprotection mechanisms, as well as to assess the exogenous neuroprotection capacity of HuR‐ silenced RGC in rat glaucoma model treated with Metallothionein.Methods: Thirty‐five eight‐week‐old male Long Evans rats were divided into two groups: experimental and control. Animals from experimental group received intravitreal injection of AAV‐shRNA‐HuR. Control group received injection of AAV‐shRNA scramble control. Animals from both groups were sacrificed in 3 different time points: 2, 4 and 6 months after injection. Both healthy and treated retinas from each animal were collected and processed as whole mounts for immunostainings and RGC count. For the second trial, animals were divided into groups and transfected with virus particles as described above. To induce chronic glaucoma model, 8 weeks after AAV injection, unilateral episcleral vein cauterization was performed. Half of animals from each group received intravitreal injection of Metallothionein (MT), the other group received PBS injection. During the experiment IOP was monitored. Both healthy and treated retinas from each animal were collected and processed as whole mounts for immunostainings and RGC count.Results: RGC count (per visual field under 20× magnification) was 310 ± 31, 296 ± 25, 189 ± 41 in experimental group and 399 ± 51, 395 ± 49, 390 ± 23 in control respectively for 2, 4 and 6 months after injection (Kaplan‐Mayer trend rank p < 0.0001). Loss of RGC in central retina was 33.7% in animals from shRNA‐MT‐treated glaucoma and 11.4% in shRNA control‐MT‐treated glaucoma (p < 0.05). In peripheral part of the retina the loss was 37.4% in animals from shRNA‐MT‐treated glaucoma and 11.5% in shRNA control‐MT‐treated glaucoma (p < 0.01).Conclusions: Silencing of hur gene enhanced age‐related loss of RGC which translates into impaired endogenous neuroprotection pathways, moreover the absence of HuR critically limits neuroprotective activity of exogenously delivered Metallothionein.

Flock Nocturnal Activity: Is There a Rotative Guard?

Animal activity during the night period is of enormous importance, since it represents approximately half of animals’ lives, and monitoring it during this period makes it possible to detect problems related to well-being and safety, and allows us to infer energy expenditure on the basis of their activity level. The present study analyzes a sheep activity dataset created during the night period to validate non-invasive techniques of monitoring that can be used to infer energy expenditure at night and to detect abnormal nocturnal activity. The study allowed us to detect cyclic changes in activity during the night period, which is composed of inactive and active periods, and to identify sheep lying positions. The analysis of the joint activity of the flock allowed us to perceive a time lag in the rest cycles, which consisted of periods of activity of ewes undone between elements of the flock. Although it does not allow us to identify the components of the period of inactivity, since the method used does not monitor brain activity, the results allow us to confirm the cyclical character of the nocturnal activity of sheep that has been reported in the literature, as well as their typical posture when lying down. Although this is an exploratory application with a very small number of animals, the similarity between the results obtained and the results documented in the existing literature, which have mostly been obtained using invasive methods, is encouraging, and suggests it is possible to rely on activity monitoring processes based on inertial sensors.

Angiogenic and Inflammatory Alterations of Endometriotic Lesions in a Transgenic Animal Experimental Model With Loss of Expression of PPAR-Alpha Receptors

Peroxisome proliferator-activated receptors (PPARs) have been proposed as a medical treatment against endometriosis in preclinical and clinical studies. Their effect seems to be triggered through the suppression of angiogenesis. In the present study, we used a transgenic animal model with a loss of expression of PPAR-alpha receptors to examine their effect on the course of surgically induced endometriotic lesions. Ten C57BL/6 mice that served as controls and 10 B6;129S4-PPARatm1Gonz/J t transgenic mice characterized by absolute loss of expression of PPAR-alpha receptors were used for induction of endometriosis with a previously described surgical technique. Five animals (50%) exhibited abundant endometriotic crypts in the control group whereas only one (10%) animal in the transgenic experimental group had a similar pathological image. Neo-vascularization significantly differed among the two groups (p=0.034) favoring the control group as it was extremely limited in half of the PPAR-alpha null animals. The median inflammation score was 2.5 (1-4) in the P B6;129S4-PPARatm1Gonz/J group, whereas it was minimal, 1 (0-2), in the C57BL/6 group. However, these differences were not statistically significant (p=0.101). The fibroblastic activity was also very limited in the PPAR-alpha-deficient model, whereas animals belonging to the control group exhibited an intermediate increase of this index (p=0.022). Surgically induced endometriotic implants in animals with loss of expression of PPAR-alpha receptors exhibit significant differences in their pathology compared to lesions induced in control animals. This information suggests that PPAR-alpha receptors have a significant impact on the course of the disease, indicating that they may serve as potential targets for future medical therapies.

Controlled release vaccine implants for delivery of booster immunisations

Most current animal vaccine regimes involve a primary vaccination followed sometime later by a booster vaccination. This presents challenges when vaccinating difficult to access animals such as livestock. Mustering livestock to deliver a vaccine boost is costly and stressful for animals. Thus, we have produced a platform system that can be administered at the same time as the priming immunisation and delivers payload after an appropriate delay time to boost the immune response, without need for further handling of animals. A 30 × 2 mm osmotically triggered polymer implant device with burst-release characteristics delivered the booster dose of a tetanus vaccine. Blood samples were collected from an experimental group that received the priming vaccine and implant on day 0 and control group that received the initial vaccine (tetanus toxoid) and then a bolus dose 28 days later via subcutaneous injection. The two groups showed identical weight gain curves. T cell proliferation following in vitro stimulation with antigen was identical between the two groups at all time points. However, serum IgG antibody responses to the tetanus toxoid antigen were significantly higher in the control group at weeks 8 and 12. The implant capsules stayed at the site of implantation and at week 12 there was evidence of tissue integration. No local reactions at the implant site were observed, other than mild thickening of the skin in half of the experimental group animals and no other adverse health events were recorded in either group.

Pro-inflammatory response markers and neutrophil activity index of mice treated with butaphosphan and challenged with bacterial endotoxin

Butaphosphan, an organic source of phosphorus, potentiates humoral and cellular immune responses, however, it is still not clear how this occurs. We evaluate the effect of subcutaneous administration of injectable butaphosphan on pro-inflammatory markers and neutrophil activity index of mice challenged with bacterial lipopolysaccharide. Forty-two mice were randomly divided into three groups: Butaphosphan 50 (BUT50, n=14), that received injections of 50 mg/kg of butaphosphan; Butaphosphan 200 (BUT200, n=14), that received injections of 200 mg/kg of butaphosphan and Control (CTL, n=15), that received placebo. Subcutaneous injections were performed twice daily, for eight consecutive days. At the 8th day, half of animals per group were randomly selected to be challenged with bacterial lipopolysaccharide. Two hours after the challenge animals were euthanized, blood and spleen collected to analyze hematological parameters, circulating levels of interleukin 6 (IL-6), 1β (IL-1B) and myeloperoxidase (MPO) tissue activity. Red blood cell count was higher in butaphosphan treated groups. White blood cell count was lower in BUT200/LPS, as consequence of the decreased neutrophil count. Defense cells and IL-6 were elevated in BUT200 group. The use of butaphosphan ameliorate inflammatory markers, both in challenged and not challenged animals.

Outsourcing the dry season: Cattle ranchers' responses to weather shocks in the Brazilian Amazon

AbstractLivestock production plays a vital role in the global economy, yet little is known about how climate change will affect the industry. Studies of crop agriculture may not translate to livestock agriculture due to differences in the set of potential adaptation strategies. Ranchers' responses to changing weather and climate are particularly relevant in the Brazilian Amazon, where the dry season is increasing by as much as 0.6 days per year. A longer dry season increases production risks as animals may succumb to starvation, lack of water, or heat stress. I pair transaction data and weather shocks to show that ranchers strategically sold animals prior to the dry season, in response to two predictors of an extreme dry season: sporadic rainfall and high temperatures prior to the dry season. Ranchers sold animals for both fattening and slaughter in response to sporadic rainfall prior. Nearly half of animals initially sold for fattening were sold for slaughter 90 days later. In contrast, ranchers responded to high temperatures solely by selling for slaughter, suggesting that current on‐farm technologies are insufficient to prevent losses from heat stress. The overall supply increased in years with sporadic rainfall and high temperatures, but the supply fell by a greater amount the following year. My results suggest that farmers are currently not fully insulated from the effects of a severe dry season. As severe dry seasons become more frequent, the industry will likely increasingly rely on feedlots and need additional methods to cope with heat stress.

Experimental Rickettsia typhi Infection in Monodelphis domestica: Implications for Opossums as an Amplifying Host in the Suburban Cycle of Murine Typhus.

Murine typhus is an acute undifferentiated febrile illness caused by Rickettsia typhi. In the United States, its reemergence appears to be driven by a shift from the classic rat-rat flea cycle of transmission to one involving opossums (Didelphis virginiana) and cat fleas. Little is known of the ability of opossums to act as a reservoir and amplifying host for R. typhi. Here, we use Monodelphis domestica (the laboratory opossum) as a surrogate for D. virginiana. Opossums were inoculated via the intraperitoneal (IP) or intradermal (ID) route with 1 × 106 viable R. typhi. Blood and tissues were collected on days 6, 13, 20, and 27 or if moribund. Although one ID-infected opossum died, the remainder did not appear ill, whereas half of the IP-inoculated animals succumbed to infection. Rickettsemia was demonstrated in all animals through week 2 of infection and sporadically in weeks 3 and 4. Rickettsia typhi DNA was detected in all tissues, with most animals demonstrating the presence of bacteria into weeks 3 and 4. Histopathology and immunohistochemistry demonstrated typical findings of rickettsial infection. Akin to infection in rats, the demonstration of disseminated infection, typical inflammation, and prolonged rickettsemia with relatively few clinical effects (especially in the more natural route of ID inoculation) supports the potential of opossums to act as a competent mammalian reservoir and component of the zoonotic maintenance cycle of R. typhi. Understanding the dynamics of infection within opossums may have implications for the prevention and control of murine typhus.

Neuronal Nitric Oxide Synthase expression in brain from male spontaneously hypertensive rats. Implication in the control of cardiovascular function

Neuronal nitric oxide synthase (nNOS) in brain has been implicated in the regulation of sympathetic tone and control of cardiovascular functions, then in this study it was determined the nNOS expression and nitrites levels in brain from spontaneously hypertensive rats (SHR). Male Wistar Kyoto (WKY) and (SHR) strains at 4 months old were used, half of the experimental animals were administered with reserpine (R), (1 mg/kg of weight, for two weeks) and the others received vehicle (V) administration (dimethyl sulfoxide). Four experimental groups (WK+V, WKY+R, SH+V, SH+R) were followed and after cardiovascular parameters determination, brain was obtained. The expression of nNOS through western blot in frontal cortex‐FC, occipital cortex‐OC, cerebellum‐CB, medulla oblongata‐MO, spinal cord‐EC was determined and nitrites levels in tissue homogenate from the same brain regions were measured through Griess reaction. Heart rate (HR), systolic arterial blood pressure (SABP) and body weight, were analyzed. The results show that reserpine decrease significantly the body weight in g (WK+V 335.2±17.6 versus WK+R 280.4±9.87; SH+V 334.8±9.01 versus SH+R 269.3±8.65), the HR in beat/min (WK+V 302.2±11.6 versus WK+R 264±9.13; SH+V 354±17 versus SH+R 224±16), SABP in mmHg (WK+V: 165.8±4,964, versus WK+R 118.2±7.52; SH+V 215.4±5,074 versus SH+R 139.6±3.75), in both WKY and SHR (P<0.05 for all parameters). nNOS protein expression was high in WK+V in comparison with the SH+V, in FC and CB, whereas in the groups with reserpine administration the nNOS expression was higher in SH+R versus SH+V in FC, CB and MO. Tissue nitrites levels from different regions of the brain were reduced significantly in the SHR in comparison with WKY, particularly in FC (WK+V‐5.11 ±0.60 versus SH+V 3.53±0.53) and MO (WK+V 7.84±0.84 versus SH+V 3.24±0.48), however in the groups SHR+R a significantly increased of nitrites levels was observed in comparison with their own control, particularly in CB (SH+V 3.91±1.04 versus SH+R 7.53±0.83) and MO (SH+V 3.72±0.44 versus SH+R 5.8±0.39) (P<0.05 for all parameters). Increased nNOS expression in frontal cortex and cerebellum produces a significant decrease in the heart rate and systemic arterial blood pressure suggesting a key role of nNOS in the regulation of cardiovascular function in the SHR.

Partial exogastrulation due to apical-basal polarity of F-actin distribution disruption in sea urchin embryo by omeprazole.

Gastrulation is a universal process in the morphogenesis of many animal embryos. Although morphological and molecular events in gastrulation have been well studied, the mechanical driving forces and underlying regulatory mechanisms are not fully understood. Here, we investigated the gastrulation of embryos of a sea urchin, Hemicentrotus pulcherrimus, which involves the invagination of a single‐layered vegetal plate into the blastocoel. We observed that omeprazole, a proton pump inhibitor capable of perturbing the left–right asymmetry of sea urchin embryo, induced “partial exogastrulation” where the secondary invagination proceeds outward. During early gastrulation, intracellular apical–basal polarity of F‐actin distribution in vegetal half was higher than those in animal half, while omeprazole treatment disturbed the apical–basal polarity of F‐actin distribution in vegetal half. Furthermore, gastrulation stopped and even partial exogastrulation did not occur when F‐actin polymerization or degradation in whole embryo was partially inhibited via RhoA or YAP1 knockout. A mathematical model of the early gastrulation reproduced the shapes of both normal and exogastrulating embryos using cell‐dependent cytoskeletal features based on F‐actin. Additionally, such cell position‐dependent intracellular F‐actin distributions might be regulated by intracellular pH distributions. Therefore, apical–basal polarity of F‐actin distribution disrupted by omeprazole may induce the partial exogastrulation via anomalous secondary invagination.

Revealing the cells fated to form the cell mass in embryos of temnopleurid sea urchins

Larvae of temnopleurid sea urchins form a cell mass (CM) instead of an amniotic cavity on the left side at the early developmental stage for formation of the adult rudiment. However, the cell lineage and the mechanisms that form the CM are still unknown. We analyzed the potential to form a CM in partial embryos resulting from microsurgeries, using two temnopleurid species, Mespilia globulus (L.) and Temnopleurus toreumaticus (Leske). CM formation was completed 28-34 h after fertilization at 24°C, corresponding to the period from the late prism to the two-armed pluteus larval stages in both species. In the case of specimens dissected horizontally during the mesenchyme blastula to prism stages, the CM was formed in partial embryos containing enough of the an2 region, a part of the precursor cells that differentiate the ectoderm. The proportion of specimens with a CM was higher in T. toreumaticus than in M. globulus. Additionally, all larvae derived from half embryos obtained from dissection along the animal-vegetal axis at the mesenchyme blastula stage formed the CM. Transplantation of a stained animal or vegetal hemisphere at the 16-cell stage into a nonstained vegetal or animal embryo indicated that the CM derives from the animal half. Exogastrulae vegetalized by lithium chloride treatment did not form the CM. These results indicate that the CM formation is dependent not only on the an2 region but also on signals from the vegetal region after the mesenchyme blastula stage.

THE MORPHOLOGICAL CHARACTERISTICS OF THE OFFSPRING OF MALE RATS IRRADIATED WITH ELECTRONS AT A DOSE OF 2 GY AND TREATMENT BY PLATELET-RICH PLASMA

One of the most dangerous effects of radiation on health is reproductive dysfunction, often leading to infertility. Available studies on the effects of radiation on spermatogenesis and fertility have not considered the possibility of using restorative therapy. One of the modern methods of infertility treatment is the introduction of autologous blood plasma with an increased content of platelets and native growth factors. Currently, there are no studies on the study of offspring obtained by using this method of infertility treatment and the assessment of its teratogenicity. Purpose of the study: morphological analysis of offspring of male rats irradiated with electrons at a dose of 2 Gy and receiving platelet-rich plasma. Male Wistar rats (n=35) were once subjected to targeted ionizing radiation at a NOVAC-11 pulsed electron accelerator at a dose of 2 Gy. To half (n=15) of the irradiated animals were injected with platelet-rich plasma and insulin-like growth factor-1 for 12 weeks, after which the males of both groups were placed with intact females (n=30) to produce offspring. The state of the developing offspring was assessed by morphological analysis after the removal of pregnant females from the experiment on the 20th day of prenatal development. Pregnancy was observed only in females bred by control males and males treated with platelet-rich plasma. Classical histological methods and X-ray computed microtomography were used. As a result of the study, it was found that the weight and size of the offspring from the control and experimental groups did not have macroscopic differences; light microscopy did not reveal significant changes in their histological structure. Pathological changes in the development of the offspring of males of the experimental group who received platelet-rich plasma after electron irradiation with a dose of 2 Gy using computed microtomography were also not detected. The obtained data indicate about of effectiveness of the treatment of the infertility of irradiated male rats with platelet-rich plasma, as well as the absence of its teratogenic effects.