Survival Rate Of Animals Research Articles

Characteristics of structural and functional alterations following traumatic brain injury in neurons and glial cells of the sensorimotor cortex

Introduction and Aim: Traumatic brain injury (TBI) is regarded as a significant worldwide health issue and a significant contributor to mortality and disability. The objective of this study was to examine the parameters and nature of the regenerative and destructive processes that occur in the rat cerebral cortex depending on the degree and time of the TBI. Materials and Methods: The experiments were carried out on 24 sexless adult mice weighing 180–220 g. The primary group of mice had severe TBI. To assess the severity of the TBI model, histological data, animal survival rates, and motor and cognitive dysfunctions were examined. Both light and electron microscopy were used to study the animal brains in each group. Results: The areas of injury were filled with blood, and microscopic examination revealed that the foci of contusion had destroyed brain tissue in the form of tissue and blood vessel fragments. Most dystrophic neuronal changes in mice with severe TBI between 1 and 21 days after the injury were acute neuronal swelling, hydropic dystrophy of nerve cells with clear cytoplasmic vacuolization, localized and complete chromatolysis, hyperchromatism, and homogeneous cytoplasm. Conclusion: Brain tissue lesions develop in the early stages of a TBI through rapid necrotic cell death.

Famotidine Enhances Rifampicin Activity against Acinetobacter baumannii by Affecting OmpA.

The development of novel antibiotic adjuvants is imminent because of the frequent emergence of resistance in Gram-negative bacteria, which severely restricts the efficiency and longevity of commonly used clinical antibiotics. It is reported that famotidine, a clinical inhibitor of gastric acid secretion, enhances the antibacterial activity of rifamycin antibiotics, especially rifampicin, against Gram-negative bacteria and reverses drug resistance. Studies have shown that famotidine disrupts the cell membrane of Acinetobacter baumannii and inhibits the expression of the outer membrane protein ompA gene, while causing a dissipation of the plasma membrane potential, compensatively upregulating the pH gradient and ultimately increasing the accumulation of reactive oxygen species by leading to increased bacterial mortality. In addition, famotidine also inhibited the efflux pump activity and the biofilm formation of A. baumannii. In the Galleria mellonella and mouse infection models, the combination of famotidine and rifampicin increased the survival rate of infected animals and decreased the bacterial load in mouse organs. In conclusion, famotidine has the potential to be a novel rifampicin adjuvant, providing a new option for the treatment of clinical Gram-negative bacterial infections. IMPORTANCE In this study, famotidine was discovered for the first time to have potential as an antibiotic adjuvant, enhancing the antibacterial activity of rifamycin antibiotics against A. baumannii and overcoming the limitations of drug therapy. With the discovery of novel applications for the guanidine-containing medication famotidine, the viability of screening prospective antibiotic adjuvants from guanidine-based molecules was further explored. In addition, famotidine exerts activity by affecting the OmpA protein of the cell membrane, indicating that this protein might be used as a therapeutic drug target to treat A. baumannii infections.

Adjuvant Oligonucleotide Vaccine Increases Survival and Improves Lung Tissue Condition of B6.Cg-Tg (K18-ACE2)2 Transgenic Mice

The main problem in creating anti-coronavirus vaccines that target mainly proteins of the outer membrane of the virus is the rapid variability in the RNA genome of the pathogen that encodes these proteins. In addition, the introduction of technologies that can affordably and quickly produce flexible vaccine formulas that easily adapt to the emergence of new subtypes of SARS-CoV-2 is required. Universal adjuvant oligonucleotide vaccines based on conserved regions of the SARS-CoV-2 genome can take into account the dynamics of rapid changes in the virus genome, as well as be easily synthesized on automatic DNA synthesizers in large quantities in a short time. In this brief report, the effectiveness of four phosphorothioate constructs of the La-S-so-type adjuvant oligonucleotide vaccine is evaluated on B6.Cg-Tg (K18-ACE2)2 transgenic mice for the first time. In our primary trials, the oligonucleotide vaccine increased the survival rate of animals infected with SARS-CoV-2 and also reduced the destructive effects of the virus on the lung tissue of mice, activating both their innate and adaptive immunity. The obtained results show that the development of adjuvant oligonucleotide vaccine constructs of the La-S-so type is an affordable and efficient platform for the prevention of coronavirus infections, including those caused by SARS-CoV-2.

Broadly Effective ACE2 Decoy Proteins Protect Mice from Lethal SARS-CoV-2 Infection.

As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants have been causing increasingly serious drug resistance problem, development of broadly effective and hard-to-escape anti-SARS-CoV-2 agents is an urgent need. Here, we describe further development and characterization of two SARS-CoV-2 receptor decoy proteins, ACE2-Ig-95 and ACE2-Ig-105/106. We found that both proteins had potent and robust in vitro neutralization activities against diverse SARS-CoV-2 variants, including BQ.1 and XBB.1, that are resistant to most clinically used monoclonal antibodies. In a stringent lethal SARS-CoV-2 infection mouse model, both proteins lowered the lung viral load by up to ~1,000-fold, prevented the emergence of clinical signs in >75% animals, and increased the animal survival rate from 0% (untreated) to >87.5% (treated). These results demonstrate that both proteins are good drug candidates for protecting animals from severe COVID-19. In a head-to-head comparison of these two proteins with five previously described ACE2-Ig constructs, we found that two constructs, each carrying five surface mutations in the ACE2 region, had partial loss of neutralization potency against three SARS-CoV-2 variants. These data suggest that extensively mutating ACE2 residues near the receptor binding domain (RBD)-binding interface should be avoided or performed with extra caution. Furthermore, we found that both ACE2-Ig-95 and ACE2-Ig-105/106 could be produced to the level of grams per liter, demonstrating the developability of them as biologic drug candidates. Stress condition stability testing of them further suggests that more studies are required in the future to improve the stability of these proteins. These studies provide useful insight into critical factors for engineering and preclinical development of ACE2 decoys as broadly effective therapeutics against diverse ACE2-utilizing coronaviruses. IMPORTANCE Engineering soluble ACE2 proteins that function as a receptor decoy to block SARS-CoV-2 infection is a very attractive approach to creating broadly effective and hard-to-escape anti-SARS-CoV-2 agents. This article describes development of two antibody-like soluble ACE2 proteins that broadly block diverse SARS-CoV-2 variants, including Omicron. In a stringent COVID-19 mouse model, both proteins successfully protected >87.5% animals from lethal SARS-CoV-2 infection. In addition, a head-to-head comparison of the two constructs developed in this study with five previously described ACE2 decoy constructs was performed here. Two previously described constructs with relatively more ACE2 surface mutations were found with less robust neutralization activities against diverse SARS-CoV-2 variants. Furthermore, the developability of the two proteins as biologic drug candidates was also assessed here. This study provides two broad anti-SARS-CoV-2 drug candidates and useful insight into critical factors for engineering and preclinical development of ACE2 decoys as broadly effective therapeutics against diverse ACE2-utilizing coronaviruses.

Functional semi-finished fish product evaluation: organoleptic and evidence in vivo

Introduction The development of functional products is a new promising trend in the modern food industry. The research aims to confirm the quality indicators, efficacy and safety for living organisms of the developed functional semi-finished fish product—raw smoked sausage. The research was aimed at studying the characteristics of the semi-finished product obtained, including its organoleptic properties. However, it was also important to test the properties of the probiotic component added to the developed product and its effects in vivo on mice to verify the efficacy and safety of the E. coli 64 G strain. Methods Organoleptic and histological assessments of the product were performed. A series of experiments were also conducted to assess the product’s safety and functional properties. For this purpose, Enteracol was fed to white outbred laboratory mice with different concentrations of the active ingredient ( Escherichia coli 64G strain). Organoleptic properties of the proposed product, such as smell, consumer view, and balanced taste, demonstrated high consumer qualities of the crude smoked sausage with a probiotic component. Results The results of the controlled prospective study confirmed that the product is safe and non-toxic to living organisms: mice state alive after consuming a created meal. At the same time, assessing the product’s antagonistic activity revealed its high protective effect (85%–100% survival rate of animals in the experimental groups compared to the control with 100% mortality). Discussion The proposed product has a stimulating effect on an organism and demonstrates the antagonistic activity against pathogenic and opportunistic bacteria established in the Enterobacteriaceae family members. The next step will be a long-term study of the product’s stimulating effect to prove its positive impact on the body.

Does vitamin D reduce the mortality rate of Plasmodium infection?: a systematic review and meta-analysis

BackgroundVitamin D supplementation is recommended as an effective adjunct to counteract malaria pathogenesis, but the evidence on this point is limited and controversial. This systematic review and meta-analysis aimed to investigate the effect of vitamin D administration on the survival rate of Plasmodium-infected animals in experimentally-induced malaria on days 6 and 10 post-infection.MethodsFive electronic databases were searched up to 20 December 2021. The pooled risks ratio (RR) and associated 95% confidence interval were estimated using the Restricted-maximum likelihood (REML) random-effects model. Heterogeneity was assessed by Cochran’s Q test and I2 value. Sub-group analyses were used to identify the sources of heterogeneity for several variables, such as type of vitamin D, type of intervention, and dose of vitamin D.ResultsOut of 248 articles found in the electronic database, six were eligible for inclusion in the meta-analysis. The current study found that the pooled random effect of risks ratio favored a statistically significant effect of vitamin D administration on survival rate in infected mice on day 6 post Plasmodium infection (RR = 1.08, 95%CI 1.03, 1.15, p < 0.99; I2 = 0%). It also found that vitamin D administration significantly affected the survival rate on day 10 post-infection (RR = 1.94, 95%CI 1.39, 2.71, p < 0.001; I2 = 69.02%). Subgroup analyses demonstrated a significant pooled RRs of the positive effect of vitamin D administration for cholecalciferol (RR = 3.11, 95%CI 2.41, 4.03, p < 0.001; I2 = 0%), doses higher than 50 µg/kg (RR = 3.37, 95%CI 2.55, 4.27, p < 0.001; I2 = 0%), and oral administration (RR = 3.01, 95%CI 2.37, 3.82, p < 0.001; I2 = 0%).ConclusionThis systematic review and meta-analysis showed that vitamin D administration positively affects the survival rate in Plasmodium-infected mice. Since, the mouse model may not accurately reproduce the clinical and pathological features of human malaria, future research should investigate the impact of vitamin D in human malaria.

UP360, a Standardized Composition from Extracts of Aloe barbadense, Poria cocos, and Rosemary officinalis Protected Against Sepsis and Mitigated Acute Lung Injury in Murine Models.

Sepsis is a life-threatening organ dysfunction caused by a dysregulated and unbalanced immune response to microbial infection. Restoring immune homeostasis and infection control are considered the primary strategies to manage sepsis. Natural bioactives such as polysaccharide and polyphenols from botanicals are known for their immune modulation activity. In this study, we evaluated a standardized aloe-based composition, UP360 (constitute of polysaccharides from Aloe barbadense and Poria cocos and polyphenols from Rosemary officinalis) in lipopolysaccharide (LPS)-induced sepsis and acute inflammatory lung injury murine models. Prophylactic oral administration of UP360 for 7 days at an oral dose of 500 mg/kg improved the survival rate of mice by 62.5%, whereas all mice in the vehicle control group were deceased 82 h after LPS injection. The merit of combining these traditional herbs to yield the standardized composition UP360 was also demonstrated in this model with a mortality rate of only 30.8%, whereas 76.9%, 53.9%, and 61.5% were recorded for each individual constituents A. barbadense, P. cocos, and R. officinalis, respectively. Dose-correlated statistically significant reductions in proinflammatory cytokines and chemokine tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, and cytokine-induced neutrophil chemoattractant (CINC)-3 were observed for UP360 when administered at 250 and 500 mg/kg orally for 7 days before induction of acute lung injury (ALI) model in rats. The histopathology data from lung showed statistically significant 37.9% and 37% reductions in the overall lung damage severity and pulmonary edema, respectively, for UP360-treated rats. The aloe-based composition UP360 effectively improved the survival rate of septic animals and mitigated the severity of LPS-induced ALI in vivo. These data warrant further investigation of the composition for a potential application in human as an adjunct supplement in respiratory distress and sepsis.

Therapeutic Efficacy and Migration of Mesenchymal Stem Cells after Intracerebral Transplantation in Rats with Experimental Ischemic Stroke.

We studied therapeutic efficacy and migration characteristics of mesenchymal stem cells isolated from the human placenta after their intracerebral (stereotactic) administration to rats with the experimental ischemic stroke. It was shown that cell therapy significantly improved animal survival rate and reduced the severity of neurological deficit. New data on the migration pathways of transplanted cells in the brain were obtained.

Evaluation of the antimicrobial activity of pyrimidine compound 3-(2-benzyloxy-2-oxoethyl)quinazoline-4(3H)-oh in relation to Klebsiella pneumoniae

The study is devoted to the evaluation of the antimicrobial activity against Klebsiella pneumoniae of the pyrimidine compound 3-(2-Benzyloxy-2-oxoethyl)quinazoline-4(3H)-one under in vivo conditions in a model of generalized infection. The experiment was performed on 40 CBA line mice, which were divided into four groups: Group 1, control animals that received intraperitoneal injection water in an equivalent volume (control I); Group 2, infected animals that received no treatment (control II); Group 3, mice with generalized infection that received ceftriaxone at a dose of 50 mg/kg intraperitoneally for 7 days as treatment; Group 4, infected animals that received the study compound at a dose of 31 mg/kg (1/10 of the molecular weight) for 7 days. Generalized infection was modeled by intraperitoneal injection of Cl. pneumoniae at a dose of 3×106 in a volume of 0.5 ml. In the course of the experiment, animal survival rate was evaluated. After the mice were removed from the experiment, the blood, liver, spleen and lungs were calculated, and the total number of leukocytes, C-reactive protein and procalcitonin were determined. The compound under study was found to increase the survival rate of laboratory animals under conditions of generalized Klebsiella infection, as well as to decrease the insemination index, the total number of leukocytes and the level of markers of generalized infection. Thus, the pyrimidine derivative 3-(2-Benzyloxy-2-oxoethyl)quinazolin-4(3H)-one exhibits antibacterial activity comparable to that of the reference drug — ceftriaxone against Klebsiella pneumoniae under experimental infection.

An automatic recognition method of nematode survival rate based on bright field and dark field experimental images.

The survival rate of experimental animals is a very important index in chemical toxicity evaluation experiments. The calculation of nematode survival rate is used in many experiments. Traditional survival rate quantification methods require manual counting. This is a time-consuming and laborious work when using 384-well plate for high-throughput chemical toxicity assessment experiments. At present, there is a great need for an automatic method to identify the survival rate of nematodes in the experiment of chemical toxicity evaluation. We designed an automatic nematode survival rate recognition method by combining the bright field experimental image of nematodes and the dark field image of nematodes which is captured after adding Propidium Iodide dye, and used it to calculate the nematode survival rate in different chemical environments. Experiment results show that the survival rate obtained by our automatic counting method is very similar to the survival rate obtained by manual counting. Through several different chemical experiments, we can see that chemicals with different toxicity have different effects on the survival rate of nematodes. And the survival rate of nematodes under different chemical concentrations has an obvious gradient trend from high concentration to low concentration. In addition, our method can quantify the motility of nematodes. There are also significant differences in the motility of nematodes cultured in different chemical environments. Moreover, the nematode motility under different chemical concentrations showed an obvious gradient change trend from high concentration to low concentration. Our study provides an accurate and efficient nematode survival rate recognition method for chemical toxicology research.

Recycled polyvinyl chloride microplastics: investigation of environmentally relevant concentrations on toxicity in adult zebrafish

ABSTRACT Recycled polyvinyl chloride (PVC) microplastics have been detected in the aquatic environment. These recycled microparticles contain chemicals that are released into the environment reaching different organisms. Although the problem of the presence of recycled PVC microparticles in the environment is evident, the toxicological consequences of this contaminant to exposed organisms remains to be better determined. The aim of this study was to investigate the toxicity attributed to exposure to environmentally relevant concentrations of recycled PVC microplastics in adult zebrafish (Danio rerio). The experimental groups were: negative control, vehicle control, positive control, and recycled microplastics (20 ± 5 μm) at 5, 10 or 20 μg/L. Zebrafish (D. rerio) were exposed to respective treatments for 96 hr. Locomotion and oxidative status parameters were measured and mortality recorded. The positive control group presented increased mortality rates and decreased locomotor activity. Animals from the vehicle group did not show marked differences. Finally, no significant disturbances were found in survival rate, locomotion pattern and oxidative status of animals exposed to recycled PVC microparticles at 5, 10 or 20 μg/L. Taken together our results suggest that recycled PVC microplastics in this particle size range do not appear to exert harmful effects on exposed adult D. rerio. However, these results need to be carefully observed due to limitations including size of particle and duration of exposure parameters that might affect ecological consequences. It is suggested that additional studies applying other particles sizes and chronic exposure are needed to more comprehensively verify the toxicity of the contaminant investigated here.

Effect of mesenchymal stem cell transplantation on the structural and functional parameters of the liver in acute toxic hepatitis in rats

Acute toxic hepatitis is a serious health problem for the working population. The currently widely used pharmacological preparations with claimed hepatoprotective properties for the treatment of liver diseases have dubious therapeutic efficacy and cannot restore functional and structural changes in the damaged organ. Thus, the search for new methods of treating liver diseases, including toxic etiology, is an urgent task. An experimental study was conducted on 134 nonlinear white outbred rats (35 months) weighing 250390 g. The animals were divided into four groups: group I, control group (n = 15); group II, single intragastric administration of an oil solution of carbon tetrachloride at a dose of 1500 mg/kg (n = 41); group III, single intravenous administration of multipotent mesenchymal stem cells of the mouse (n = 20); group IV, single intragastric administration of an oil solution of carbon tetrachloride at a dose of 1500 mg/kg and subsequent intravenous transplantation of multipotent mesenchymal stem cells of the mouse (2 106) (n = 58). On days 1, 3, 5, and 7 of the experiment, blood sampling was performed for biochemical analysis, and organs and tissues were selected for histological and morphometric studies. The transplantation of multipotent mouse mesenchymal stem cells reduced the mortality of animals after exposure to carbon tetrachloride and did not cause acute toxicity. According to a morphometric study, starting from day 3, the introduction of multipotent mouse mesenchymal stem cells led to a statistically significant decrease in the infiltrative processes in the liver tissue. Statistically significant changes in the biochemical composition of the rat blood serum were noted against the transplantation of multipotent mouse mesenchymal stem cells: a decrease in the activities of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, gamma-glutamyltransferase, amylase, creatinine, uric acid, direct and total bilirubin, and cholesterol. The transplantation of multipotent mouse mesenchymal stem cells was found to be an effective method of treating acute toxic hepatitis. The use of multipotent mouse mesenchymal stem cells contributed to an increase in the survival rate of laboratory animals and faster recovery of structural and functional disorders in liver tissue after its acute toxic damage.

Study of radiation-induced pneumonitis after arc and static irradiation in lungs of rats

The clinical application of the results of the experimental radiotherapy on animals is currently not widely used. The research of radiation injuries after irradiation target in rat lungs using the image-guided platform SmART+ (Precision, North Branford, Conn., USA) is a pilot study for the establishment of an animal model of radiation-induced pulmonary toxicity.Aim: To define the typical doses and target volumes in rat lung to o serve radiation pneumonitis.Material and methods. Four groups of Wistar and Wag rats, distinguishing by age (10–16,5 months) and weight (350-850 g.), were studied. Dosimetric treatment plans were calculated prior to irradiation, using arc or static methods, doses (10–30 Gy), target lung volumes (0,045–1,24 cm3), localization (right or left, top or bottom). Dosimetric planning provides optimal radiation exposure to the target and controls organ at risk doses. Computer tomography (CT) scans were performed for 16 weeks to observe radiation pneumonitisResults. To obtain the rat lung density in Hounsfield units (HU), healthy animals were scanned, CT images were processed, and the mean and standard deviation were calculated. Lung density values of rats (control values) are 519.6 ± 46.2 (HU). Comparison of experimental data on irradiated animals and control values was carried out. As a result, the relationship of the target volume, the delivered dose and the survival rate of animals after irradiation was established. It was shown that the “mean value of lung density” could be used for quantitative analysis of lung injuries after irradiation.Conclusion. Typical doses and target volumes in rat lungs to observe radiation pneumonitis were obtained. Decreasing aeration of lung tissue was derived at delivered dose 16 Gy and irradiated target volume in lung at least 0.5 cm3 (8%).

Evaluation of the antimicrobial activity of a pyrimidine compound 2-methyl-3-(2-phenyl-2-oxoethyl) quinazolin-4(3H)-on against &lt;i&gt;Klebsiella pneumoniae&lt;/i&gt; under &lt;i&gt;in vivo&lt;/i&gt; conditions

The greatest danger is the development of multiple antimicrobial resistance in microorganisms, including Klebsiella pneumoniae, which actualizes the necessity of development and synthesis of new antimicrobial compounds. Considering the versatile pharmacological activity, pyrimidine compounds became a subject of interest for scientists in the aspect of their use as a basis for new antimicrobial agents.Aim: To assess the antimicrobial activity of the pyrimidine compound 2-Methyl-3-(2-phenyl-2-oxoethyl)quinazoline-4(3H)-on against Kl. pneumoniae under in vivo conditions.Material and Methods. The antimicrobial activity in vivo of the compound 2-Methyl-3-(2-phenyl-2-oxoethyl)quinazolin-4(3H)-on against Kl. pneumoniae was performed simulating generalized infection by intraperitoneal injection of the pathogen at a dose of 3 × 106. The experiment was conducted on CBA line mice (40 animals) divided into groups: control I – animals received intraperitoneal injection water; control II – infected animals received no treatment; experimental groups – mice with generalized infection treated with ceftriaxone at the dose 50 mg/kg intraperitoneally for 7 days, and the animals receiving the test compound at the dose 27 mg/kg against infection in the same mode. Antimicrobial activity was assessed by the following parameters: animal survival rate; internal organ and blood infestation index; total leukocyte count, C-reactive protein and procalcitonin.Results. It was found that pyrimidine derivative 2-Methyl-3-(2-phenyl-2-oxoethyl)quinazoline-4(3H)-on has a marked antimicrobial activity against Kl. pneumonia appeared in increase of animal survival in the conditions of generalized Klebsiella infection as well as in decrease of total leukocyte count, C-reactive protein and procalcitonin levels that confirms the reduction of the inflammatory reaction.Conclusion. Thus, the pyrimidine derivative 2-Methyl-3-(2-phenyl-2-oxoethyl)quinazoline-4(3H)-on exhibits antimicrobial activity, comparable with ceftriaxone, against Klebsiella pneumoniae in experimental generalized Klebsiella infection.

Therapeutic gene silencing of CKAP5 leads to lethality in genetically unstable cancer cells.

The potential of microtubule-associated protein targets for cancer therapeutics remains largely unexplored due to the lack of target-specific agents. Here, we explored the therapeutic potential of targeting cytoskeleton-associated protein 5 (CKAP5), an important microtubule-associated protein, with CKAP5-targeting siRNAs encapsulated in lipid nanoparticles (LNPs). Our screening of 20 solid cancer cell lines demonstrated selective vulnerability of genetically unstable cancer cell lines in response to CKAP5 silencing. We identified a highly responsive chemo-resistant ovarian cancer cell line, in which CKAP5 silencing led to significant loss in EB1 dynamics during mitosis. Last, we demonstrated the therapeutic potential in an in vivo ovarian cancer model, showing 80% survival rate of siCKAP5 LNPs-treated animals. Together, our results highlight the importance of CKAP5 as a therapeutic target for genetically unstable ovarian cancer and warrants further investigation into its mechanistic aspects.

A novel time-saving multiplex PCR assay for detecting and discriminating the most common canine Babesia species in Europe

In Europe, most cases of canine babesiosis are caused by Babesia canis, Babesia vogeli (large piroplasms) and Babesia vulpes (small piroplasm). Molecular diagnosis is recommended due to its high sensitivity. Species identification after sequencing allows applying a rapid and efficient treatment, leading to a better prognosis; however, it is expensive and time-consuming. Thus, the objective of the present study was to develop a time-saving multiplex polymerase chain reaction (PCR) for simultaneously detecting and discriminating between large and small forms without sequence analysis. A new multiplex PCR was designed and tested using blood samples from 79 dogs showing clinical signs compatible with babesiosis which were previously analysed using blood smears and molecular methods. Multiplex PCR successfully discriminated between both Babesia groups showing bands of 700 and 890 bp for B. canis/B. vogeli and B. vulpes, respectively. No significant differences in the results of both PCR were detected and a substantial agreement between protocols (κ = 0.64) was found. Our multiplex PCR represents a reliable tool for detecting infections by the major Babesia spp. in dogs from Europe. Since no sequence analysis is required for identifying the species involved, this PCR allows the rapid administration of an appropriate treatment, thus improving the survival rate of the infected animals. In addition, it will represent a helpful tool for unravelling the real prevalence and distribution of B. vulpes and its implication in clinical cases.

Repurposing of sevelamer as a novel antidote against aluminum phosphide poisoning: An in vivo evaluation

Aluminum phosphide (AlP) is widely used for protecting grains from pests. AlP releases toxic phosphine gas (PH3) while exposed to humidity. Poisoning with these tablets is dangerous and can cause death or serious injuries. Up to now, no definite antidote has been introduced for specific treatment of this poisoning. Sevelamer carbonate or sevelamer hydrochloride (Renagel) is a polymeric pharmaceutical prescribed for treating hyperphosphatemia in patients with chronic kidney disease. Sevelamer can bind with phosphate groups and act as an anion exchanger. Herein, sevelamer is repurposed as a potent antidote agent in phosphine gas poisoning. In vivo evaluation was conducted on male Sprague Dawley rats. The evaluation was conducted on three groups of animals: control, AlP-poisoned, and AlP-poisoned treated with sevelamer. Survival percentage, serum biomarkers level of organ injury, and ATP level were recorded. The results indicate a high survival rate in sevelamer-treated animals compared with the AlP-poisoned group (75% vs. 0% respectively, 48 h after poisoning). The analysis of serum markers of organ injury also showed that sevelamer could reduce toxicity and organ injury in poisoned animals. ATP level of separate organs showed that sevelamer treated groups were recovered. The results showed that sevelamer could be a potent antidote for managing aluminum phosphide poisoning. Moreover, a mechanism is suggested for the interaction of sevelamer with phosphine gas.

Active-targeting long-acting protein-glycopolymer conjugates for selective cancer therapy.

Non-fouling polymers are effective in improving the pharmacokinetics of therapeutic proteins, but short of biological functions for tumor targeting. In contrast, glycopolymers are biologically active, but usually have poor pharmacokinetics. To address this dilemma, herein we report in situ growth of glucose- and oligo(ethylene glycol)-containing copolymers at the C-terminal site of interferon alpha, an antitumor and antivirus biological drug, to generate C-terminal interferon alpha-glycopolymer conjugates with tunable glucose contents. The in vitro activity and in vivo circulatory half-life of these conjugates were found to decrease with the increase of glucose content, which can be ascribed to complement activation by the glycopolymers. Additionally, the cancer cell endocytosis of the conjugates was observed to maximize at a critical glucose content due to the tradeoff between complement activation and glucose transporter recognition by the glycopolymers. As a result, in mice bearing ovarian cancers with overexpressed glucose transporter 1, the conjugates with optimized glucose contents were identified to possess improved cancer-targeting ability, enhanced anticancer immunity and efficacy, and increased animal survival rate. These findings provided a promising strategy for screening protein-glycopolymer conjugates with optimized glucose contents for selective cancer therapy.

Model of small-focal ischemic cerebral infarction as a basis for the development of new methods of stroke therapy

Background. Among the existing experimental models of cerebral stroke in large animals, there is no possibility to create a focal brain infarction of limited volume, which would provide an opportunity to evaluate both the endogenous potential of neuroregeneration and to establish the effectiveness of new drugs on postischemic brain remodeling.&#x0D; Aim. Development of a method for modeling small-focal ischemic infarction of the cerebral cortex in a mini-pig.&#x0D; Material and methods. The study used mature female mini-pigs of the Vietnamese fold-bellied breed. Two-level occlusion of the great vessels was performed by ligation of the right common carotid artery and subsequent cauterization of the distal branches of the middle cerebral artery. In the postoperative period, the survival rate of animals and neurological deficit were assessed. On the 7th day, the animals were taken out of the experiment, the brain was isolated from the cranial box. Macroscopic and microscopic examinations included analysis of the localization and area of cerebral infarction, as well as morphological changes in the infarcted and peri-infarcted areas of the cerebral cortex.&#x0D; Results. With a 100% survival rate of experimental animals on the 3rd day after the operation, a neurological examination revealed violations of skin sensitivity and muscle tone in the hind and fore limbs, which partially recovered a week after the onset of ischemic stroke. Macroscopic examination of the brain on the 7th day after the operation visually revealed a small-focal ischemic cerebral infarction in the parietal lobe of the left hemisphere. Histological analysis of the cerebral cortex revealed a wedge-shaped necrotic focus of ischemic injury. In the peri-infarction region of the brain, reactive tissue changes with preserved nerve cells, mostly without visible damage, were found.&#x0D; Conclusion. The developed two-level method of stroke modeling in a mini pig induces ischemic cerebral infarction of limited volume in the parietal lobe; non-critical histological changes in the peri-infarct area and a slight neurological deficit suggest the possibility of using this model to develop new methods of stroke therapy.

A MODIFIED SURGICAL SEPSIS MODEL SATISFYING SEPSIS-3 AND HAVING HIGH CONSISTENCY OF MORTALITY.

Background : Cecal ligation and perforation (CLP) is currently considered the criterion standard model of sepsis; however, there are some deficiencies, such as low clinical relevance, inconsistency in severity grading, and an unknown proportion of CLP animals meeting the requirements of sepsis-3. Methods : Adult rats were randomly divided into the following three groups: modified CLP (M-CLP) group, CLP group, and sham group. The vital organ function of rats was evaluated 24 hours postoperatively by blood pressure, behavioral testing, histopathology, and blood test. Cytokine levels were determined by enzyme-linked immunosorbent assay, and T-cell suppression was assessed by flow cytometry. The stability of the model was evaluated by comparing the survival rates of repeated experiments in all groups from day 1 to day 14. Results : More rats in the M-CLP group met Sepsis-3 criteria than those in the CLP group 24 hours postoperatively (53.1% vs. 21.9%, P = 0.01). Rats in the M-CLP group developed more serious hepatic, pulmonary, and renal dysfunction. Similar to human sepsis, rats in the M-CLP group demonstrated more serious immunosuppression and systemic inflammation compared with the CLP group. In addition, disease development and severity, which was indicated by the stable survival rates of model animals, were more stable in the M-CLP group. Conclusions : More rats could meet Sepsis-3 criteria with this novel surgical procedure, which may reduce the number of animals needed in preclinical sepsis experiments. This stable M-CLP model may contribute to the development of new therapies.