Life Cycle and Transmission of Cyclospora cayetanensis: Knowns and Unknowns

Although infections with Cyclospora cayetanensis are prevalent worldwide, many aspects of this parasite’s life cycle and transmission remain unknown. Humans are the only known hosts of this parasite. Existing information on its endogenous development has been derived from histological examination of only a few biopsy specimens. Its asexual and sexual stages occur in biliary-intestinal epithelium. In histological sections, its stages are less than 10 μm, making definitive identification difficult. Asexual (schizonts) and sexual (gamonts) are located in epithelial cells. Male microgamonts have two flagella; female macrogametes contain wall-forming bodies. Oocysts are excreted in feces unsporulated. Sporulation occurs in the environment, but there are many unanswered questions concerning dissemination and survival of C. cayetanensis oocysts. Biologically and phylogenetically, C. cayetanensis closely resembles Eimeria spp. that parastize chickens; among them, E. acervulina most closely resembles C. cayetanensis in size. Here, we review known and unknown aspects of its life cycle and transmission and discuss the appropriateness of surrogates best capable of hastening progress in understanding its biology and developing mitigating strategies.

Efficacy of Vanadyl Sulfate and Selenium Tetrachloride as Anti-Diabetic Agents against Hyperglycemia and Oxidative Stress Induced by Diabetes Mellitus in Male Rats

The use of metals in medicine has grown in popularity in clinical and commercial settings. In this study, the immune-protecting effects and the hypoglycemic and antioxidant activity of vanadyl sulfate (VOSO4) and/or selenium tetrachloride (Se) on oxidative injury, DNA damage, insulin resistance, and hyperglycemia were assessed. Fifty male albino rats were divided into five groups, and all treatments were administrated at 9:00 a.m. daily for 60 successive days: control, STZ (Streptozotocin; 50 mg/kg of STZ was given to 6 h fasted animals in a single dose, followed by confirmation of diabetic state occurrence after 72 h by blood glucose estimation at >280 mg/dl), STZ (Diabetic) plus administration of VOSO4 (15 mg/kg) for 60 days, STZ (Diabetic) plus administration of selenium tetrachloride (0.87 mg/Kg), and STZ plus VOSO4 and, after 1/2 h, administration of selenium tetrachloride at the above doses. The test subjects’ blood glucose, insulin hormone, HbA1C, C-peptide, antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, myeloperoxidase, and xanthine oxidase), markers of lipid peroxidation (MDA), and histological sections of pancreatic tissues were evaluated, and a comet assay was performed. Histological sections in pancreas tissues were treated as indicators of both VOSO4 and selenium tetrachloride efficacy, either alone or combined, for the alleviation of STZ toxicity. The genotoxicity of diabetes mellitus was assessed, and the possible therapeutic roles of VOSO4 or selenium tetrachloride, or both, on antioxidant enzymes were studied. The findings show that the administration of VOSO4 with selenium tetrachloride reduced oxidative stress to normal levels, lowered blood glucose levels, and elevated insulin hormone. Additionally, VOSO4 with selenium tetrachloride had a synergistic effect and significantly decreased pancreatic genotoxicity. The data clearly show that both VOSO4 and selenium tetrachloride inhibit pancreatic and DNA injury and improve the oxidative state in male rats, suggesting that the use of VOSO4 with selenium tetrachloride is a promising synergistic potential ameliorative agent in the diabetic animal model.

Immunolocalization of FOXP3, JAK1 and STAT5 in Preeclamptic, Intrauterine Growth Restricted and Gestational Diabetic Human Placentas

Objective: The aim of the study to show the relation of T cells in placental villous fragments with FOXP3,JAK1 and STAT5 receptors in different conditions such as GDM, PE and IUGR placental tissues. Methods: Specimens of ten(10) diabetic placentas, ten(10) preeclamptic, ten(10) intrauterine growth restricted placentas and ten(10) control placentas were collected by systematic uniform random sampling. Immunohistochemical detections of FOXP3, JAK1 and STAT5 were performed in histological sections for each group’s placental tissue. The H-score value was derived for each specimen by calculating the sum of the percentage of syncytiotrophoblast and syncytial nodes in placenta and intervillus area. They were categorized by intensity of staining, multiplied by its respective score. Results: FOXP3, JAK1 and STAT5 immunoreactivity comparisons are shown in four groups of placentas. FOXP3 immunoreactions significantly increase in GDM group. JAK1 and STAT5 immunoreactions significantly decrease in PE group. STAT5 immunoreactivity was detected crucially increase in GDM group. Discussion: The results showed that in different conditions such as PE,GDM and IUGR, T cells in placental villous fragments have relation with FOXP3,JAK1 and STAT5 receptors and that FOXP3 can inactivate the PE and IUGR in the placental tissue. We have also confirmed as other studies that JAK-STAT pathway plays important role in PE,IUGR and GDM placental tissue.

Forensic determination of determination of damage to human internal organs by the method of reconstruction of the optical activity of histological sections

The article presents the results of experimental testing of the technique of tomographic reconstruction of circular birefringence maps of molecular complexes in a digital histological study of the age of damage to tissues of internal organs (brain, liver and kidney) of temporary monitoring of the mean value, dispersion, asymmetry and kurtosis, characterizing the distribution of the magnitude of the optical activity of histological sections of the brain, liver and kidney identification of the temporal extent of linear changes in statistical parameters and the accuracy of determining the duration of damage to human internal organs by digital histological methods of polarization reconstruction (tomography) of circular birefringence of molecular complexes. Aim of the work. Development of a technique for determining the duration of damage to human internal organs by digital histological methods of polarization reconstruction of circular birefringence of molecular complexes. Materials and methods. The object of the study was the histology of samples of human internal organs (brain, kidney and liver) with different duration of damage from 1 hour to 120 hours. For control, we used BT samples of those who died from coronary artery disease with different duration of damage from 1 hour to 120 hours. The studies were carried out using the technique of polarization reconstruction of circular birefringence of molecular complexes. Results. A set of treatment-relevant relationships between temporal changes in the statistical structure of topographic maps of circular birefringence of optically active molecular complexes of histological sections of human internal organs with different duration of damage and variations in the mean value, dispersion, asymmetry and kurtosis, characterizing the distribution of the value of this parameter of anisotropy, has been determined. Conclusions. A new original method has been developed for tomography of the optical activity of molecular complexes of tissues of human internal organs in a digital histological study of the age of damage to the tissues of the brain, liver and kidney, as well as the myocardium and lung tissue at a time interval of 1 hour. up to 120 hours

FORENSIC MEDICAL CRITERIA OF DIFFERENTIAL DIAGNOSIS OF ALCOHOL AND CARBON MONOXIDE POISONING BY USING AZIMUTHAL-INVARIANT POLARIZATION MUELLER-MATRIX MICROSCOPY

Introduction: Fatal intoxications with alcohol and carbon monoxide are the most common causes of poisoning that occur in forensic medical practice. Therefore, the differential diagnosis of these intoxications is extremely important. To solve this problem, we propose the use of the method of azimuthal-invariant polarization Mueller-matrix microscopy of histological sections of biological tissues (BT). Objectives: The purpose of the study was to establish statistically significant criteria for differentiation of the cause of death in case of alcohol poisoning (AP) and carbon monoxide poisoning (CMP) by using azimuthal-invariant polarization Mueller-matrix microscopy of polycrystalline fractions of histological sections of organs and blood films of the human body. Results: A number of characteristic features have been identified that have not been previously described and allow to objectively differentiate the cause of death in the case of AP and CMP, namely Ek – 0.95±0.045 and 1.81±0.092 in the study of histological sections of the myocardium; Ek – 0,19±0,011 and 1,11±0,094 – polycrystalline blood films. Analysis of the obtained data revealed a good level of accuracy in the study of histological sections of the myocardium (As 85%) and polycrystalline blood films (As 88%). Conclusion: Thus, developed and tested method of azimuthal-invariant polarization Mueller-matrix microscopy allows to differentiate the cause of death in cases of AP and CMP.

Differential diagnostics of the presence of damage by the method of digital polarization mapping of microsocopy images of hystological sections of human organs

The article presents the results of experimental testing of methods of azimuthal-invariant polarization mapping of microscopic images of samples of histological sections of the myocardium and lung tissue; time monitoring of changes in the magnitude of statistical moments of the 1-4th orders, characterizing the distributions of the azimuth and ellipticity of polarization of microscopic images of myocardial and lung tissue samples with different age of damage; determination of the diagnostic efficiency (time interval and accuracy) of establishing the age of damage to human internal organs by digital histological methods of mapping azimuth maps and polarization ellipticity of microscopic images. Aim of the work. Development of a digital histology technique for samples of human internal organs. Materials and methods. The object of the study was the histology of samples of human internal organs (myocardium, lung tissue) with different duration of damage from 1 hour to 120 hours. For control, we used BT samples from those who died from coronary artery disease with different duration of damage from 1 hour to 120 hours. The studies were carried out using the method of digital polarization mapping of microscopic images of histological sections of biological tissues of human internal organs. Results. Revealed the following ranges ramp variation quantity statistics polarizing digital histology and accuracy of the limitation of damage: a. Arts azimuth polarization microscopic image of a magnification ×40 (asymmetry – 12 hours, kurtosis – 12 hours, accuracy – 55-60 min), maps of ellipticity of polarization of microscopic images with a magnification of ×40 (asymmetry – 12 hours, kurtosis – 12 hours, accuracy – 65-75 min). Conclusions. Scenarios of changes in the statistical structure of maps of azimuth and ellipticity of polarization of microscopic images of histological sections of human internal organs have been determined - with an increase in the duration of damage, the value of the mean and variance decreases, the asymmetry and kurtosis increase.

Time since hemorrhages of traumatic and non-traumatic genesis into the substance of the human brain formation using distribution of linear dichroism fluctuations reconstruction

For a forensic expert-practitioner, it is especially important to objectively diagnose and time since the formation of hemorrhage (TSFH) in the substance of the human brain (SHB) of traumatic and non-traumatic origin, as there are cases when the external examination of the corpse at the scene are absent, and at internal research find hemorrhages in a brain. In forensic practice, to verify the cause of death, physical-optical methods are successfully used, which are based on laser irradiation of biological tissues with subsequent mathematical and statistical processing of the obtained data. Previous studies on the possibility of differentiating the cause of death by traditional polarization methods have yielded positive results, which suggests the possibility of their suitability for verification of the genesis of hemorrhage into the brain. For a forensic expert-practitioner, the main thing is objectivity, accuracy and speed of obtaining the result, which could fully satisfy the methods of laser polarimetry in the case of determining the TSFH of traumatic and non-traumatic origin in SHB. Therefore, it is necessary to continue the development and research of these methods for this purpose. Aim of the work. To substantiate the possibility of using the method of differential Mueller-matrix mapping of phase anisotropy to determine the temporal dynamics of maps of linear birefringence of histological sections of human brain in determining the age of hemorrhage in human brain substance and to develop forensic criteria for determining the age. death due to cerebral infarction of ischemic and hemorrhagic origin. Materials and methods. To achieve this goal, we studied native histological preparations SHB from 130 corpses with a known time of death. The cause of death was TBI (group II (n=35)), cerebral infarction of ischemic origin (group III (n=32)), hemorrhagic stroke (group IV (n=33)), acute coronary insufficiency (group I – comparison group (n=30)). The values of the distribution of the coordinates of the polarization parameters at the points of the microscopic images at the location of the standard Stokes polarimeter were measured. Experimental measurements of Stokes-parametric images of biological layers were performed according to the method presented in the sources. Subsequently, the obtained data were subjected to statistical processing and evaluation of the obtained results. Statistical moments (SM) of the 1st-4th orders (mean (SM1), variance (SM2), asymmetry (SM3) and excess (SM4)) of each map were determined. Results and discussion. Comparative analysis of polarization Mueller-matrix mapping images of SHB sections from all groups revealed the destruction of the polycrystalline structure formed by optically active protein complexes of the brain substance, which indicates a decrease in absolute values and range of their scatter with increasing hemorrhage time. This is indicated by the coordinate inhomogeneity of the Mueller-matrix invariant maps of histological sections of SHB of all groups. For histograms that characterize the distributions of the Mueller-matrix invariant samples from all (comparison groups 1 and experimental 2-4) groups, are characterized by individual and significant variations in the values of statistical moments. Due to this, with increasing hemorrhage time, the value of the mean (SM1) and variance (SM2) decreases. Asymmetry (SM3) and excess (SM4), on the contrary, increase. The analysis of the results of statistical processing of the topographic structure of LD tomograms of fibrillar networks of histological sections of SHB dead from all groups shows a greater temporal dynamics of necrotic destruction of nervous tissue. Accordingly, there is a faster time decrease in the absolute values and the range of scatter of the LD value with increasing TSFH. That is, the diagnostic sensitivity of the statistical moments of the 3rd and 4th orders for azimuthal-invariant Mueller-matrix differentiation of nerve tissue samples of the brain of the deceased of control group 1 and all experimental groups 2-4 (p<0,05) was revealed. Conclusions. A series of studies of the effectiveness of a new in forensic practice method of differential Mueller-matrix mapping of partially depolarizing histological sections of SHB and tomographic reproduction of optical anisotropy parameters of their polycrystalline structure revealed a high level of accuracy of differentiation and formation of genesis, even under conditions of small geometric thickness of experimental samples. The range of linear change of values of statistical moments of the 1st - 4th orders which characterize distributions of size of LD of fibrillar networks of histologic sections of SHB of the dead from all groups, makes 24 h. In the range of 6-24 hours, the accuracy of determining the TSFH using statistical processing of the topographic structure of LD tomograms of fibrillar networks of histological sections of TSFH is (30±5) minutes.

MENINGOENCEPHALITIS IN BROILER CHICKS DUE TO ASPERGILLUS FUMIGATUS INFECTION

Forty five cases, 7-14 day-old chicks were studied, they showed respiratory and nervous signs with yellowish granulomatous necrotic foci on the air sacs, liver, kidneys, cerebellum and cerebrum. Swabs were taken 7 days post inoculation, culture was made on Sabouraud's dextros agar,. Aspergillus fumigatus was isolated. Diagnosis was confirmed by experimentally exposed one day-old chicks to aerosols of A. fumigatus conidia and also by microscopic demonstration of typical fruiting bodies and spores of the fungus in fresh preparations made from Sabouraud's agar. tissues from fungal-infected Examination of histological sections of cerebrum and cerebellum chicks stained with hematoxylin and eosin and PAS stains revealed granulomas lesion with a central area of necrosis containing heterophils surrounded by macrophages, gaint cells, lymphocytes and layer of fibrous tissue. It was concluded that the cases deal within this study had been infected with A. fumigataus.