Effects of vitrification techniques on the formation of skin cryobank of the ocelot (Leopardus pardalis).

BACKGROUND: Skin cryobanks represent important tools for the conservation of the maximum genetic representation of a population, especially those with a certain degree of threat to extinction, such as the ocelot. A relevant step towards the proper establishment of these banks is the definition of adequate cryopreservation techniques for the conservation of the skin. OBJECTIVE: We evaluated the effects of two different techniques [direct vitrification in cryovials (DVC) and solid-surface vitrification (SSV)] for the preservation of ear skin derived from ocelot. MATERIALS & METHODS: For both techniques, we vitrified the ear skin using Dulbeccos modified Eagles medium with 3.0 M dimethyl sulfoxide, 0.25 M sucrose, and 10% fetal bovine serum. Non-cryopreserved tissues were used as control (control group). All tissues were analyzed for their morphometric characteristics by conventional histology and morphological / functional analysis by cell ability during the culture. RESULTS: While tissues cryopreserved by DVC showed similar values for dermis thickness and number of perinuclear halos to the control, tissues cryopreserved by SSV showed similarities to the control regarding the number of melanocytes, percentage of collagen fibers, and numbers of viable cells by apoptosis analysis. Additionally, none of the vitrification techniques affected stratum corneum thickness, number of keratinocytes, tissue proliferative activity, cell viability, or metabolism. CONCLUSION: Both vitrification techniques (DVC and SSV) can be used for the conservation of ocelot skin; however, SSV guarantees a higher cellular quality after in vitro tissue culture in most of the parameters evaluated, such as viability, metabolism, and apoptosis analysis. doi.org/10.54680/fr23110110412.

Evaluation of different cryoprotectant solutions for the cryopreservation of somatic tissues of Dasyprocta leporina (Linnaeus, 1758).

BACKGROUND: Somatic tissue banks represent important tools for the conservation of wild mammals, aiming at the immediate maintenance and safeguarding of biological samples. For agouti, Dasyprocta leporina, studies on the formation of these banks are still scarce, especially regarding protocols of the best cryoprotectant solution employed. OBJECTIVE: To optimize the cryoprotectant solution [ethylene glycol (EG), dimethyl sulfoxide (DMSO), sucrose (SUC)] used for the cryopreservation of agouti somatic tissues. MATERIALS AND METHODS: We treated ear tissues with various cryoprotectant solutions: 3.0 M EG (EG group), 3.0 M EG and 0.25 M SUC (EG-SUC group), 3.0 M DMSO (DMSO group), 3.0 M DMSO and 0.25 M SUC (DMSO-SUC group), 1.5 M EG and 1.5 M DMSO (EG-DMSO group) and 1.5 M EG, 1.5 M DMSO and 0.25 M SUC (EG-DMSO-SUC group). Non-cryopreserved tissues were used as controls. All tissues were analyzed for their ultrastructural and morphometric characteristics by scanning electron microscopy and conventional histology. RESULTS: EG-DMSO-SUC was found to be the optimal cryoprotectant solution in terms of the evaluated parameters, such as thickness of the dermis and skin, number of perinuclear halos, proliferative potential, number of empty lacunas and degenerated chondrocytes. CONCLUSION: Agouti somatic tissue cryopreservation may serve for its conservation and as an experimental model for the development of preservation methods for species of the same genus that are either vulnerable or critically endangered.

Twin screw granulation: A simpler re-derivation of quantifying fill level.

The fill level is defined as the volume occupied by the powder and granules inside the twin-screw granulator in proportion to the maximum barrel channel void 'free' volume. In literature, the fill level is one of the key factors that determine the final granule properties as it relies on several factors such as the screw speed, screw element geometry, mass flow rate and barrel length. However, quantitative prediction of the fill level in twin-screw granulation (TSG) is still a developing area, which is required to enable effective development of process design space, to yield a product with desired quality attributes for all process scale levels (small to large equipment). In this study, a simple geometrical model is presented that predicts the barrel channel fill level in TSG. This model relates the volumetric flow rate to the forward volumetric conveying rate of the screws when they advance in the axial direction. Experimentation was conducted to validate the model by analytically measuring mass hold-up, the amount of material remaining in the barrel after steady state was reached, as the fill level is proportional to mass hold-up. Furthermore, the trends in the extent of granulation with the proposed model were investigated. Good agreement was found between the proposed fill level model and the mass hold-up for various screw elements, therefore the model provides a more practical measure of the fill level in TSG.

Cryopreservation of Collared Peccary (Pecari tajacu Linnaeus, 1758) Somatic Cells is Improved by Sucrose and High Concentrations of Fetal Bovine Serum.

BACKGROUND: The formation of somatic cell banks is affected by, amongst other factors, the cryoprotectant solution used. The selection of an effective solution, therefore, is a primary parameter. OBJECTIVE: We optimized the cryoprotectant used for collared peccary somatic cell cryopreservation. MATERIALS AND METHODS: We categorized cells into different groups based on their cryopreservation and evaluated the morphology, viability, proliferative activity, metabolism, and oxidative stress. One group was cryopreserved in 10% DMSO with 10% fetal bovine serum (DMSO-10FBS), and another with 50% FBS (DMSO-50FBS). The cryopreservation of both groups included the presence of 0.2 M sucrose (DMSO-SUC-10FBS and DMSO-SUC-50FBS). Non-cryopreserved cells and cells cryopreserved with 10% DMSO (DMSO) supplemented with 0.2 M sucrose (DMSO-SUC) were used as controls. RESULTS: There was no difference observed in morphology or viability among the groups. Proliferative activity was reduced in DMSO-10FBS when compared to controls. Although cryopreservation reduced metabolism, no difference was observed among solutions. A lower level of reactive oxygen species was observed in cells of DMSO-SUC-50FBS when compared to other cryoprotectants. Only cells of DMSO-SUC-50FBS had mitochondrial potential similar to non-cryopreserved cells. CONCLUSION: 10% DMSO supplemented with 50% FBS and 0.2 M SUC was observed to be the most efficient cryoprotectant for preserving collared peccary somatic cells.

Wetting of binary powder mixtures.

The wetting process involved when a liquid droplet comes into contact with a mixture of particles is a complex phenomenon which is often understood by reference to Cassie-Baxter theory. However, various authors have applied the Cassie-Baxter theory for the prediction of contact angles on two-component mixtures without success. We hypothesise that the main difficulty in applying the Cassie-Baxter theory to mixtures is that if the particles differ in size, it is possible for the small particles to coat the large particles, so reducing the available surface area of the large particles. This leads to the view that bulk volume fractions are not good estimates of surface fractions of the components within the mixture. We argue that the Cassie-Baxter theory over represents the influence of large particles and that below a certain critical volume fraction they exert no influence. We present a simple geometrical model that relates the critical surface coverage volume fraction to the Sauter mean particle size of the binary mixture components. As a consequence, the wetting behaviour can be determined from the bulk volume fractions and the calculated critical surface coverage volume fraction, by means of a simple geometric model. We show that the simple model describes the five two-components systems reported here and a further four systems reported in the literature, irrespective of whether the larger or small particles are hydrophobic/hydrophilic. With this model, it is possible to predict the wetting behaviour of mixtures of particles that coat each other using very simple characterisation methods, so reducing the development time in the creation of formulations in the pharmaceutical industry.

Assessing Particle Segregation Using Near-Infrared Chemical Imaging in Twin Screw Granulation.

In the present study the application of near-infrared chemical imaging (NIR-CI) for assessing particle segregation in granules from continuous twin screw granulation (TSG) granules, were the complex attributes of the machinery configuration in relation to particle segregation is not well understood was investigated. Experiments were performed along the compartmental length of the TSG barrel channel by varying the screw element type and liquid binder viscosity. Examination of the data showed a direct correlation between dispersion due to shear force and de-mixing of particles, which allowed for identification of fundamental granule segregation mechanisms affecting content uniformity in TSG. Particle segregation behavior was linked to dispersion due to shear force through a proposed regime mapping approach which links de-mixing potential to controlling granule formation mechanisms with a new dimensionless mixing number. This was carried out in order to provide a general guideline of how particles segregate along the length of the TSG barrel channel.

Epidemiologic Profile of Patients Transplanted With Hematopoietic Stem Cells in a Reference Service in the State of Rio Grande do Norte, Brazil.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) consists of the intravenous infusion of healthy hematopoietic stem cells to restore the medullary and immunologic function of patients affected by a series of hematologic, oncologic, immunologic, malignant and nonmalignant inherited or acquired diseases, with the possibility of cure or increase of disease-free survival. OBJECTIVE: To characterize the epidemiologic profile and the cases of death of patients who underwent HSCT. METHODS: This is a cohort quantitative study, nested with a retrospective, descriptive, and analytical study of a hospital-based cohort that included the patients who underwent HSCT at a referral service in the state of Rio Grande do Norte, a region of northeastern Brazil. RESULTS: There was a slight male prevalence (52.94%), the age of the patients ranged from 2 to 73 years old, 18.38% were brown, 47.06% were married, 15.07% were students, 78.31% had a diagnosis of multiple myeloma, 93.38% developed gastrointestinal toxicities, all patients received chemotherapeutic treatment, 54.78% had allogeneic HSCT, and the cause of the most recorded deaths was septic shock (48.19%). CONCLUSIONS: This study showed relevant scientific evidence on the clinical and epidemiologic profile of patients who underwent HSCT. In general, sociodemographic data are similar to national and international research results.

Hypermanganesemia due to mutations in SLC39A14: further insights into Mn deposition in the central nervous system.

BACKGROUND: The SLC39A14, SLC30A10 and SLC39A8 are considered to be key genes involved in manganese (Mn) homeostasis in humans. Mn levels in plasma and urine are useful tools for early recognition of these disorders. We aimed to explore further biomarkers of Mn deposition in the central nervous system in two siblings presenting with acute dystonia and hypermanganesemia due to mutations in SLC39A14. These biomarkers may help clinicians to establish faster and accurate diagnosis and to monitor disease progression after chelation therapy is administered. RESULTS: A customized gene panel for movement disorders revealed a novel missense variant (c.311G > T; p.Ser104Ile) in SLC39A14 gene in two siblings presenting at the age of 10 months with acute dystonia and motor regression. Mn concentrations were analyzed using inductively coupled mass spectrometry in plasma and cerebrospinal fluid, disclosing elevated Mn levels in the index case compared to control patients. Surprisingly, Mn values were 3-fold higher in CSF than in plasma. We quantified the pallidal index, defined as the ratio between the signal intensity in the globus pallidus and the subcortical frontal white matter in axial T1-weighted MRI, and found significantly higher values in the SLC39A14 patient than in controls. These values increased over a period of 10 years, suggesting the relentless pallidal accumulation of Mn. Following genetic confirmation, a trial with the Mn chelator Na2CaEDTA led to a reduction in plasma Mn, zinc and selenium levels. However, parents reported worsening of cervical dystonia, irritability and sleep difficulties and chelation therapy was discontinued. CONCLUSIONS: Our study expands the very few descriptions of patients with SLC39A14 mutations. We report for the first time the elevation of Mn in CSF of SLC39A14 mutated patients, supporting the hypothesis that brain is an important organ of Mn deposition in SLC39A14-related disease. The pallidal index is an indirect and non-invasive method that can be used to rate disease progression on follow-up MRIs. Finally, we propose that patients with inherited defects of manganese transport should be initially treated with low doses of Na2CaEDTA followed by gradual dose escalation, together with a close monitoring of blood trace elements in order to avoid side effects.

Asymptomatic Transmission and the Dynamics of Zika Infection.

Following the 2013-14 outbreak in French Polynesia, the Zika virus (ZIKV) epidemic spread widely to many countries where Aedes Aegypti as the main transmitting vector is endemic. The lack of a second wave of ZIKV infection in most affected regions may suggest that a sufficiently high level of herd immunity was reached during the first wave. We developed an agent-based transmission model to investigate the role of asymptomatic infection on the likelihood of observing a second wave, while accounting for its relative transmissibility. We found that, as the relative transmissibility of asymptomatic infection increases, a second wave is more likely to occur, despite an increase in the attack rate during the first wave. When the reproduction number varies between 1.9 and 2.8 based on estimates for Antioquia, Colombia, the attack rate varies between 4% and 26% for a low (below 10%) effectiveness of interventions in blunting the ZIKV transmission from symptomatic cases to mosquitoes. Moreover, the fraction of cases due to sexual transmission is estimated below 4% of the cumulative incidence. Our analyses underscore the need to quantify the transmissibility of asymptomatic infections, without which the overall attack rates and the level of herd immunity cannot be accurately estimated.

Strategies for halting the rise of multidrug resistant TB epidemics: assessing the effect of early case detection and isolation.

Background: The increasing rates of multidrug resistant TB (MDR-TB) have posed the question of whether control programs under enhanced directly observed treatment, short-course (DOTS-Plus) are sufficient or implemented optimally. Despite enhanced efforts on early case detection and improved treatment regimens, direct transmission of MDR-TB remains a major hurdle for global TB control. Methods: We developed an agent-based simulation model of TB dynamics to evaluate the effect of transmission reduction measures on the incidence of MDR-TB. We implemented a 15-day isolation period following the start of treatment in active TB cases. The model was parameterized with the latest estimates derived from the published literature. Results: We found that if high rates (over 90%) of TB case identification are achieved within 4 weeks of developing active TB, then a 15-day patient isolation strategy with 50% effectiveness in interrupting disease transmission leads to 10% reduction in the incidence of MDR-TB over 10 years. If transmission is fully prevented, the rise of MDR-TB can be halted within 10 years, but the temporal reduction of MDR-TB incidence remains below 20% in this period. Conclusions: The impact of transmission reduction measures on the TB incidence depends critically on the rates and timelines of case identification. The high costs and adverse effects associated with MDR-TB treatment warrant increased efforts and investments on measures that can interrupt direct transmission through early case detection.

Analysis of risk factors and prevalence of haemoplasma infection in dogs.

Mycoplasma haemocanis (Mhc) and 'Candidatus Mycoplasma haematoparvum' (CMhp) are canine haemoplasma species that can induce anaemia in immunocompromised and/or splenectomised dogs. This study aimed to determine the prevalence and phylogeny of canine haemoplasma species in dogs from Nigeria and describe any risk factors for infection. Canine haemoplasma species-specific and generic haemoplasma qPCR assays were used. The species-specific qPCR assays found Mhc infection in 18 of 245 dogs (7.3%), and CMhp infection in only one dog (0.4%). The generic haemoplasma qPCR assays were positive in 44 of 245 (17.9%) dogs. Twenty-five dogs had discordant qPCR results in that they were generic haemoplasma qPCR positive but species-specific qPCR negative. Further evaluation of these dogs by 16S rDNA sequencing gave limited results but 5 were confirmed to be infected with non-haemoplasma species: 2 Anaplasma phagocytophilum, 1 Anaplasma ovis, 1 Serratia marcescens and 1 Aerococcus spp. The 16S rRNA gene sequences from Mhc species showed>99.8% identity with each other and>99.6% identity with GenBank sequences, and resided in a single clade with other global Mhc and Mycoplasma haemofelis sequences, indicating low 16S rRNA genetic variability amongst this canine haemoplasma species.

Real-time assessment of relapse risk based on the WT1 marker in acute leukemia and myelodysplastic syndrome patients after hematopoietic cell transplantation.

Relapse is the major cause of treatment failure after allogeneic hematopoietic cell transplantation (alloHCT) for acute leukemia and myelodysplastic syndrome (MDS). Wilms' tumor Ag (WT1) is overexpressed in the majority of acute leukemia and MDS patients and has been proposed as a universal diagnostic marker for detection of impending relapse. Comprehensive studies have shown that WT1 transcript levels have predictive value in acute leukemia patients in CR after chemotherapy. However, the focus of this study is the period after alloHCT for predicting relapse onset. We analyzed the accumulation of WT1 mRNA transcripts in PB of 82 leukemia and MDS patients and defined specific molecular ratios for relapse prediction. The extensively validated WT1/c-ABL ratio was used to normalize increases in WT1 transcript levels. The observed lead time of crossing or exceeding set WT1 levels is presented along with linear interpolation to estimate the calculated day the WT1 thresholds were crossed. The WT1/c-ABL transcript ratio of 50 or above yielded 100% specificity and 75% sensitivity reliably predicting future relapse with an observed average of 29.4 days (s.d.=19.8) and a calculated average of 63 days (s.d.=29.3) lead time before morphologic confirmation. A lower ratio of 20 or above gave lower specificity, but higher sensitivity (84.8% and 87.5%, respectively) identified more patients who relapsed, at earlier times, providing an earlier warning with actual average lead time of 49.1 days (s.d.=30.8) and calculated average of 78 days (s.d.=28.8). WT1 transcript levels serve as a diagnostic relapse test with greater sensitivity than the morphologic approach used in the clinic as a readout.

[Blue rubber bleb nevus syndrome: A case report]. / Síndrome de nevos azules ahulados: reporte de un caso.

Blue Rubber Bleb Nevus Syndrome (BRBS) is a rare disease, characterized by multiple vascular malformations in the skin and gastrointestinal tract. Other organs can also be affected, presenting different clinical manifestations such as arthralgia, epistaxis, hemoptysis, hematuria, hemothorax, mild thrombocytopenia, consumptive coagulopathy, and bone deformities, among others. We present a case of BRBS in a nine-year-old boy with the characteristic clinical manifestations of punctated purplish-blue skin lesions that vary in size and gastrointestinal vascular malformations with upper digestive tract bleeding.

Responses of antioxidant systems after exposition to rare earths and their role in chilling stress in common duckweed (Lemna minor L.): a defensive weapon or a boomerang?

Extensive agriculture application of rare earth elements (REEs) in Far East countries might cause spreading of these metals in aquatic and terrestrial ecosystems, thus inducing a growing concern about their environmental impact. In this work the effects of a mix of different REE nitrate (RE) and of lanthanum nitrate (LA) on catalase and antioxidant systems involved in the ascorbate-glutathione cycle were investigated in common duckweed Lemna minor L. The results indicated that L. minor shows an overall good tolerance to the presence of REEs in the media. Treatments at concentrations up to 5 mM RE and 5 mM LA did not cause either visible symptoms on plants or significant effects on reactive oxygen species (ROS) production, chlorophyll content, and lipid peroxidation. Toxic effects were observed after 5 days of exposition to 10 mM RE and 10 mM LA. A remarkable increase in glutathione content as well as in enzymatic antioxidants was observed before the appearance of the stress symptoms in treated plants. Duckweed plants pretreated with RE and LA were also exposed to chilling stress to verify whether antioxidants variations induced by RE and LA improve plant resistance to the chilling stress. In pretreated plants, a decrease in ascorbate and glutathione redox state and in chlorophyll content and an increase in lipid peroxidation and ROS production levels were observed. The use of antioxidant levels as a stress marker for monitoring REE toxicity in aquatic ecosystems by means of common duckweed is discussed.

Previsão Bayesiana de valores genéticos de touros por meio do modelo auto-regressivo para dados em painel / Bayesian forecasting of sires breeding values using autoregressive panel data model

Inferência Bayesiana do modelo auto-regressivo foi aplicado para dados em painel a dados reais de DEPïs de touros da raça Nelore publicadas entre 2000 e 2005. Na análise foram consideradas a função de verossimilhança exata e a obtenção de distribuições preditivas de dados futuros. Verificou-se ser recomendável o agrupamento dos animais em grupos homogêneos de acordo com a acurácia. Constatou-se também, que em média, a eficiência de previsão dos valores de DEPïs para um ano futuro é próxima de 80 por cento.

A Bayesian inference of autoregressive panel data model was applied to real data of Nelore sires Expected Progenie Difference (EPD) during a five-year period (2000-2005). The exact likelihood function and predictive distributions of future observations were considered. The results indicated the importance of sires grouping in homogeneous groups according to accuracy and showed forecast efficiency for EPD values in a future year around 80 percent.

Antiinflammatory and antiulcer properties of tannins from Myracrodruon urundeuva Allemão (Anacardiaceae) in rodents.

Myracrodruon urundeuva Allemão is a plant utilized in Northeast Brazil as an antiinflammatory, wound healing and in gynecological illnesses. The objectives of the present study were to evaluate the antiinflammatory and antiulcer properties of the tannin-enriched fraction (TEF) isolated from the stem bark of M. urundeuva, in the formalin test, in mice, and in carrageenan-induced paw edema and gastric ulcer models, in rats. The results showed that TEF dose-dependently inhibited both phases of the formalin test. However, the effect was predominant in the 2nd phase of the response where inhibitions of 47%, 76% and 85% were observed, with doses of 5, 10 and 50 mg/kg, i.p. In the carrageenan-induced paw edema, significant inhibitions were observed at 3 h (44%) and 4 h (28%), with a dose of 10 mg/kg, i.p. TEF also significantly decreased by 37%, 43% and 57% gastric ulceration induced by indomethacin, at doses of 10, 20 and 50 mg/kg p.o. In the ethanol-induced gastric ulcer model, TEF was less effective, and significant inhibitions (42% to 46%) were observed only with doses of 100 and 200 mg/kg, p.o., respectively. In conclusion, it was shown that TEF presents antiinflammatory and antiulcer effects, partly due to its antioxidant action, known to be present in polyphenols, including tannins.

Experimental Trypanosoma evansi infection in donkeys: hematological, biochemical and histopathological changes

Five adult donkeys were experimentally infected with Brazilian strain of Trypanosoma evansi originally isolated from a naturally infected dog to study the hematological biochemical and histopathological alterations during the evolution of the disease. The course of the experimental infection was followed up to 145 days. Hematological analyses of the infected donkeys revealed a marked decline in hemoglobin, packed-cell volume, and erythrocyte count. Anemia was observed after successive peaks of parasitemia. Biochemical analyses showed increased levels of icterus index, serum globulins and decreased serum albumin and glucose values. All infected donkeys revealed enlargement of spleen and its white pulp, enlargement of mediastinal lymph nodes and lungs congestion. The main histopathological features consisted of meningoencephalitis. Demyelination in some areas of the cerebellum pediculus and neuropil vacuolization were observed. This study showed that donkeys infected with a Brazilian strain of T. evansi developed a chronic disease.

Cinco jumentos, adultos foram infectados experimentalmente com cepa brasileira de Trypanosoma evansi, isolada de um cão naturalmente infectado, com o intuito de observar as alterações hematológicas, bioquímicas e histopatológicas durante a evolução da enfermidade. O curso da infecção experimental foi de 145 dias. Análise hematológica dos jumentos infectados revelou declínio nos valores de hemoglobina, hematócrito e contagem total de eritrócitos. Notou-se anemia após sucessivos picos de parasitemia. Análise bioquímica indicou aumento dos níveis de índice ictérico, globulinas séricas e diminuição dos valores séricos de albumina e glicose. Todos os jumentos infectados apresentaram aumento do baço e de sua polpa branca, aumento de linfonodos mediastínicos e congestão pulmonar. Meningoencefalite foi o principal achado histopatológico. Em algumas áreas do pedículo cerebelar foram observadas desmielinização, além de vacuolização do neurópilo. O estudo mostrou que jumentos infectados com a cepa brasileira do T. evansi desenvolveram doença crônica.

Análise Bayesiana da curva de crescimento de cordeiros da raça Santa Inês / Bayesian analysis of growth curve of Santa Inês lambs

The logistic growth function was fitted to Santa Inês lamb through the Bayesian methodology. The absolute growth rates were taken in each time interval. The methodology was efficient, generating expressive values for parameters estimates and allowed the interval estimation for absolute growth rates.

Experimental Trypanosoma evansi infection in South American coati (Nasua nasua): hematological, biochemical and histopathological changes.

The course of an experimental Trypanosoma evansi infection in coatis (Nasua nasua, carnivora, Procyonidae) was followed for 262 days. Hematological analysis of the infected coatis revealed a marked decline in hemoglobin, packed-cell volume, and total erythrocyte count. An intense anemia followed the first wave of parasitemia and persisted until the end of the experimental period. Biochemical analysis showed increased serum levels of alanine aminotransferase and aspartate aminotransferase and decreased albumin. The main histopathological features consisted of myocarditis with the presence of degenerate cardiac fibers and meningoencephalitis. This study has shown that coatis infected with T. evansi develop a chronic disease.