Andrographolide Inhibits Expression of NLPR3 Inflammasome in Canine Mononuclear Leukocytes.

Inflammasomes are multiprotein complexes that trigger processes through caspase-1 activation, leading to the maturation of proinflammatory cytokines, such as IL-1ß and IL-18. The gene encoding the inflammasome stimulatory protein NLRP3 is conserved in canines. Caspase-1/4 homologues have been identified in multiple carnivores, including canines, and caspase-1 activity has been shown in humans. The NLRP3 inflammasome has also been described in some canine inflammatory diseases. Andrographolide, a labdane diterpene, is the principal active ingredient in the herb Andrographis paniculate. The objective of this study was to determine the effect of andrographolide on the gene expression of the components of the NLRP3 inflammasome, proinflammatory cytokines, and IL-1ß secretion in canine peripheral blood mononuclear cells. For this, MTT assays and real-time PCR were employed to assess the cytotoxicity and gene expression. Further, an ELISA test was performed to measure the IL-1ß concentration. The findings reveal that andrographolide significantly reduces the expression of NLRP3, caspase-1/4, IL-1ß, and IL-18. Additionally, it decreases the secretion of IL-1ß and other proinflammatory cytokines, including IL-6, IL-8, and TNF-α. The results show that andrographolide decreases the expression of NLRP3, caspase-1/4, IL-1ß, and IL-18. Andrographolide also reduces proinflammatory cytokines expression, and decreases IL-1ß secretion. This indicates that andrographolide can interfere with the activation and function of the inflammasome, resulting in a decrease in the inflammatory response in canines. Research in this area is still budding, and more studies are necessary to fully understand andrographolide's mechanisms of action and its therapeutic potential in relation to the NLRP3 inflammasome in dogs.

Exploring the role of healthy dogs as hosts of enterohepatic Helicobacter species using cultivation-dependent and -independent approaches.

Enterohepatic Helicobacter (EHH) species have been increasingly associated with acute gastroenteritis, inflammatory bowel disease and hepatobiliary diseases in humans. However, their host range and transmission routes are poorly understood. Therefore, the aim of this study was to determine the presence of EHH in healthy dogs using both cultivation-dependent and -independent methods. Three hundred and ninety faecal samples from domestic dogs without gastrointestinal symptoms were analysed between June 2018 and July 2019 in Valdivia (South of Chile). Samples were inoculated on selective medium and in parallel were filtrated over an antibiotic-free blood agar. Both media were incubated in a microaerobic atmosphere at 37°C for 7 days. Colonies were identified by PCR and phylogenetic analysis. A subset of 50 samples (half of them positive for EHH by cultivation and the remaining half negative) was analysed by PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) for direct detection. Cultivation method detected EHH in 15.4% (60/390) of the samples, being the most prevalent species H. canis (5.8%, 23/390) and H. canicola (5.1%, 20/390), followed by H. bilis (3.6%, 14/390) and 'H. winghamensis' (1.3%, 5/390). In contrast, PCR-DGGE method detected Helicobacter DNA in almost all (96%, 48/50) tested samples. On the other hand, the method used also allowed to isolate other Campylobacterales, in fact 44.3% (173/390) of the samples were positive for Campylobacter upsaliensis (43.3%, 169/390) followed by C. jejuni (2.0%, 8/390). Moreover, two strains that presented Campylobacter-like morphology were finally identified as Anaerobiospirillum succiniciproducens. Our results indicate that healthy domestic dogs commonly carry EHH and other Campylobacter species. However, further studies are needed to determine whether and how these Helicobacter and Campylobacter species can be transmitted to humans.

Effect of Type of Pregnancy on Transcriptional and Plasma Metabolic Response in Sheep and Its Further Effect on Progeny Lambs.

The following study was performed in order to determine the effect of type of pregnancy on the transcriptional expression of genes that are engaged in angiogenesis and cell turnover/lactogenesis in the ewe mammary gland, evaluating its impact on the plasma metabolic response. In addition, an assessment of its further influence on plasma metabolic response, performance, and muscle transcriptional expression of lipogenic enzymes in progeny lambs was made. Thirteen Ile de France sheep (six twin- and seven single-bearing ewes) were allocated to graze ad libitum naturalized pasture from d 45 pre-partum to day 70 post-partum, while keeping their lambs on the same diet until day 60 after weaning. The samples were collected at different times and analyzed by qRT-PCR and plasma metabolic indicators. The data were processed using SPSS package. The results showed that twin-bearing ewes overexpressed VEGFR1 at birth, and BCL2 at birth and day 35 post-partum; however, single-bearing ewes overexpressed CAIV and IGF1 at day 35 post-partum. Similar metabolite concentrations in blood plasma were found between groups of ewes. The plasma metabolic response in lambs was similar between groups and it did not influence their performance, where a similar transcriptional expression of lipogenic enzymes in muscle was observed. Therefore, the type of pregnancy can explain the slight differences in mRNA expression that were found in angiogenesis and cell turnover/lactogenesis in mammary gland, although these differences not only did not affect the plasma metabolic response in ewes, but they also had no influence on plasma metabolic response, performance, and muscle transcriptional expression of their lambs.

Intra Vitam Diagnosis of Neglected Gurltia paralysans Infections in Domestic Cats (Felis catus) by a Commercial Serology Test for Canine Angiostrongylosis and Insights into Clinical and Histopathological Findings-Four-Case Report.

Gurltia paralysans is a metastrongyloid nematode which belongs to the Angiostrongylidae family and presents tropism for veins of the subarachnoid space in vivo of domestic and wild felids causing a progressive and chronic clinical manifestation of paraparesis/paraplegia. The geographic distribution of G. paralysans includes rural and periurban areas of South America and was recently reported in Europe. To date, a definitive diagnosis of feline gurltiosis is still conducted by post-mortem examination of the spinal cord in affected animals. A presumptive diagnosis of feline gurltiosis can also be achieved based on data of compatible clinical signs and associated epidemiological risk factors. The aim of this preliminary study was to evaluate the commercial serological test Angio Detect TM® (IDEXX Laboratories) as a possible diagnostic method of feline gurltiosis in vivo. For the study, 10 domestic felines (Felis catus) which originated from a high endemic area of Southern Chile, were analyzed. All felines presented chronic paraparesis or severe paraplegia. Subsequently, commercial Angio Detect TM® serological tests for the detection of closely related Angiostrongylus vasorum in canids were performed according to manufacturer's instructions. Conducted serological tests were positive in seven of ten felines showing paraplegia/paraparesis and presumably infected with G. paralysans, and four of them were additionally necropsied, and presented macroscopic findings compatible with feline gurltiosis. Furthermore, the presence of adult female and male G. paralysans specimens at the level of the subarachnoid vasculature in affected spinal cord segments were observed during necropsy. Histopathology demonstrated severe eosinophilic meningomyelitis, coagulopathies with thrombosis in G. paralysans-parasitized leptomeningeal veins. Results of this preliminary study suggest a cross-reaction between A. vasorum-specific antigens, which also parasitize blood vessels in vivo, and G. paralysans when using an Angio Detect TM® test, which suggests its helpful use as a new diagnostic method for feline gurltiosis in live domestic felines. Additional specific antigen research will be required in order to better understand the sensitivity and specificity of A. vasorum antigens used in this test and for existing cross-reactivity with G. paralysans-derived antigens for future a suitable intra vitam immunodiagnosis of neglected feline gurltiosis.

Vincristine, carboplatin and cisplatin increase oxidative burst induced by PAF in canine neutrophils.

Myelosupression resulting from chemotherapy has been widely described in veterinary medicine; however, there is limited information relating to alterations in neutrophil function after chemotherapy in dogs with cancer. The aim of this study was to determine the non-proliferative effects of vincristine, carboplatin, and cisplatin on canine neutrophils by evaluating activation of oxidative and non-oxidative responses. Neutrophils were isolated from venous blood. Levels of reactive oxygen species (ROS) and metalloproteinase 9 (MMP-9) were measured in vitro during neutrophil exposure to these chemotherapeutic agents for 15 min followed by stimulation with platelet activating factor (PAF). ROS production was detected via luminescence, and MMP- 9 liberation was determined by zymography. The chemotherapeutic agents caused an increase in PAF-induced ROS production, but no change in the non-oxidative response was observed. These results suggest that these chemotherapeutic agents may act as priming agents by increasing the oxidative response. These effects could be beneficial for dogs with cancer by supporting their immune systems; however, excessive ROS liberation has been associated with inflammation, neutrophil-mediated cell injury, carcinogenesis, and metastasis. Clinical studies are necessary to evaluate the significance of these findings.

Evidence of interspecies transmission of pathogenic Leptospira between livestock and a domestic cat dwelling in a dairy cattle farm.

A domestic cat dwelling in a dairy cattle farm with haematuria was referred for a physical examination. The examination showed no abnormalities therefore complementary exams were performed. Leukocytosis with neutrophilia, monocytosis and hyperproteinaemia were detected. The urine analysis showed a bacterial infection without ultrasound findings. Serological titers to Leptospira interrogans serovar Pomona and Autumnalis were detected. Molecular analysis demonstrated the presence of Leptospira spp. in urine. The findings were consistent with subclinical leptospirosis. The cattle herd had evidence of Leptospira infection. The microbiological exams confirmed the presence of the Leptospira spp. in urine and serum. According to the evidence presented in this study, cats that dwell within a dairy farm could play a role in the Leptospira infection epidemiologically. The importance of feline leptospirosis must be evaluated with leptospirosis control in livestock.

Computer tomographic imaging in 4 dogs with primary nasal canine transmissible venereal tumor and differing cellular phenotype.

Primary nasal canine transmissible venereal tumor (CTVT) without genital affection is uncommon. The aim of this report was to describe the primary nasal CTVT findings and CT staging in 4 dogs with different cytological phenotypes. Three male dogs and 1 bitch were evaluated for their chronic histories of sneezing, snoring, mucopurulent nasal discharge and nasal deformation. Cytological examination of nasal secretions suggested CTVT, confirmed by histopathological examination and LINE-1/c-myc. Males had the plasmacytoid phenotype of CTVT, and the bitch had the lymphocytoid phenotype. CTVT were staged based on the CT findings using modified Adams staging system. The bitch was classified as stage 1, 2 males were classified as stage 3 and 1 male as stage 4. All dogs had a complete tumoral remission after chemotherapy. Plasmacytoid phenotype was identified in cases with most important damage of the nasal cavity. However, the cytological type did not affect the response to chemotherapy.

Aerogels made of chitosan and chondroitin sulfate at high degree of neutralization: Biological properties toward wound healing.

In this study, highly neutralized, highly porous, and ultralight polymeric aerogels prepared from aqueous colloidal suspensions of chitosan (CS) and chondroitin sulfate (ChS) nanocomplexes, formulated as quasi-equimolar amounts of both, are described. These aerogels were designed as healing agents under the inspiration of minimizing the amount of matter applied to wounds, reducing the electrostatic potential of the material and avoiding covalent cross-linkers in order to decrease metabolic stress over wounds. Aerogels synthesized under these criteria are biocompatible and provide specific properties for the induction of wound healing. They do not affect neither the metabolic activity of cultured 3T3 fibroblasts nor the biochemical parameters of experimental animals, open wounds close significantly faster and, unlike control wounds, complete reepithelialization and scarring can be attained 14 days after surgery. Because of its hydration abilities, rapid adaptation to the wound bed and the early accelerator effect of wound closure, the CS/ChS aerogels appear to be functional inducers of the healing. Previous information show that CS/ChS aerogels improve wound bed quality, increase granulation tissue and have pain suppressive effect. CS/ChS aerogels are useful as safe, inexpensive and easy to handle materials for topical applications, such as skin chronic wounds. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2464-2471, 2018.

Fatty acid extraction from sewage sludge using a porous polyethylene sorbent obtained from agricultural covers.

Fatty acids from a sewage sludge of a wastewater treatment plant were extracted using a porous polyethylene (pPE) material prepared using agricultural plastic films. The pPE fatty acid extraction profile and yield from the sewage sludge was compared to toluene as extraction solvent. The chemical composition of the extracted fatty acids were the same in both cases, but when the extraction yield was normalized by the used mass of pPE or toluene, pPE resulted six times more efficient than toluene. The pPE bulk density and porosity obtained using different biodiesel:vegetable oil mixtures ranged between 0.24 g cm-3-0.40 g cm-3 and 58-74%, respectively. Critical surface tension of the pPE was (26.9 ±â€¯1.8) dyn cm-1 while for the original polyethylene film it was (19.9 ±â€¯3.1) dyn cm-1. Biodiesel and oleic acid were used as models of hydrophobic substances for extraction studies. pPE Maximum sorption capacities were: 6.8 g biodiesel g-1 pPE and 9.7 g oleic acid g-1 pPE. Sorption capacity remained practically constant after ten biodiesel sorption/extraction cycles. Pesticide (trifluralin) residues in the pPE were analyzed to assure that xenobiotic presence can be removed during the manufacturing process, finding that less than 0.84% of an initially present pesticide remained in the pPE.

Gangrena de Fournier / Fournier's gangrene

Se presenta el caso de un paciente con gangrena de Fournier o fascitis necrosante. Después de su ingreso en la Unidad de Cuidados Intensivos se le realizó tratamiento quirúrgico precoz (incisión y drenaje amplio de toda la zona afectada así como desbridamiento de la zona de necrosis); se tomó muestra de pus para estudio microbiológico y se comenzó tratamiento antibiótico de inmediato (ceftriaxona, amikacina y metronidazol). Se identificaron como agentes causantes del proceso Pseudomonas aeruginosa y Escherichia coli (sensibles a la ceftriaxona y la amikacina). Como tratamiento coadyuvante, se aplicaron 10 sesiones de cámara hiperbárica y 12 dosis de factor de crecimiento humano recombinante liofilizado, que se infiltraron en el interior de la zona quirúrgica para contribuir al cierre por tercera intención(AU)

This is the case of a patient presenting with Fournier gangrene of necrotizing fasciitis. After admission in Intensive Care Unit (ICU) he undergoes an early surgical treatment (incision and wide drainage of the whole involved zone as well as debridement of necrosis zone); a pus sample was took for microbiologic study and a immediate antibiotic treatment was started (Ceftriaxone, Amikacin and Metronidazole). Pseudomonas aeruginosa and Escherichia coli (sensitive to Ceftriaxone and Amikacin) were identified as the causative agents of process. As adjuvant treatment ten sessions of hyperbaric chamber were applied and twelve dose of lyophilized recombinant human growth factor with infiltration into the surgical zone for the third intention closure(AU)

Miomatosis uterina gigante / Giant uterine myomatosis

El gran tamaño de algunos tumores ha sido descrito con asombro a través del tiempo. Entre ellos se incluyen los ginecológicos, sobre los cuales se describen casos de crecimiento descomunal, e incluyen los uterinos u ováricos. El caso de mayor tamaño fue descrito por Hunt en 1888 como hallazgo en una necropsia, con el "asombroso" peso de 63,5 kg. La terminología de estos tumores de gran tamaño contiene calificativos muy variados y confusos, entre los cuales se incluyen "inmensos", "enormes" "extensos", "voluminosos", "masivos", "grandes", "muy grandes", "gigantes", "gigantescos". Beacham y otros, en 1971 definieron como "gigantes" a los tumores uterinos u ováricos con peso superior a 25 lbs (11,3 kg). Briceño-Pérez y otros en el año 2001 propusieron utilizar el término de "grandes miomas uterinos" (GMU) para aquellos con peso entre 0,8 Kg (peso límite aproximado para decidir la vía quirúrgica abdominal o vaginal) y 11,3 Kg (peso límite establecido por Beacham y colaboradores para los miomas uterinos gigantes). El caso que se presenta fue operado el día 20 de enero de 2009 en el Centro de Investigaciones Médico Quirúrgicas, al cual se le realizó histerectomía total con el diagnóstico de miomatosis gigante del útero cuyo peso fue de 11,31 kg. Se hacen recomendaciones para el manejo quirúrgico de las pacientes con tumores ginecológicos gigantes.

The large size of some tumors has been described with surprise over the time. Among them are included the gynecologic ones, on which are presented cases of an enormous growth and the uterine and ovarian types are also included. The largest size of a tumor was described by Hunt in 1888 as a necropsy finding, which weighing 63,5 kg. Terminology of these large size tumors include very varied and confused epithets: "inmenses", "huges", "extensive", "bulkies", "mass", "larges", "very larges", "giants" , and "gigantics". Beachman et al, in 1971 defined as "giants" the uterine or ovarian tumors weighing more than 25 pounds (11,3 kg). Briceño-Pérez et al in 2001 proposed the use of the term "large uterine myomas" (LUM) for those weighing between 0,8 kg (limit weight established by Beacham et al for the giant uterine myomas). Present case was operated on in January 20, 2009 in Medical and Surgical Researches Center, performing a total hysterectomy diagnosed with uterine giant myomatosis weighing 11,31 kg. Authors made recommendations on surgical management of patients presenting giant gynecological tumors.

Analytical method for assessing potential dermal exposure to captan, using whole body dosimetry, in small vegetable production units in Argentina.

An analytical method has been developed that can be used to determine the potential dermal exposure (PDE) of workers to the pesticide captan in small-scale horticultural production units. The methodology is based on the whole body dosimetry technique, using a cotton coverall and cotton gloves as sampling media, with protective clothing worn beneath the cotton media to protect the operator. The quantitative determination of captan was done by gas chromatography-electron capture detector (GC-ECD), with the analytical method validated by measuring limits of detection and quantification, linear ranges, sample recovery and precision. Special emphasis is placed on factors that affected the stability of captan during chromatographic determination. The data generated for potential dermal exposure are presented separately for mixing/loading and application activities. These data are compared with values obtained with visible tracers using a similar field technique. Margin of safety (MOS) values are also calculated for the agricultural procedures studied.