Numerous studies on endometritis of postpartum dairy cows describe the bacterial aetiology of the condition. However, studies on fungal agents causing inflammatory conditions of the reproductive tract are limited. The objective of the present study was to identify the mycotic isolates from the reproductive tract of postpartum dairy cows and analyse their susceptibility to antifungal agents. In this study, 70 endometrial cytobrush samples were collected from dairy cows 40 - 120 days postpartum (dpp) and cultured in Sabouraud dextrose agar (SDA). The isolated fungal cultures were identified by Lactophenol Cotton Blue staining and were subjected to antifungal susceptibility testing. The frequency of isolation of fungi from endometrial samples was 15.71 per cent (11/70). Aspergillus niger was isolated from four (33.33%) samples, followed by Penicillium spp. (25%), Aspergillus fumigatus (16.67%), Mucor spp. (16.67%) and Absidia spp. (8.33%). Antifungal susceptibility testing revealed that 50 per cent of the isolates were susceptible to fluconazole, whereas only 25 per cent of the isolates were completely resistant to amphotericin-B. The prevalence of fungal infection in reproductive tract was higher in cows with an age of ≥ 5 years and a parity of ≥ 2, though this was not significant. A significantly higher number of cattle with history of intrauterine antibiotic therapy were positive for fungal culture. However, the association between prevalence of fungi and reproductive complications was statistically non-significant. Keywords: Postpartum dairy cows, fungal infections, Aspergillus niger, mycotic infections
John M. Kelly, MD, Zinan Hu, BS, Felipe Takaesu, BS
et al.
Objectives: Our goal was to conduct a hemodynamic analysis of a novel animal model of Fontan physiology. Poor late-term outcomes in Fontan patients are believed to arise from Fontan-induced hemodynamics, but the mechanisms remain poorly understood. Recent advances in surgical experimentation have resulted in the development of a chronic sheep model of Fontan physiology; however, detailed analysis of this model is lacking. Methods: We created a single-stage Fontan model in juvenile sheep with normal biventricular circulation. The superior vena cava was anastomosed to the main pulmonary artery, and the inferior vena cava was connected to the main pulmonary artery using an expanded polytetrafluoroethylene conduit. Longitudinal hemodynamics, including catheterization and magnetic resonance imaging were evaluated. Results: Four out of 12 animals survived, with the longest surviving animal living 3 years after single-stage Fontan. We showed a significant era effect regarding survival (1 out of 8 and subsequently 3 out of 4 animals surviving beyond 2 months) attributed in large part to the procedural learning curve. Key characteristics of Fontan hemodynamics, namely systemic venous hypertension and low normal cardiac output, were observed. However, recapitulation of passive human Fontan hemodynamics is affected by volume loading of the right ventricle given an anatomic difference in sheep azygous venous anatomy draining to the coronary sinus. Conclusions: A significant learning curve exists to ensure long-term survival and future surgical modifications, including banding of the main pulmonary artery and ligation of the azygous to coronary sinus connection are promising strategies to improve the fidelity of model hemodynamics.
Diseases of the circulatory (Cardiovascular) system, Surgery
Spent Japanese quail (SJQ) meat was tenderized with ginger extract and utilized in the preparation of tandoori. The ginger extract level for tenderization of SJQ meat was optimized by physico-chemical and sensory analyses. The SJQ meat was treated with two levels of ginger extract and used for tandoori preparation along with two controls (broiler quail meat and spent quail meat). Japanese quail tandoori was prepared as four treatments namely CYB (Control young broiler), CSA (Control spent adult), G20 (Spent adult - ginger treatment 20%) and G40 (Spent adult - ginger treatment 40%). The aqueous ginger extract was prepared from fresh ginger rhizome (Zingiber officinale Roscoe). The SJQ was marinated in ginger extract marinade, tumbled, and placed in the chiller (4±1⁰C) for 24 hours and utilized for tandoori preparation. The cooked tandoori of all the treatments were analysed for the physico-chemical and sensory parameters. With ginger extract marination, the pH of cooked tandoori was not affected and the product yield was improved. In the organoleptic evaluation, the appearance score increased in the treatment with 20% ginger extract. The flavour score was significantly (p<0.05) higher in CYB and G20. The SJQ was significantly (p<0.05) tenderized by treatment with 20% and 40% ginger extract. CYB and G20 had significantly (p<0.01) higher scores for overall acceptability scores. It is concluded that 20% ginger extract level could effectively be used to tenderize the SJQ meat for the preparation of tandoori without affecting the physico-chemical and sensory properties.
Ebanja Joseph Ebwanga, Stephen Mbigha Ghogomu, Jan Paeshuyse
African swine fever and swine erysipelas are two devastating diseases with similar manifestations ravaging the domestic pig industry. Only a single phylogenetic study has been carried out in Cameroon, and neither an extensive genotyping aimed at identifying the different serotypes nor has an appropriate differential diagnosis of different species of <i>Erysipelothrix</i> has been effected in ASF-infected animals. Of the 377 blood or tissue samples randomly collected from pig farms and slaughter slabs from January to August 2020, 120 were positive for ASFV (by PCR), giving a prevalence of 31.83%. Intragenomic resolution through sequencing divulged the presence of genotypes I, and Ia, two variants with 19 (ABNAAAACBNABTDBNAFA) and six (ABNAFA) tandem repeat sequences (TRS), serotype IV, and a single GGAATATATA repeat. The sole presence of <i>E. tonsillarum</i> (avirulent species) and not <i>E. rhusiopathiae</i> (virulent species) indicates that the severity observed during the 2020 ASF outbreak in the sampled regions was exclusively due to ASFV genotype I infection. Such characterisations are necessary for designing effective control measures and future potential vaccine candidates.
Sandra Díaz del Moral, Silvia Barrena, Francisco Hernández-Torres
et al.
Expression of Wilms’ tumor suppressor transcription factor (WT1) in the embryonic epicardium is essential for cardiac development, but its myocardial expression is little known. We have found that WT1 is expressed at low levels in 20–25% of the embryonic cardiomyocytes. Conditional ablation of WT1 using a cardiac troponin T driver (Tnnt2Cre) caused abnormal sinus venosus and atrium development, lack of pectinate muscles, thin ventricular myocardium and, in some cases, interventricular septum and cardiac wall defects, ventricular diverticula and aneurisms. Coronary development was normal and there was not embryonic lethality, although survival of adult mutant mice was reduced probably due to perinatal mortality. Adult mutant mice showed electrocardiographic anomalies, including increased RR and QRS intervals, and decreased PR intervals. RNASeq analysis identified differential expression of 137 genes in the E13.5 mutant heart as compared to controls. GO functional enrichment analysis suggested that both calcium ion regulation and modulation of potassium channels are deeply altered in the mutant myocardium. In summary, together with its essential function in the embryonic epicardium, myocardial WT1 expression is also required for normal cardiac development.
Kenneth Ssekatawa, Kenneth Ssekatawa, Denis K. Byarugaba
et al.
Antibiotics have been the nucleus of chemotherapy since their discovery and introduction into the healthcare system in the 1940s. They are routinely used to treat bacterial infections and to prevent infections in patients with compromised immune systems and enhancing growth in livestock. However, resistance to last-resort antibiotics used in the treatment of multidrug-resistant infections has been reported worldwide. Therefore, this study aimed to evaluate green synthesized nanomaterials such as silver nanoparticles (AgNPs) as alternatives to antibiotics. UV-vis spectroscopy surface plasmon resonance peaks for AgNPs were obtained between 417 and 475 nm. An X-ray diffraction analysis generated four peaks for both Prunus africana extract (PAE) and Camellia sinensis extract (CSE) biosynthesized AgNPs positioned at 2θ angles of 38.2°, 44.4°, 64.5°, and 77.4° corresponding to crystal planes (111), (200), (220), and (311), respectively. A dynamic light-scattering analysis registered the mean zeta potential of +6.3 mV and +0.9 mV for PAE and CSE biosynthesized nanoparticles, respectively. Fourier transform infrared spectroscopy spectra exhibited bands corresponding to different organic functional groups confirming the capping of AgNPs by PAE and CSE phytochemicals. Field emission scanning electron microscopy imaging showed that AgNPs were spherical with average size distribution ranging from 10 to 19 nm. Biosynthesized AgNPs exhibited maximum growth inhibitory zones of 21 mm with minimum inhibitory concentration and minimum bactericidal concentration of 125 and 250 μg/ml, respectively, against carbapenem-resistant bacteria.
Abstract The cyclic GMP-AMP (cGAMP) synthase (cGAS) plays a critical role in host defense by sensing cytosolic DNA derived from microbial pathogens or mis-located cellular DNA. Upon DNA binding, cGAS utilizes GTP and ATP as substrates to synthesize cGAMP, leading to MITA-mediated innate immune response. In this study, we identified the phosphatase PPP6C as a negative regulator of cGAS-mediated innate immune response. PPP6C is constitutively associated with cGAS in un-stimulated cells. DNA virus infection causes rapid disassociation of PPP6C from cGAS, resulting in phosphorylation of human cGAS S435 or mouse cGAS S420 in its catalytic pocket. Mutation of this serine residue of cGAS impairs its ability to synthesize cGAMP upon DNA virus infection. In vitro experiments indicate that S420-phosphorylated mcGAS has higher affinity to GTP and enzymatic activity. PPP6C-deficiency promotes innate immune response to DNA virus in various cells. Our findings suggest that PPP6C-mediated dephosphorylation of a catalytic pocket serine residue of cGAS impairs its substrate binding activity and innate immune response, which provides a mechanism for keeping the DNA sensor cGAS inactive in the absence of infection to avoid autoimmune response.
The aim of this work was the in vitro expression of the recombinant fusion (F) protein of Newcastle disease virus (NDV). The pBT7-N-His-Fusion-NDV expression plasmid which carries the recombinant F protein encoding gene from local Indonesian isolates, was prepared and transformed into E. coli BL21 (DE3). To detect bacterial colonies carrying the recombinant plasmid, a restriction endonuclease analysis was performed using the EcoRI restriction endonuclease. These results showed that the pBT-N-His-Fusion-NDV plasmid was successfully isolated with a size of 4.601 bp, and three recombinant plasmids carrying the gene coding for the recombinant F protein of NDV were obtained. Selected recombinant plasmids were then in vitro by using a cell-free protein expression system followed by visualization of the recombinant F protein on a 12% SDS-PAGE gel both by Coomassie Brilliant Blue staining and Western blotting. Recombinant F protein was successfully in vitro expressed by using a cell-free protein expression system as indicated by a specific single protein band with a molecular mass of 25.6 kDa.
Feline panleukopenia is a rapidly progressive viral disease of domestic cats caused by a single stranded parvovirus of the genus Protoparvovirus. The virus, owing to its ubiquitous nature, remains persistent in the environment for extended periods leading to frequent exposures in naïve cat population. Clinically, the disease is manifested as haemorrhagic enteritis characterised by vomiting, diarrhoea, dysentery, dehydration, shock and death. Previous studies conducted by Parthiban et al. (2014) and Koulath et al. (2017) detected feline panleukopenia in domestic cats of India using polymerase chain reaction (PCR) targeting VP2 gene of the virus. However, data related to the clinical manifestations of this disease in cats in India is limited. The current paper
deals with the clinical findings of cats affected with feline panleukopenia in Kerala.
Patiwat Kongdang, Chatchadawalai Chokchaitaweesuk, Siriwan Tangyuenyong
et al.
Combinations of IL-1β and other proinflammatory cytokines reportedly promote the severity of arthritis. We aimed to investigate the effects of IL-1β combined with IL-17A on cartilage degradation and synthesis in in vitro models. Cartilage explant degradation was determined using sulfated glycosaminoglycans (S-GAGs) levels, matrix metalloproteinase (MMP13) gene expression, uronic acid, and collagen contents. Cell morphology and accumulation of proteoglycans were evaluated using hematoxylin-eosin and safranin O staining, respectively. In the pellet culture model, expressions of cartilage-specific anabolic and catabolic genes were evaluated using real-time qRT-PCR. Early induction of MMP13 gene expression was found concomitantly with significant S-GAGs release. During the prolonged period, S-GAGs release was significantly elevated, while MMP-13 enzyme levels were persistently increased together with the reduction of the cartilaginous matrix molecules. The pellet culture showed anabolic gene downregulation, while expression of the proinflammatory cytokines, mediators, and MMP13 genes were elevated. After cytokine removal, these effects were restored to nearly basal levels. This study provides evidence that IL-1β combined with IL-17A promoted chronic inflammatory arthritis by activating the catabolic processes accompanied with the suppression of cartilage anabolism. These suggest that further applications, which suppress inflammatory enhancers, especially IL-17A, should be considered as a target for arthritis research and therapy.
Festus A. Adejoro, Abubeker Hassen, Mapitsi S. Thantsha
Objective The use of tannin extract and other phytochemicals as dietary additives in ruminants is becoming more popular due to their wide biological actions such as in methane mitigation, bypass of dietary protein, intestinal nematode control, among other uses. Unfortunately, some have strong astringency, low stability and bioavailability, and negatively affecting dry matter intake and digestibility. To circumvent these drawbacks, an effective delivery system may offer a promising approach to administer these extracts to the site where they are required. The objectives of this study were to encapsulate acacia tannin extract (ATE) with native starch and maltodextrin-gum arabic and to test the effect of encapsulation parameters on encapsulation efficiency, yield and morphology of the microparticles obtained as well as the effect on rumen in vitro gas production. Methods The ATE was encapsulated with the wall materials, and the morphological features of freeze-dried microparticles were evaluated by scanning electron microscopy. The in vitro release pattern of microparticles in acetate buffer, simulating the rumen, and its effect on in vitro gas production was evaluated. Results The morphological features revealed that maltodextrin/gum-arabic microparticles were irregular shaped, glossy and smaller, compared with those encapsulated with native starch, which were bigger, and more homogenous. Maltodextrin-gum arabic could be used up to 30% loading concentration compared with starch, which could not hold the core material beyond 15% loading capacity. Encapsulation efficiency ranged from 27.7%±6.4% to 48.8%±5.5% in starch and 56.1%±4.9% to 64.8%±2.8% in maltodextrin-gum arabic microparticles. Only a slight reduction in methane emission was recorded in encapsulated microparticles when compared with the samples containing only wall materials. Conclusion Both encapsulated products exhibited the burst release pattern under the pH conditions and methane reduction associated with tannin was marginal. This is attributable to small loading percentages and therefore, other wall materials or encapsulation methods should be investigated.
Annisa Rahmah Furqaani, Sri Redjeki, Dwirini Retno Gunarti
Physical exercise can enhance tryptophan transport into the brain so that it will also increase serotonin levels in the brain. Therefore, it may influence many brain functions, such as learning and memory. This study aimed to determine the effect of physical exercise on spatial learning and serotonin levels in the brain of adult male Wistar rats. Biochemistry Laboratory of Department of Biochemistry & Molecular Biology, Faculty of Medicine, Universitas Indonesia was the study place which conducted in January–April 2013. Sixteen adult male rats randomly divided into two groups, the control group, and the treatment group. Physical exercise for the treatment group for four weeks using the animal treadmill at 15 m/min in speed for 15 minutes in the 1st week and 25 minutes for the next three weeks. Learning and memory test using water-E maze apparatus once a week. At the end of the exercised period, animals were sacrificed, and the brains were isolated. The measurement of serotonin and tryptophan levels was done using high-performance liquid chromatography (HPLC). The results showed that physical exercise improved animals performance in learning and memory test, exercised group made fewer errors at third and fourth week (p<0.05). Serotonin levels in the brain of exercised group was significantly higher than that in control group (p<0.05). These results indicated that the enhancement of serotonin levels in the brain induced by physical exercise is involved in improving spatial learning and memory.
PENGARUH LATIHAN FISIK TERHADAP KEMAMPUAN BELAJAR SPASIAL DAN KADAR SEROTONIN PADA OTAK TIKUS DEWASA
Latihan fisik diketahui dapat meningkatkan transpor triptofan melewati sawar otak sehingga dapat meningkatkan kadar serotonin di otak. Oleh karena itu, latihan fisik berperan memengaruhi berbagai fungsi otak termasuk proses belajar dan memori. Penelitian ini bertujuan mengetahui pengaruh latihan fisik aerobik dengan intensitas yang ringan terhadap kemampuan belajar spasial serta kadar serotonin pada otak tikus Wistar dewasa. Penelitian ini dilakukan di Laboratorium Biokimia, Departemen Biokimia & Biologi Molekuler, Fakultas Kedokteran, Universitas Indonesia, Jakarta periode Januari–April 2013. Enam belas ekor tikus jantan dewasa dibagi secara acak menjadi dua kelompok, yaitu kelompok kontrol dan kelompok perlakuan. Latihan fisik diberikan kepada kelompok perlakuan selama 4 minggu menggunakan animal treadmill dengan kecepatan 15 m/menit selama 15 menit pada minggu pertama dan 25 menit pada 3 minggu berikutnya. Uji belajar dan memori dengan perangkat water-E maze dilakukan satu kali/minggu. Setelah masa latihan fisik selesai, hewan coba dikorbankan dan jaringan otak diisolasi. Pengukuran kadar serotonin dan triptofan pada otak dilakukan menggunakan kromatografi cair kinerja tinggi (KCKT). Hasil penelitian menunjukkan bahwa jumlah kesalahan yang dilakukan oleh kelompok perlakuan lebih sedikit secara signifikan pada uji belajar dan memori ke-3 dan ke-4 (p<0,05). Kadar serotonin lebih tinggi secara signifikan pada otak kelompok perlakuan (p<0,05). Hasil penelitian ini mengindikasikan bahwa peningkatan kadar serotonin pada otak yang diinduksi oleh latihan fisik aerobik intensitas ringan terlibat dalam meningkatkan kemampuan belajar dan memori spasial.
Isabel L. Pacheco, Nieves Abril, Rafael Zafra
et al.
Abstract The expression of T regulatory cells (Foxp3), regulatory (interleukin [IL]-10 and transforming growth factor beta [TGF-β]) and proinflammatory (tumor necrosis factor alpha [TNF-α] and interleukin [IL]-1β) cytokines was quantified using real time polymerase chain reaction (qRT-PCR) in the liver of sheep during early stages of infection with Fasciola hepatica (1, 3, 9, and 18 days post-infection [dpi]). Portal fibrosis was also evaluated by Masson’s trichrome stain as well as the number of Foxp3+ cells by immunohistochemistry. Animals were divided into three groups: (a) group 1 was immunized with recombinant cathepsin L1 from F. hepatica (FhCL1) in Montanide adjuvant and infected; (b) group 2 was uniquely infected with F. hepatica; and (c) group 3 was the control group, unimmunized and uninfected. An overexpression of regulatory cytokines of groups 1 and 2 was found in all time points tested in comparison with group 3, particularly at 18 dpi. A significant increase of the number of Foxp3+ lymphocytes in groups 1 and 2 was found at 9 and 18 dpi relative to group 3. A progressive increase in portal fibrosis was found in groups 1 and 2 in comparison with group 3. In this regard, group 1 showed smaller areas of fibrosis than group 2. There was a significant positive correlation between Foxp3 and IL-10 expression (by immunohistochemistry and qRT-PCR) just as between portal fibrosis and TGF-β gene expression. The expression of proinflammatory cytokines increased gradually during the experience. These findings suggest the induction of a regulatory phenotype by the parasite that would allow its survival at early stages of the disease when it is more vulnerable.
Yong Sze Ong,1 Latifah Saiful Yazan,1,2 Wei Keat Ng,1 Mustapha M Noordin,3 Sarah Sapuan,1 Jhi Biau Foo,1 Yin Sim Tor1 1Laboratory of Molecular Biomedicine, Institute of Bioscience, 2Department of Biomedical Science, Faculty of Medicine and Health Sciences, 3Department of Pathology and Veterinary Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia Background: Thymoquinone (TQ), the predominant active lipophilic component in Nigella sativa seed oil, has a variety of pharmacological properties such as anticancer activities. However, translation of TQ to clinical phase is still not possible due to its hydrophobic properties. This problem can be solved by encapsulating it in nanoformulations to enhance its pharmacological properties. In our previous study, TQ has been successfully encapsulated in a nanostructured lipid carrier (hereinafter referred to as TQNLC) with excellent physiochemical properties such as high encapsulation efficiency, high drug-loading capacity, particle diameter less than 100 nm, and stability up to 2 years. In vitro studies also proved that TQNLC exhibited antiproliferative activity toward breast and cervical cancer cell lines. However, no toxicity profile related to this formulation has been reported. In this study, we determine and compare the in vivo toxicity of both TQNLC and TQ. Materials and methods: The in vivo toxicity (acute and subacute toxicity) study was carried out by oral administration of TQNLC and TQ to BALB/c mice. Animal survival, body weight, organ weight-to-body weight ratio, hematological profile, biochemistry profile, and histopathological changes were analyzed. Results: In acute toxicity, TQ that is loaded in nanostructured lipid carrier (NLC) was found to be less toxic than pure TQ. It can be concluded that encapsulation of TQ in lipid carrier minimizes the toxicity of the compound. In the subacute toxicity study, oral administration of 100 mg/kg of TQNLC and TQ did not cause mortality to either male or female but resulted in toxicity to the liver. It is postulated that long-term consumption of TQNLC and TQ may cause toxicity to the liver but not to the extent of altering the functions of the organ. For both treatments, the no observed adverse effect level (NOAEL) was found to be 10 mg/kg/d for mice in both sexes. Conclusion: For long-term oral consumption, TQ and TQNLC at a dose of 10 mg/kg is safe in mice and does not exert any toxic effect. The results provide safety information of TQNLC, which would further help researchers in clinical use. Keywords: thymoquinone, nanostructured lipid carrier, toxicity
Jeong-A Lee,1 Mi-Kyung Kim,1 Hee-Jeong Paek,1 Yu-Ri Kim,2 Meyoung-Kon Kim,2 Jong-Kwon Lee,3 Jayoung Jeong,3 Soo-Jin Choi1 1Department of Food Science and Technology, Seoul Women’s University, Seoul, Republic of Korea; 2Department of Biochemistry and Molecular Biology, Korea University Medical School and College, Seoul, Republic of Korea; 3Toxicological Research Division, National Institute of Food and Drug Safety Evaluation, Chungchungbuk–do, Republic of Korea Purpose: The effects of particle size on the tissue distribution and excretion kinetics of silica nanoparticles and their biological fates were investigated following a single oral administration to male and female rats. Methods: Silica nanoparticles of two different sizes (20 nm and 100 nm) were orally administered to male and female rats, respectively. Tissue distribution kinetics, excretion profiles, and fates in tissues were analyzed using elemental analysis and transmission electron microscopy. Results: The differently sized silica nanoparticles mainly distributed to kidneys and liver for 3 days post-administration and, to some extent, to lungs and spleen for 2 days post-administration, regardless of particle size or sex. Transmission electron microscopy and energy dispersive spectroscopy studies in tissues demonstrated almost intact particles in liver, but partially decomposed particles with an irregular morphology were found in kidneys, especially in rats that had been administered 20 nm nanoparticles. Size-dependent excretion kinetics were apparent and the smaller 20 nm particles were found to be more rapidly eliminated than the larger 100 nm particles. Elimination profiles showed 7%–8% of silica nanoparticles were excreted via urine, but most nanoparticles were excreted via feces, regardless of particle size or sex. Conclusion: The kidneys, liver, lungs, and spleen were found to be the target organs of orally-administered silica nanoparticles in rats, and this organ distribution was not affected by particle size or animal sex. In vivo, silica nanoparticles were found to retain their particulate form, although more decomposition was observed in kidneys, especially for 20 nm particles. Urinary and fecal excretion pathways were determined to play roles in the elimination of silica nanoparticles, but 20 nm particles were secreted more rapidly, presumably because they are more easily decomposed. These findings will be of interest to those seeking to predict potential toxicological effects of silica nanoparticles on target organs. Keywords: biological fate, size effect, target organ