Recurrence of hepatitis C virus after treatment with pegylated interferon and direct acting antivirals in Punjab Pakistan / Recorrência do vírus da hepatite C após tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão

Although increased response rates concomitant in hepatitis C virus but relapse after treatment is threatened. Therefore, it is terrible requirement to evaluate the response of Pegylated interferon and direct acting antivirals in Punjab Pakistan. The study was conducted to find the rate of recurrence of HCV infection after treatment with Pegylated Interferon and Direct Acting Antivirals in Punjab Pakistan. This study was conducted at Department of Pathology, Nawaz Sharif Medical College Gujrat, while treatment effects monitored in different Government and Private Hospitals of Punjab, Pakistan. Total 973 patients who administered the recommended dose and divided in two groups (i) Interferon based therapy (ii) direct acting antivirals (DAAs).Other parameters like ALT and viral load studied. The rate of recurrence was higher in female infected with genotype 2b and in male with mixed genotype 3a/2b after six month of antiviral therapy. Genotype 3a showed significant response to therapy after three month. 32 among 374 (8.5%) were positive after 24 weeks of treatment with interferon, 29 (7.7%) patients have same genotype while 3 patients were re-infected with different HCV strains. With DAAs, only 27 (4.8%) patients were positive among 558 after 2 weeks and one patient re-infected with different genotype. Early and sustained virological response noted in DAAs. ALT and viral load decreased faster with DAAs that not achieved after 4 weeks with pegylated interferon. Sustained virological response appears in DAAs and recurrence rate is high in interferon therapy compared to DAAs. Therefore, reinfection has implications for correct treatment efficiency and to select strategies for retreatment cases.

Embora aumentem as taxas de resposta concomitantes no vírus da hepatite C (HCV), há risco de recidiva após o tratamento. Portanto, é um requisito terrível avaliar a resposta do interferon peguilado e antivirais de ação direta em Punjab, Paquistão. O estudo foi conduzido para encontrar a taxa de recorrência da infecção por HCV após o tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão. Este estudo foi conduzido no Departamento de Patologia Nawaz Sharif Medical College Gujrat, enquanto os efeitos do tratamento foram monitorados em diferentes hospitais públicos e privados de Punjab, Paquistão. Total de 973 pacientes que administraram a dose recomendada foram divididos em dois grupos: (i) Terapia baseada em interferon, (ii) antivirais de ação direta (DAAs). Outros parâmetros como ALT e carga viral foram estudados. A taxa de recorrência foi maior em mulheres infectadas com o genótipo 2b e em homens com genótipo misto 3a / 2b após seis meses de terapia antiviral. O genótipo 3a mostrou resposta significativa à terapia após três meses. 32 entre 374 (8,5%) foram positivos após 24 semanas de tratamento com interferon, 29 (7,7%) pacientes têm o mesmo genótipo, enquanto 3 pacientes foram reinfectados com diferentes cepas de HCV. Com DAAs, apenas 27 (4,8%) pacientes foram positivos entre 558 após duas semanas e um paciente reinfectado com genótipo diferente. Resposta virológica precoce e sustentada observada em DAAs. ALT e carga viral diminuíram mais rapidamente com DAAs, que não alcançou após 4 semanas com interferon peguilado. A resposta virológica sustentada aparece em DAAs, e a taxa de recorrência é alta na terapia com interferon em comparação com DAAs. Portanto, a reinfecção tem implicações para a eficiência do tratamento correto e para selecionar estratégias para casos de retratamento.

Recurrence of hepatitis C virus after treatment with pegylated interferon and direct acting antivirals in Punjab Pakistan / Recorrência do vírus da hepatite C após tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão

Although increased response rates concomitant in hepatitis C virus but relapse after treatment is threatened. Therefore, it is terrible requirement to evaluate the response of Pegylated interferon and direct acting antivirals in Punjab Pakistan. The study was conducted to find the rate of recurrence of HCV infection after treatment with Pegylated Interferon and Direct Acting Antivirals in Punjab Pakistan. This study was conducted at Department of Pathology, Nawaz Sharif Medical College Gujrat, while treatment effects monitored in different Government and Private Hospitals of Punjab, Pakistan. Total 973 patients who administered the recommended dose and divided in two groups (i) Interferon based therapy (ii) direct acting antivirals (DAAs).Other parameters like ALT and viral load studied. The rate of recurrence was higher in female infected with genotype 2b and in male with mixed genotype 3a/2b after six month of antiviral therapy. Genotype 3a showed significant response to therapy after three month. 32 among 374 (8.5%) were positive after 24 weeks of treatment with interferon, 29 (7.7%) patients have same genotype while 3 patients were re-infected with different HCV strains. With DAAs, only 27 (4.8%) patients were positive among 558 after 2 weeks and one patient re-infected with different genotype. Early and sustained virological response noted in DAAs. ALT and viral load decreased faster with DAAs that not achieved after 4 weeks with pegylated interferon. Sustained virological response appears in DAAs and recurrence rate is high in interferon therapy compared to DAAs. Therefore, reinfection has implications for correct treatment efficiency and to select strategies for retreatment cases.(AU)

Embora aumentem as taxas de resposta concomitantes no vírus da hepatite C (HCV), há risco de recidiva após o tratamento. Portanto, é um requisito terrível avaliar a resposta do interferon peguilado e antivirais de ação direta em Punjab, Paquistão. O estudo foi conduzido para encontrar a taxa de recorrência da infecção por HCV após o tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão. Este estudo foi conduzido no Departamento de Patologia Nawaz Sharif Medical College Gujrat, enquanto os efeitos do tratamento foram monitorados em diferentes hospitais públicos e privados de Punjab, Paquistão. Total de 973 pacientes que administraram a dose recomendada foram divididos em dois grupos: (i) Terapia baseada em interferon, (ii) antivirais de ação direta (DAAs). Outros parâmetros como ALT e carga viral foram estudados. A taxa de recorrência foi maior em mulheres infectadas com o genótipo 2b e em homens com genótipo misto 3a / 2b após seis meses de terapia antiviral. O genótipo 3a mostrou resposta significativa à terapia após três meses. 32 entre 374 (8,5%) foram positivos após 24 semanas de tratamento com interferon, 29 (7,7%) pacientes têm o mesmo genótipo, enquanto 3 pacientes foram reinfectados com diferentes cepas de HCV. Com DAAs, apenas 27 (4,8%) pacientes foram positivos entre 558 após duas semanas e um paciente reinfectado com genótipo diferente. Resposta virológica precoce e sustentada observada em DAAs. ALT e carga viral diminuíram mais rapidamente com DAAs, que não alcançou após 4 semanas com interferon peguilado. A resposta virológica sustentada aparece em DAAs, e a taxa de recorrência é alta na terapia com interferon em comparação com DAAs. Portanto, a reinfecção tem implicações para a eficiência do tratamento correto e para selecionar estratégias para casos de retratamento.(AU)

Genome-Wide Association Study on Two Immune-Related Traits in Jinghai Yellow Chicken

This study was designed to discover molecular marker associated with the interferon INF-γ and avian influenza (AI) antibody titer traits in Jinghai Yellow chicken (Gallus gallus). Serum samples were taken from 400 female chickens and the INF-γ concentrations and AI antibody titer levels were measured. A genome-wide association study was carried out using specific-locus amplified fragment (SLAF) sequencing. Bioinformatics analysis was applied to detect single-nucleotide polymorphisms (SNPs) associated with the two traits. After sequencing and quality control, 103,680 SLAFs and 90,961 SNPs were obtained. The 400 samples were divided into 10 subgroups to reduce the effects of group stratification. The Bonferroni adjusted P-value of genome-wide significance was set at 1.87E−06 according to the number of independent SNP markers and linkage disequilibrium blocks. A SNP that was significantly associated with INF-γ concentration was detected in the myomesin 1 (MYOM1) gene on chromosome 2, and another SNPthat was significantly associated with the AI antibody titer level was detected in an RNA methyltransferase gene (Nsun7), which was found to have an important biological function. We propose that MYOM1 and Nsun7 are valuable candidate genes that influence the disease resistance characters of chicken. However, in-depth investigations are needed to determine the essential roles of these genes in poultry disease resistance and their possible application in breeding disease resistant poultry.(AU)

Immunological indices of Giant African Land snails (Archachatina marginata) improved with fixed dose of vitamin C during acute heat stress

The study determined immunological indices of Giant African Land snail (Archachatina marginata) improved with fixed dose of vitamin C under acute heat stress (AHS). Prior to the AHS, vitamin C was administered for four weeks to two treatment groups, while other two treatment groups were not. Each treatment was monitored, haemolymph collected at 0, 30 and 60 minutes exposure times. Immunological cytokines: interferon gamma (IFN-γ) and interleukin 2 (IL-2); and total haemocyte counts (THC) were determined. Under AHS, vitamin C elevated (p <0.05) IFN-γ production (606.33 ± 302.86) compared to other groups with or without vitamin C administration (7.20 ± 1.58 vs. 73.20 ± 32.23 vs. 7.80 ± 1.36). IL-2 was not affected (p >0.05) by vitamin C under AHS. Highest (p <0.05) THC values was obtained with vitamin C administration under AHS, but reduced under no AHS. Exposure time affected (p <0.05) IFN-γ production and THC values, but not IL-2 (p >0.05). With fixed dose of vitamin C and exposure time, highest (p <0.05) IFN-γ values were obtained under AHS with vitamin C administration at 30 minutes and at 60 minutes in THC, compared to other groups. The study concluded that fixed dose of vitamin C at 150 mg kg-1 of feed was appropriate under AHS to boost the immune system of the animals.(AU)

Evaluation of interferon-stimulated gene 15 (ISG15) expression in blood neutrophils in beef cattle poisoned by Senecio spp / Avaliação da expressão do gene estimulado por interferon 15 (ISG15) em neutrófilos sanguíneos em bovinos de corte intoxicados por Senecio spp

This study aimed to assess liver damage and interferon-stimulated gene 15 (ISG15) blood expression as a consequence of embryonic signaling on maternal recognition of pregnancy in beef cattle presenting natural ingestion of Senecio spp. Epidemiological aspects, as the presence of the plant, associated to gamma glutamyl transferase (GGT) activity can be used as Senecio spp. poisoning diagnosis. Maternal recognition of pregnancy period occurs when the embryo secretes interferon tau (IFNT) to signal its presence to the mother and eventually extend corpus luteum (CL) lifespan. In our study, liver damage was determined by concentration serum GGT, cytological and histopathological examinations. Reproductive status was evaluated by concentration of progesterone, CL diameter and ISG15 mRNA expression on Day 19 following fixed-time artificial insemination (FTAI). Cows were categorized into two groups based on concentration of GGT: Group 1 (GGT<30U/L) and 2 (GGT>31U/L). No difference on body condition scores was observed. All the cows presented liver damage based on cytology and histopathological exams. Cows from the Group 1 had higher pregnancy rate, presenting larger CL diameter and greater concentration of progesterone. Interestingly, ISG15 mRNA expression had no difference between Groups 1 and 2, even presenting difference in pregnancy status. These findings suggest embryonic loss beyond Day 19. It suggests late embryonic mortality may be associated to liver insufficiency. In conclusion, liver injury and/or concentration of GGT does not alter ISG15 expression on blood neutrophils, however cows presenting lower concentration of GGT (<30U/L) had increased pregnancy status. Therefore, the concentration of GGT allow us to screen liver status and foresee a successful pregnancy in beef cattle.(AU)

O objetivo deste estudo foi avaliar a lesão hepática e a expressão sanguínea do gene estimulado por interferon 15 (ISG15) durante a sinalização embrionária, no reconhecimento materno da gestação, em bovinos de corte apresentando ingestão natural de Senecio spp. Fatores epidemiológicos, como a presença da planta, associados à atividade da gama glutamil transferase (GGT) podem ser utilizados como diagnóstico da intoxicação por Senecio spp. O reconhecimento materno da gestação ocorre quando o embrião secreta interferon tau (IFNT) para sinalizar sua presença à mãe. Em nosso estudo, a lesão hepática foi determinada pela concentração sérica de GGT, pelos exames citológicos e histopatológicos. O estado reprodutivo foi avaliado pela concentração de progesterona, diâmetro de corpo lúteo (CL) e expressão de mRNA ISG15 no Dia 19 após a inseminação artificial em tempo fixo (IATF). As vacas foram separadas em dois grupos com base na concentração de GGT sanguíneo: Grupo 1 (GGT<30U/L) e Grupo 2 (GGT>31U/L). Não foi observada nenhuma diferença no escore de condição corporal entre os grupos. Na citologia e nos exames histopatológicos todas as vacas apresentaram lesão hepática. As vacas do Grupo 1 apresentaram maior taxa de prenhez, maior diâmetro do CL e maior concentração de progesterona. Diferente do esperado, a expressão do mRNA ISG15 não foi diferente entre os Grupos 1 e 2, mesmo apresentando diferença na taxa de prenhez. Esses achados sugerem perda embrionária após o Ddia 19. Isso demonstra que a mortalidade embrionária tardia pode estar associada à insuficiência hepática. Dessa forma, conclui-se que a lesão hepática e/ou concentração de GGT não altera a expressão de ISG15 nos neutrófilos sanguíneos, porém vacas com menor concentração de GGT (<30U/L) apresentaram maiores taxas de prenhez. Assim, a concentração de GGT nos permite avaliar a saúde hepática e prever uma gestação bem-sucedida em bovinos de corte.(AU)

Evaluation of interferon-stimulated gene 15 (ISG15) expression in blood neutrophils in beef cattle poisoned by Senecio spp / Avaliação da expressão do gene estimulado por interferon 15 (ISG15) em neutrófilos sanguíneos em bovinos de corte intoxicados por Senecio spp

This study aimed to assess liver damage and interferon-stimulated gene 15 (ISG15) blood expression as a consequence of embryonic signaling on maternal recognition of pregnancy in beef cattle presenting natural ingestion of Senecio spp. Epidemiological aspects, as the presence of the plant, associated to gamma glutamyl transferase (GGT) activity can be used as Senecio spp. poisoning diagnosis. Maternal recognition of pregnancy period occurs when the embryo secretes interferon tau (IFNT) to signal its presence to the mother and eventually extend corpus luteum (CL) lifespan. In our study, liver damage was determined by concentration serum GGT, cytological and histopathological examinations. Reproductive status was evaluated by concentration of progesterone, CL diameter and ISG15 mRNA expression on Day 19 following fixed-time artificial insemination (FTAI). Cows were categorized into two groups based on concentration of GGT: Group 1 (GGT<30U/L) and 2 (GGT>31U/L). No difference on body condition scores was observed. All the cows presented liver damage based on cytology and histopathological exams. Cows from the Group 1 had higher pregnancy rate, presenting larger CL diameter and greater concentration of progesterone. Interestingly, ISG15 mRNA expression had no difference between Groups 1 and 2, even presenting difference in pregnancy status. These findings suggest embryonic loss beyond Day 19. It suggests late embryonic mortality may be associated to liver insufficiency. In conclusion, liver injury and/or concentration of GGT does not alter ISG15 expression on blood neutrophils, however cows presenting lower concentration of GGT (<30U/L) had increased pregnancy status. Therefore, the concentration of GGT allow us to screen liver status and foresee a successful pregnancy in beef cattle.(AU)

O objetivo deste estudo foi avaliar a lesão hepática e a expressão sanguínea do gene estimulado por interferon 15 (ISG15) durante a sinalização embrionária, no reconhecimento materno da gestação, em bovinos de corte apresentando ingestão natural de Senecio spp. Fatores epidemiológicos, como a presença da planta, associados à atividade da gama glutamil transferase (GGT) podem ser utilizados como diagnóstico da intoxicação por Senecio spp. O reconhecimento materno da gestação ocorre quando o embrião secreta interferon tau (IFNT) para sinalizar sua presença à mãe. Em nosso estudo, a lesão hepática foi determinada pela concentração sérica de GGT, pelos exames citológicos e histopatológicos. O estado reprodutivo foi avaliado pela concentração de progesterona, diâmetro de corpo lúteo (CL) e expressão de mRNA ISG15 no Dia 19 após a inseminação artificial em tempo fixo (IATF). As vacas foram separadas em dois grupos com base na concentração de GGT sanguíneo: Grupo 1 (GGT<30U/L) e Grupo 2 (GGT>31U/L). Não foi observada nenhuma diferença no escore de condição corporal entre os grupos. Na citologia e nos exames histopatológicos todas as vacas apresentaram lesão hepática. As vacas do Grupo 1 apresentaram maior taxa de prenhez, maior diâmetro do CL e maior concentração de progesterona. Diferente do esperado, a expressão do mRNA ISG15 não foi diferente entre os Grupos 1 e 2, mesmo apresentando diferença na taxa de prenhez. Esses achados sugerem perda embrionária após o Ddia 19. Isso demonstra que a mortalidade embrionária tardia pode estar associada à insuficiência hepática. Dessa forma, conclui-se que a lesão hepática e/ou concentração de GGT não altera a expressão de ISG15 nos neutrófilos sanguíneos, porém vacas com menor concentração de GGT (<30U/L) apresentaram maiores taxas de prenhez. Assim, a concentração de GGT nos permite avaliar a saúde hepática e prever uma gestação bem-sucedida em bovinos de corte.(AU)

Possible impact of neutrophils on immune responses during early pregnancy in ruminants

The interaction between early embryo and maternal immune system for the establishment of pregnancy is the focus of several studies; however, it remains unclear. The maternal immune response needs to keep a balance between avoiding any damage to the conceptus and maintaining its function in combating microbes as well. When conceptus-maternal crosstalk cannot achieve this balance, pregnancy losses might occur. Intercommunication between mother and conceptus is fundamental during early pregnancy to dictate the outcome of pregnancy. In ruminants, the embryo reacts with the maternal system mainly via interferon tau (IFNT) release. IFNT can act locally on the embryo and endometrial cells and systemically in several tissues and cells to regulate their response via the expression of interferon-stimulated genes (ISGs). Also, IFNT can induce the expression of inflammatory-related genes in immune cells. Day 7 embryo induces a shift in the maternal immune response towards anti-inflammatory (Th2) immune responses. During maternal recognition of pregnancy, peripheral mononuclear cells (PBMCs) and polymorphonuclear cells (PMNs) express markers that configure an anti-inflammatory response. However, PMNs response is more sensitive to the effects of IFNT. PMNs are more likely to express interferon-stimulated genes (ISGs), transforming growth factor-beta (TGFB), interleukin 10 (IL10), and arginase-1 (ARG1), configuring one of the most rapid immune responses to early pregnancy. This review focus on the local and peripheral immune responses during early pregnancy in ruminants, mainly the PMNs function in the immune system.(AU)

Actions of interferon tau during maternal recognition of pregnancy / Ações do interferon tau durante o reconhecimento materno da gestação

Maternal recognition of pregnancy in ruminants is a physiological process that requires an interaction between the conceptus and the mother in order to avoid luteal regression. Studies on 60s started to investigate the communication, and late on the 80s it was identified a Type I interferon called interferon tau. In the 90s the local action of interferon tau was described. It suppresses luteolytic pulses of prostaglandin F2 alpha inhibiting endometrial expression of estrogen and oxytocin receptors. Several studies presented the expression of interferonstimulated genes on extrauterine tissues. After, the endocrine action of interferon tau was described. This study revealed a greater interferon bioactivity in the serum of uterine vein from pregnant ewes when compared to nonpregnant ewes. Following a series of experiments were preformed to better understand the endocrine action of interferon tau in several extrauteine tissues. This review presents the endocrine action of interferon tau during maternal recognition of pregnancy period in ruminants.(AU)

O reconhecimento materno da gestação em ruminantes é um processo fisiológico que requer a interação entre o concepto e a mãe com o obejetivo de evitar a regressão luteal. Estudos na década de 1960 começaram a investigar essa comunicação entre concepto e mãe, e durante a década de 1980 foi identificado um tipo I de interferon que foi denominado interferon tau. Nos anos 1990, foi descrita a ação local do interferon tau. Essa ação suprime os pulsos luteolíticos de prostaglandina F2 alfa inibindo a expressão endometrial dos receptors de estrógeno e ocitocina. Muitos estudos têm apresentado a expressão dos genes estimulados por interferons nos tecidos extrauterinos. Posteriormente, a ação endócrina do interferon tau também foi descrita. Este estudo revelou uma maior bioatividade de interferon no soro da veia uterina de ovelhas prenhes quando comparada com ovelhas não gestantes. Após, uma série de experimentos foram realizados para melhor compreender a ação endócrina do interferon tau em vários tecidos extrauterinos. Esta revisão destaca a ação endócrina do interferon tau durante o período de reconhecimento materno da gestação em ruminantes.(AU)

Actions of interferon tau during maternal recognition of pregnancy / Ações do interferon tau durante o reconhecimento materno da gestação

Maternal recognition of pregnancy in ruminants is a physiological process that requires an interaction between the conceptus and the mother in order to avoid luteal regression. Studies on 60’s started to investigate the communication, and late on the 80’s it was identified a Type I interferon called interferon tau. In the 90’s the local action of interferon tau was described. It suppresses luteolytic pulses of prostaglandin F2 alpha inhibiting endometrial expression of estrogen and oxytocin receptors. Several studies presented the expression of interferonstimulated genes on extrauterine tissues. After, the endocrine action of interferon tau was described. This study revealed a greater interferon bioactivity in the serum of uterine vein from pregnant ewes when compared to nonpregnant ewes. Following a series of experiments were preformed to better understand the endocrine action of interferon tau in several extrauteine tissues. This review presents the endocrine action of interferon tau during maternal recognition of pregnancy period in ruminants.

O reconhecimento materno da gestação em ruminantes é um processo fisiológico que requer a interação entre o concepto e a mãe com o obejetivo de evitar a regressão luteal. Estudos na década de 1960 começaram a investigar essa comunicação entre concepto e mãe, e durante a década de 1980 foi identificado um tipo I de interferon que foi denominado interferon tau. Nos anos 1990, foi descrita a ação local do interferon tau. Essa ação suprime os pulsos luteolíticos de prostaglandina F2 alfa inibindo a expressão endometrial dos receptors de estrógeno e ocitocina. Muitos estudos têm apresentado a expressão dos genes estimulados por interferons nos tecidos extrauterinos. Posteriormente, a ação endócrina do interferon tau também foi descrita. Este estudo revelou uma maior bioatividade de interferon no soro da veia uterina de ovelhas prenhes quando comparada com ovelhas não gestantes. Após, uma série de experimentos foram realizados para melhor compreender a ação endócrina do interferon tau em vários tecidos extrauterinos. Esta revisão destaca a ação endócrina do interferon tau durante o período de reconhecimento materno da gestação em ruminantes.

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1, HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.

Parthenogenetic bovine embryos secrete type I interferon capable of stimulating ISG15 in luteal cell culture

Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1, HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.(AU)

Benznidazole affects expression of Th1, Th17 and Treg cytokines during acute experimental Trypanosoma cruzi infection

Background The present study evaluated the effect of treatment with benznidazole on mRNA expression of IFN-γ, IL-17, IL-10, TGF-β and FoxP3 in spleen and heart tissue of BALB/c mice in the acute phase of an experimental infection with Trypanosoma cruzi, strains JLP or Y. Methods The mRNA expression of cytokines and parasite load were assessed by q-PCR. Dependent groups were compared using Student's paired t-test and independent groups were compared using Student's unpaired t-test. Results Infection with the JLP or Y strains increased expression of IFN-γ in the heart and of IL-10 and IL-17 in the spleen and heart compared to uninfected animals. Treatment increased the expression of IFN-γ and decreased the expression of IL-17, IL-10, TGF- β and Foxp3 in spleen and heart tissue compared to untreated infected animals. Conclusion Benznidazole can induce Th1 profile in the initial of the acute phase. The treatment decreased the parasite load in both organs, although the number of parasites in Y-strain-infected mice remained high. The data suggest that benznidazole may modulate cytokine expression in infection and can be dependent of the strain. However, treatment was not fully effective in the infection provoked by Y strain, probably due to the characteristics of the strain itself.(AU)

Benznidazole affects expression of Th1, Th17 and Treg cytokines during acute experimental Trypanosoma cruzi infection

Background The present study evaluated the effect of treatment with benznidazole on mRNA expression of IFN-γ, IL-17, IL-10, TGF-β and FoxP3 in spleen and heart tissue of BALB/c mice in the acute phase of an experimental infection with Trypanosoma cruzi, strains JLP or Y. Methods The mRNA expression of cytokines and parasite load were assessed by q-PCR. Dependent groups were compared using Student's paired t-test and independent groups were compared using Student's unpaired t-test. Results Infection with the JLP or Y strains increased expression of IFN-γ in the heart and of IL-10 and IL-17 in the spleen and heart compared to uninfected animals. Treatment increased the expression of IFN-γ and decreased the expression of IL-17, IL-10, TGF- β and Foxp3 in spleen and heart tissue compared to untreated infected animals. Conclusion Benznidazole can induce Th1 profile in the initial of the acute phase. The treatment decreased the parasite load in both organs, although the number of parasites in Y-strain-infected mice remained high. The data suggest that benznidazole may modulate cytokine expression in infection and can be dependent of the strain. However, treatment was not fully effective in the infection provoked by Y strain, probably due to the characteristics of the strain itself.(AU)

Effects of thymoquinone on interleukin-1 and interferon gamma gene expression and antibody titers against newcastle disease in broiler chickens under oxidative stress

An experiment was conducted to determine the effects of the dietary inclusion of different levels of thymoquinone (TQ) of broilers subjected to oxidative stress or not on the antibody titers against Newcastle disease and on the gene expression of interleukine-1 and interferon gamma. A total of 320 one-day-old broilers was randomly assigned to eight treatments with four replicates of 10 birds each, in a 4 × 2 factorial arrangement, consisting of four thymoquinone (TQ) levels (0, 5, 8, or 11 mg/kg body weight) and two levels tert-butyl hydroperoxide (t-BHP) injection (0 or 0.02 mmol/kg of body weight). Blood samples were collected from two birds per replicate to determine antibody titers against Newcastle disease. At the end of experiment, two birds per replicate were randomly selected, sacrificed and their spleens were collected to evaluate the genes expressioninterleukin-1 and interferon gamma (p 0.05). The dietary inclusion of TQ of broilers subjected or not oxidative stress increased antibody production against Newcastle disease (p 0.05). Both individual and combined dietary inclusion of t-BHP and TQ promote the differentiation and proliferation of spleen cells and the gene expression of interleukin-1 and interferon gamma (p 0.05).

Effects of thymoquinone on interleukin-1 and interferon gamma gene expression and antibody titers against newcastle disease in broiler chickens under oxidative stress

An experiment was conducted to determine the effects of the dietary inclusion of different levels of thymoquinone (TQ) of broilers subjected to oxidative stress or not on the antibody titers against Newcastle disease and on the gene expression of interleukine-1 and interferon gamma. A total of 320 one-day-old broilers was randomly assigned to eight treatments with four replicates of 10 birds each, in a 4 × 2 factorial arrangement, consisting of four thymoquinone (TQ) levels (0, 5, 8, or 11 mg/kg body weight) and two levels tert-butyl hydroperoxide (t-BHP) injection (0 or 0.02 mmol/kg of body weight). Blood samples were collected from two birds per replicate to determine antibody titers against Newcastle disease. At the end of experiment, two birds per replicate were randomly selected, sacrificed and their spleens were collected to evaluate the genes expressioninterleukin-1 and interferon gamma (p 0.05). The dietary inclusion of TQ of broilers subjected or not oxidative stress increased antibody production against Newcastle disease (p 0.05). Both individual and combined dietary inclusion of t-BHP and TQ promote the differentiation and proliferation of spleen cells and the gene expression of interleukin-1 and interferon gamma (p 0.05). (AU)

Advances in reproductive management: pregnancy diagnosis in ruminants

During the last 10 years the U.S. dairy industry has experienced a reversal of the decades-long trend in declining fertility traits. In fact, there is evidence that, nationally, this is contributing to improvements in pregnancy rates. And while these measures are still close to their historical lows, there is reason for optimism that this reversal will continue into the future. The reasons for improved pregnancy rates are related to use of biotechnolog ies and improved management practices for high producing dairy cows as well as greater emphasis on genetic selection for fertility - related traits. Combined, these factors have resulted in a reduction in the average days to first service in our national dairy herd of approximately 10 day s over the past decade and a reduction in calving interval of approximately 15 days. However, current challenges include accurate identification of cows that fail to conceive following insemination and their timely reinsemination. The primary metric for success of pregnancy diagnosis is the inter-service interval, or the number of days between insemination and the subsequent insemination in a cow that fails to conceive or that loses an established pregnancy. This trait is directly affected by the choice of pregnancy diagnosis method. Pregnancy diagnosis methods include estrous detection (visual or assisted), transrectal palpation of uterine contents, transrectal ultrasound visualization of uterine contents and assay for hormones in blood, milk or other body fluids. Each of these methods has advantages and disadvantages. Presently, ultrasound and blood hormone assay at 28 days after insemination offer the earliest specific diagnostics for determining pregnancy status. However, other methods are on the horizon that may provide opportunities to further reduce the interval between insemination and accurate diagnosis of pregnancy status of dairy cattle [...]

Advances in reproductive management: pregnancy diagnosis in ruminants

During the last 10 years the U.S. dairy industry has experienced a reversal of the decades-long trend in declining fertility traits. In fact, there is evidence that, nationally, this is contributing to improvements in pregnancy rates. And while these measures are still close to their historical lows, there is reason for optimism that this reversal will continue into the future. The reasons for improved pregnancy rates are related to use of biotechnolog ies and improved management practices for high producing dairy cows as well as greater emphasis on genetic selection for fertility - related traits. Combined, these factors have resulted in a reduction in the average days to first service in our national dairy herd of approximately 10 day s over the past decade and a reduction in calving interval of approximately 15 days. However, current challenges include accurate identification of cows that fail to conceive following insemination and their timely reinsemination. The primary metric for success of pregnancy diagnosis is the inter-service interval, or the number of days between insemination and the subsequent insemination in a cow that fails to conceive or that loses an established pregnancy. This trait is directly affected by the choice of pregnancy diagnosis method. Pregnancy diagnosis methods include estrous detection (visual or assisted), transrectal palpation of uterine contents, transrectal ultrasound visualization of uterine contents and assay for hormones in blood, milk or other body fluids. Each of these methods has advantages and disadvantages. Presently, ultrasound and blood hormone assay at 28 days after insemination offer the earliest specific diagnostics for determining pregnancy status. However, other methods are on the horizon that may provide opportunities to further reduce the interval between insemination and accurate diagnosis of pregnancy status of dairy cattle [...](AU)

Cellular immune response of Curraleiro Pé-duro and Nellore calves following Mycobacterium bovis-BCG vaccination / Resposta imune celular de bezerros Curraleiro Pé-duro e Nelore após vacinação com Mycobacterium bovis-BCG

The present study aimed to assess the CD4, CD8 and Gamma delta T cells blood levels for Curraleiro Pé-duro, as well as the specific IFN-γ response after BCG vaccination using flow cytometry. The specific immune response against BCG was also evaluated by tuberculin skin test, performed before and 45 days after the vaccination. For comparison purposes, the same parameters were investigated on Nellore calves, an exotic bovine with resistance previously demonstrated. Naturally, Curraleiro Pé-duro animals had greater levels of CD4, CD8 and Gamma delta lymphocytes (p<0.05). In response to vaccine, Curraleiro Pé-duro showed greater ability to respond specifically to BCG, generating resistance profile (Th1), evidenced by greater number of antigen specific CD4+ cells producing IFN-γ (p<0.05) and also higher tuberculin skin test reaction (p<0.05). Additionally, vaccinated Curraleiro Pé-duro calves had higher CD4 cells numbers than both Nellore control (p<0.05) and vaccinated groups (p<0.05). Curraleiro Pé-duro calves' higher basal lymphocytes blood level and stronger response in both IFN-γ and tuberculin skin test parameters probably play a positive role on protection/resistance to Mycobacterium bovis. (AU)

O presente estudo teve como objetivo avaliar níveis sanguíneos de células CD4, CD8 e células T gama-delta no sangue periférico de bezerros Curraleiro Pé-Duro, bem como a produção específica de IFN-γ por essas células em resposta à vacinação com BCG, através de citometria de fluxo. A resposta imune específica contra BCG também foi avaliada por teste tuberculínico, realizado antes e 45 dias após a vacinação. Para fins de comparação, os mesmos parâmetros foram investigados em bezerros da raça Nelore, uma raça bovina exótica com resistência demonstrado anteriormente. Naturalmente, animais da raça Curraleiro Pé-Duro apresentaram maiores níveis de CD4, CD8 e linfócitos gama-delta. Em resposta a vacina, Curraleiro Pé-duro mostrou maior capacidade de responder especificamente ao BCG, gerando perfil de resistência (Th1), evidenciado pelo maior número de células CD4+ específicas produtoras de IFN-γ e maior reação cutânea a por tuberculina. Os maiores níveis basais de linfócitos, maior produção de IFN-γ e reação cutânea à prova tuberculínica provavelmente desempenham um papel positivo na proteção/resistência ao Mycobacterium tuberculosis. (AU)

Evaluation of prognostic value of cell adhesion molecules in chronic hepatitis C therapy

The most reliable determination of severity and prognosis in chronic viral hepatitis is the histological staging of the disease, which comprises an invasive procedure and is often not well accepted by patients. The search for alternative non-invasive methods is mandatory especially in follow-ups after initial assessment by biopsy. The aim of this study was to evaluate the role of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in patients under interferon alpha therapy whether responsive or non-responsive to therapy. Thirty chronic hepatitis C patients (CHC) under combined therapy of interferon-α with ribavirin, whether responsive or non-responsive, were included in the study as well as ten healthy controls. Serum VCAM-1 and ICAM-1 levels were calculated using commercial enzyme linked immunosorbent assay (ELISA) kits. Before the therapy, sICAM-1 and sVCAM-1 levels were significantly higher among CHC patients (96.9 ± 39.74 and 679.4 ± 218.98 ng/mL, respectively) than in the control group (14.3 ± 4.32 and 108.9 ± 49.21 ng/mL, respectively), (p < 0.05 for both). They were higher among non-responsive than in responsive patients. Comparisons in soluble ICAM-1 (sICAM-1) levels of responsive persons during treatment revealed a statistically significant increase at baseline (81.27 ± 25.797) versus its value after one month (52.33 ± 12.76), p < 0.05 and after three months (49.67 ± 14.42), p < 0.05. However, no statistically significant difference was detected between one and three-month sICAM-1 levels post-therapy commencement (p = 0.055). Also, no statistically significant difference was detected between sVCAM-1 levels at baseline (521.47 ± 137.29) versus three months after therapy initiation (517.53 ± 130.6), p = 0.854. The occurrence of a significant decrease in sICAM-1 level as early as one month after therapy in responsive patients only allows us to conclude that sICAM-1 could be used as an early marker in CHC patients under interferon therapy to predict prognosis and guide the treatment, whereas sVCAM-1 does not present any difference between the studied groups.(AU)

Quantificação do interferon-tau durante o reconhecimento materno da gestação em fêmeas bovinas Bos taurus indicus / Quantification of interferon-tau during the maternal recognition of pregnancy in Bos taurus indicus cows

Durante o período crítico do reconhecimento materno, compreendidoentre o 15º e 19º dias da gestação, o concepto deve sintetizarcompetentemente moléculas capazes de bloquear a síntese deprostaglandina F2α (PGF2α) e a luteólise. Em bovinos, a principalmacromolécula protéica envolvida em tal bloqueio é o interferon-tau(IFN-τ). Durante o período crítico, falhas neste reconhecimentodeterminam à mortalidade embrionária em até 40% das fêmeasinseminadas. Informações sobre o IFN-τ em animais Bos taurus indicus,ainda são restritas. Este estudo objetivou uma avaliação quantitativado IFN-τ durante o período crítico do reconhecimento materno, emlavados uterinos obtidos por sonda de Foley (dias 14, 16 e 18 pósestro) ou post-mortem (dia 18 pós-estro). Para tanto, foram utilizadasfêmeas multíparas azebuadas (Bos taurus indicus), cíclicas ou prenhes,nos dias 14, 16 e 18 pós-estro. Para a obtenção dos lavados, os úterosforam infundidos com solução de Ringer Simples. Os lavados foramconcentrados por ultra-filtração e liofilizados. As macromoléculasprotéicas foram separadas por Eletroforese Unidimensional SDSPAGE, em gel com 15% de poliacrilamida. A quantificação doIFN-τ nos lavados uterinos foi realizada por Western-Blotting edensitometria. Tanto nos lavados obtidos por sonda de Foley quantonos post-mortem foi possível observar bandas de proteínas queapresentaram reação cruzada com os anticorpos utilizados no WesternBlotting. O IFN-τ foi detectado apenas nos lavados uterinospost-mortem de vacas prenhes (P<0,05). A densidade óptica não foiafetada pelo dia do período crítico, estado (cíclico ou prenhe) ouinteração dia x estado. Nos lavados post-mortem não houve efeito depeso do concepto ou concentração de progesterona plasmática no diado lavado na densidade da banda protéica referente ao IFN-τ .(AU)

During the critical period of the maternal recognition, which occursbetween days 15 and 19 of pregnancy, the conceptus must competentlysynthesize molecules capable of blocking the synthesis ofprostaglandin F2α (PGF2α) and luteolysis. In cattle, the majormacromolecule involved in suck blockage is the protein interferontau (IFN-τ). During the critical period, failures in the recognition ofpregnancy determine embryonic mortality on up to 40% ofinseminated cows. Data about IFN-τ in Bos taurus indicus are stillscarce. Objective of this study was to quantitatively evaluate the presenceof IFN-τ during the critical period for maternal recognition ofpregnancy in uterine flushings obtained in vivo by Foley catheter (Days14, 16 and 18 post estrus) or post-mortem (Day 18 post estrus).Multiparous, cyclic or pregnant zebu cows (Bos taurus indicus) on days14, 16 and 18 post estrus were used for in vivo or post mortem uterineflushing collection. In both cases, a Ringer solution was used to washthe uterus of cows. Uterine flushings were concentrated by ultrafiltration and lyophilized. Proteins were separated by one-dimensionalelectrophoresis (SDS-PAGE) in a 15% polyacrilamide gel. Interferontau quantification in uterine flushings was performed by westernblotting and densitometry. Non-specific protein bands were observedin both in vivo and post mortem uterine flushings. Interferon-tau wasdetected only in uterine flushings obtained from pregnant cowspost-mortem (P<0.05). Optical density of protein bands was not affectedby the day of the critical period, state (cyclic or pregnant) or interactionday x state. There was no effect of the conceptus weight orprogesterone concentration on the day of uterine flushing collectionin the optical density of the IFN-τ protein band.(AU)