Use of ammonium chloride to prevent urolithiasis in sheep / [Utilização do cloreto de amônio na prevenção de urolitíase em ovinos]

Urolithiasis has a high incidence among confined sheep. It is multifactorial and may cause economic damage. Our aim was to determine the capacity of urinary acidification using ammonium chloride in sheep. Twenty-five 3-month-old male sheep were confined and randomly divided into three groups; the G200 and G500 groups received 200mg/kg/GW and 500mg/kg/GW of ammonium chloride daily for 56 consecutive days, respectively, whereas the CG group did not receive ammonium chloride. Sampling times and clinical evaluation were performed weekly, starting from the 14th day of confinement (M1 or immediately before administering ammonium chloride) until the 17th day (M9) of the feedlot. Hemogasometry, biochemical examination of serum urea and creatinine concentration and ultrasound evaluation of the urinary tract were performed. The urinalysis indicated a higher incidence of ammonium magnesium phosphate crystals at the beginning of the study, showing a migration to urate crystal formation, mainly in the G500 group because of urinary acidification. No hemogasometric, serum biochemistry, ruminal fluid, or ultrasonographic changes were observed. Urinary acidification was achieved and maintained after M7 during the administration of ammonium chloride in the G500 group, but not in the other study groups.(AU)

A urolitíase apresenta alta incidência em ovinos confinados, etiologia multifatorial, e pode causar prejuízo econômico. O objetivo do presente estudo foi determinar a capacidade da acidificação urinária mediante o uso de cloreto de amônio em ovinos. Foram utilizados 25 ovinos de três meses de idade, confinados e divididos aleatoriamente em três grupos: grupo CG (controle) não recebeu cloreto de amônio; grupo G200 (200mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos; grupo G500 (500mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos, administrados diariamente por via oral. Os momentos (M) de colheita de amostras e de avaliação clínica foram realizados com intervalo de sete dias, sendo M1 (imediatamente antes do cloreto de amônio), M2 (sete dias após) até M9, totalizando 70 dias de confinamento. Foram realizadas hemogasometria, concentração sérica de ureia e creatinina e avaliação ultrassonográfica do trato urinário. Na urinálise, houve uma maior incidência de cristais de fosfato amônio magnesiano no início do estudo, com migração para formação de cristais de urato, principalmente no G500, devido à acidificação urinária. Não houve alterações hemogasométricas, na bioquímica sérica, no líquido ruminal, ou alterações ultrassonográficas. A acidificação urinária foi obtida e mantida a partir do M7 durante a administração do cloreto de amônio no grupo G500, não ocorrendo nos outros grupos de estudo.(AU)

Prepartum anionic diet induces hyperchloremic acidosis in high-producing dairy cows without preventing subclinical hypocalcemia / Dieta aniônica pré-parto provoca acidose hiperclorêmica em vacas leiteiras de alta produção mas não evita a hipocalcemia subclínica

In this study we evaluated the effects of the prepartum anionic diet on the electrolyte balance and calcemia of high producing dairy cows in the first days of lactation, and investigated the impact on the frequency of subclinical hypocalcemia (SCH). Sixty healthy Holstein cows, producing 30 kg of milk/day, handled in intensive system (compost barn), were distributed in groups (n=15) according to lactation order: first, second, third, and fourth to sixth. In the last three weeks before calving they received a diet with negative DCAD (-6mEq/100g DM) and high chloride content. After calving, they received a diet with positive DCAD (18mEq/100g DM). Urine pH was measured before calving. Serum Na+, Cl-, K+, and total Ca concentrations, and the strong ion difference (SID3) were determined in samples taken soon after calving (0h), 24, 48, 72 and 96h after. The frequencies of SCH were determined considering the critical value of 2.125mmol/L (8.5mg/dL). Two-way repeated measures ANOVA and chi-square test were used for comparisons. The cows eliminated acidic urine before calving. Na+, K+, Cl-, and SID3 values did not differ between groups. Na+ and K+ did not vary between days; Cl- was elevated at calving and decreased until 72h; and SID3 was reduced at calving and increased up to 48h. The Ca levels were reduced until 24h and increased up to 72h. Cows of third and fourth to sixth lactations presented lower values up to 24h. SCH was observed in almost half of the cows (43.3% to 55%) until 48h. The maintenance of hypocalcemia for three or more consecutive days occurred in 53.3% of third and fourth to sixth lactations cows. Ingestion of a high chloride prepartum anionic diet led to hyperchloremic acidosis and this imbalance was reversed on the second postpartum day. The induced effects on electrolyte and acid-base balances were not able to prevent the occurrence of SCH in the first days of lactation.(AU)

Os objetivos do estudo foram avaliar os efeitos que a dieta aniônica pré-parto provoca sobre o equilíbrio eletrolítico e sobre a calcemia de vacas leiteiras de alta produção nos primeiros dias de lactação, e verificar o impacto sobre a frequência da hipocalcemia subclínica (HSC). Sessenta fêmeas hígidas HPB, com produção de 30 kg de leite/dia, manejadas em sistema intensivo (compost barn), foram distribuídas por grupos (n=15) de acordo com a ordem de lactação: primeira, segunda, terceira e quarta a sexta. Nas três semanas pré-parto receberam dieta com DCAD negativa (-6mEq/100g MS) e teor de cloreto elevado. Após o parto receberam dieta com DCAD positiva (18mEq/100g MS). O pH da urina foi mensurado antes do parto. As concentrações séricas de Na+, Cl-, K+ e Ca total e a diferença de íons fortes (SID3) foram determinadas em amostras colhidas ao parto (0h), 24, 48, 72 e 96h após. As frequências de HSC foram determinadas considerando-se o valor crítico de 2,125mmol/L (8,5mg/dL). ANOVA de medidas repetidas e teste de qui-quadrado foram empregados para as comparações. As vacas eliminavam urina ácida antes do parto. Os valores de Na+, K+, Cl- e SID3 não diferiram entre os grupos. Na+ e K+ não variaram entre os dias; Cl- era elevado ao parto e diminuiu até 72h; e SID3 era reduzida ao parto e aumentou até 48h. A calcemia era reduzida até 24h e se elevou até 72h. Vacas de terceira e de quarta a sexta lactações apresentaram valores mais baixos até 24h. A HSC foi observada em quase metade das vacas (43,3% a 55%) até 48h. A manutenção de hipocalcemia por três ou mais dias seguidos ocorreu em 53,3% das vacas de terceira e de quarta a sexta lactações. A ingestão de dieta aniônica pré-parto com alto teor de cloreto provocou acidose hiperclorêmica e este desequilíbrio se reverteu no segundo dia pós-parto. Os efeitos induzidos sobre os equilíbrios eletrolítico e ácido base não foram capazes de prevenir a ocorrência de HSC nos primeiros dias da lactação.(AU)

Alteraciones leucocitarias por exposición a perclorato de amonio en ratones de la cepa ICR / Leucocitary changes by ammonium perchlorate in ICR mice strain / Alterações leucocitárias por exposição a perclorato de amônio em camundongos da linhagem ICR

El presente trabajo tuvo como objetivo identificar alteraciones en el recuento de leucocitos en sangre periférica, generadas por la exposición a perclorato de amonio, en ratones de la cepa ICR, evaluando diferentes concentraciones de exposición y diferenciando dichas alteraciones en machos y hembras. Se realizó mediante un trabajo de diseño analítico tipo experimental. Se utilizaron 60 ratones de la cepa ICR, 30 machos y 30 hembras de los cuales 50 fueron expuestos a perclorato de amonio. La manipulación se llevó a cabo de acuerdo con lo establecido en la resolución 008430 de 1993, artículo 87 del Ministerio de Salud de la República de Colombia. Un 70% de los ratones hembra y un 83% de los ratones macho expuestos a perclorato de amonio presentaron alteraciones en el recuento celular de la línea blanca. Los machos presentaron mayor alteración leucocitaria, en especial linfocitopenia. Por otro lado, el peso en los ratones expuestos disminuyó considerablemente hacia la tercera semana de administración de perclorato de amonio, lo cual podría indicar que esta sustancia tóxica genera un estado de inmunosupresión. Se evidenció leucopenia, específicamente neutropenia, neutrofilia y linfocitopenia como principales alteraciones en el recuento de leucocitos en ratones de la cepa ICR, expuestos a perclorato de amonio.

The objective of this work was to identify the changes in the leukocyte counts in peripheral blood by ammonium perchlorate in ICR strain mice, evaluating the different exposure concentrations and differentiating the changes between males and females. Through an analytic experimental investigation, 60 ICR strain mice, 30 males and 30 females were used, 50 of which were exposed to ammonium perchlorate. This study was carried out by Resolution 008430/93, article 87 of Ministry of Health of Colombia. A total of 70% females and 83% males had changes in the leukocyte counts, especially lymphocytopenia in males, further, weight decreased the third week of treatment, and probably that toxic substance induces a state of immunosuppression. The main changes in the leukocyte counts in peripheral blood by ammonium perchlorate in ICR strain was leukopenia.

O objetivo deste trabalho foi identificar as alterações na contagem de leucócitos em sangue periférico, geradas pela exposição a perclorato de amônio em camundongos da linhagem ICR, avaliando diferentes concentrações de exposição e diferenciando estas alterações entre machos e fêmeas. Isto foi realizado através de um trabalho de desenho analítico tipo experimental. Foram utilizados 60 camundongos da linhagem ICR, sendo 30 machos e 30 fêmeas, dos quais 50 foram expostos ao perclorato de amônio. O manuseio foi realizado conforme a resolução 008430 de 1993, artigo 87 do Ministério da Saúde da Colômbia. 70% dos camundongos fêmea e 83% dos machos apresentaram alterações na contagem celular dos leucócitos, especialmente a linfocitopenia em machos. Além disso, o peso nos camundongos expostos diminuiu en forma considerável para a terceira semana de tratamento com perclorato de amônio, o qual poderia indicar que esta substância tóxica gera um estado de imunossupressão. Houve evidência de leucopenia, especificamente neutropenia, neutrofilia e linfocitopenia como principais alterações na contagem de leucócitos em camundongos da linhagem ICR, expostos a perclorato de amônio.

Effects of the Helicobacter pylori Virulence Factor CagA and Ammonium Ion on Mucins in AGS Cells

PURPOSE: To investigate the effects of Helicobacter pylori (H. pylori)-CagA and the urease metabolite NH₄⁺ on mucin expression in AGS cells. MATERIALS AND METHODS: AGS cells were transfected with CagA and/or treated with different concentrations of NH₄⁺CL. Mucin gene and protein expression was assessed by qPCR and immunofluorescence assays, respectively. RESULTS: CagA significantly upregulated MUC5AC, MUC2, and MUC5B expression in AGS cells, but did not affect E-cadherin and MUC6 expression. MUC5AC, MUC6, and MUC2 expression in AGS cells increased with increasing NH₄⁺ concentrations until reaching a peak level at 15 mM. MUC5B mRNA expression in AGS cells (NH₄⁺ concentration of 15 mM) was significantly higher than that at 0, 5, and 10 mM NH₄⁺. No changes in E-cadherin expression in AGS cells treated with NH₄⁺ were noted, except at 20 mM. The expression of MUC5AC, MUC2, and MUC6 mRNA in CagA-transfected AGS cells at an NH₄⁺ concentration of 15 mM was significantly NH₄⁺ concentration, and was significantly higher compared to that in untreated cells. No significant change in the expression of E-cadherin mRNA in CagA-transfected AGS cells was observed. Immunofluorescence assays confirmed the observed changes. CONCLUSION: H. pylori may affect the expression of MUC5AC, MUC2, MUC5B, and MUC6 in AGS cells via CagA and/or NH₄⁺, but not E-cadherin.

Induced metabolic acidosis by ammonium chloride: action mechanisms, dose and effects on athletic performance / Acidose metabólica induzida por cloreto de amônio: mecanismos de ação, dose utilizada e efeitos sobre o desempenho esportivo

ABSTRACT The relationship between metabolic acidosis and athletic performance has been investigated over the years through manipulation of the blood and muscle pH. Among the pH manipulation manners, the ammonium chloride (NH4Cl) is the most widely used chemical component when is intentioned to induce a blood acidosis status prior to exercise. However, there is a lack of studies investigating the action of this substance on athletic performance as only two studies were performed in the last 15 years. Thus, it will be addressed in the present review the main aspects involved in NH4Cl ingestion, giving a focus to the action mechanisms of this substance, specifications about the used dose and their effects on athletic performance.

RESUMO A relação entre a acidose metabólica e o desempenho esportivo tem sido investigada ao longo dos anos através de manipulações do pH sanguíneo e muscular. Entre as formas de manipulação do pH, o cloreto de amônio (NH4Cl) é o componente químico mais utilizado quando se pretende induzir um estado de acidose sanguínea previamente ao exercício. Entretanto, investigações acerca da ação desse agente sobre o desempenho esportivo ainda podem ser consideradas escassas, quando foram realizados apenas dois estudos nos últimos 15 anos. Dessa forma, serão abordados na presente revisão os principais aspectos envolvidos na ingestão de NH4Cl, dando um enfoque aos mecanismos de ação dessa substância, especificações acerca do tipo de dose utilizada e seus efeitos sobre o desempenho esportivo.

Clinical role of dietary thiamine on regulation of renal response to metabolic acidosis in adult rats / Papel clínico de tiamina na dieta sobre a regulação da resposta renal de acidose metabólica em ratos adultos

Kidney plays a central role in maintaining the composition of body fluids by regulating water, NaCl, acid base, and solute reabsorption and excretion, respectively. The study was done to investigate the physiological role of thiamine in regulation of renal response to metabolic acidosis induced by NH4Cl in adult male rats. For this experiment, fifty rats were used. They were divided into five groups. Control rats received basal diet; rats fed on basal diet mixed with NH4Cl (4g NH4Cl/100g diet) to induce severe metabolic acidosis, rats fed on basal supplemented diet with thiamine (600 mg/kg diet), and rats fed on basal supplemented diet with thiamine before and after induction of metabolic acidosis by NH4Cl for 14 days. The results showed that the plasma levels of chloride, urea, and creatinine were significantly elevated in metabolic acidosis induced by NH4Cl. Thiamine supplementation at high dose before or after induction improved the chloride values. Feeding diets supplemented with thiamine modulated the plasma sodium and bicarbonate values. Supplementation with vitamin B1 as preventive agent significantly restored these changes to near control value and when used as curative agent improved plasma creatinine and urea levels. Urinary pH and potassium levels were decreased significantly in metabolic acidotic rats when compared to all experimental groups. Urinary ammonia and aldosterone levels were decreased by thiamine supplementation as protective agent. Supplementation with vitamin B1 as preventive and curative agents, restored the affected parameters and regulate the response of kidney to metabolic acidosis induced by ammonium chloride.

O rim desempenha um papel central na manutenção da composição de fluidos corporais através do controlo de água, NaCl, ácido-base, e reabsorção do soluto e excreção, respectivamente. O estudo foi realizado para investigar o papel fisiológico de tiamina na regulação da resposta renal à acidose metabólica induzida por NH4Cl em ratos machos adultos. Para esta experiência, foram utilizados cinquenta ratos. Eles foram divididos em cinco grupos. Os ratos de controlo receberam dieta basal; os ratos alimentados com dieta basal misturado com NH 4 Cl (4 g NH 4 Cl / 100 g de dieta) para induzir uma grande taxa de acidose metabólica , os ratos alimentados com dieta suplementada basal com tiamina (600 mg / kg de ração), e ratos alimentados com dieta suplementada basal com tiamina antes e após a indução de acidose metabólica por NH4Cl durante 14 dias. Os resultados mostraram que os níveis plasmáticos de cloreto, ureia e creatinina foram significativamente elevados em acidose metabólica induzida por NH4Cl. A suplementação de tiamina em doses elevadas antes ou após a indução aumentou os valores de cloreto. O fornecimento de dietas suplementadas com valores de tiamina modulado o sódio plasmático e bicarbonato. A suplementação com vitamina B1 como agente preventivo restaurou significativamente estas alterações para aproximar o valor de controle e, quando utilizado como agente curativo melhorou os níveis de creatinina e uréia plasmática. Os níveis de pH e de potássio na urina foram reduzidos significativamente em ratos acidóticos metabólicos, quando comparado com todos os grupos experimentais. os níveis de amônia e de aldosterona urinário foram reduzidos pela suplementação de tiamina como agente de proteção. A suplementação com vitamina B1 como agentes preventivos e curativos, restaurou os parâmetros afetados e regulou a resposta do rim a acidose metabólica induzida por cloreto de amónio.

Avaliação clínica, laboratorial e anatomopatológica do sistema urinário de ovinos confinados com ou sem suplementação de cloreto de amônio / Clinical, laboratory and anatomopathological evaluation of the urinary system of feedlot sheep with or without ammonium chloride supplementation

A acidificação urinária com cloreto de amônio (CA) é um método preventivo eficiente em urolitíase obstrutiva em ovinos. Os objetivos deste estudo com ovinos confinados, que receberam dieta concentrada com elevado teor proteico, foram: verificar o efeito da dieta sobre a formação de urólitos e o desenvolvimento da doença; analisar as características macroscópicas e histopatológicas do sistema urinário; relacionar os achados clínicos, laboratoriais e necroscópicos com a presença de urólitos. Utilizaram-se 60 ovinos machos: grupo CA (n=40), 400 mg/kg CA/dia, tratados via oral, por 42 dias consecutivos; grupo-controle (n=20), não tratado. Determinaram-se sete momentos de colheita de amostras com intervalos de sete dias, no total de 56 dias de confinamento. Encontraram-se microcálculos na pelve renal em cinco animais de ambos os grupos. As lesões renais microscópicas mais relevantes foram congestão vascular e necrose tubular. Concluiu-se que a dieta rica em concentrado provocou lesão renal em ambos os grupos, embora sem alterar a função renal, o que foi comprovado em testes pela ureia e creatinina séricas. O cloreto de amônio fornecido ao grupo CA não impediu a calculogênese, mas reduziu sua prevalência em relação ao grupo-controle. Os ovinos do grupo-controle tiveram maior comprometimento renal, pela alta incidência de cristalúria e pela necrose tubular, induzidas pelo consumo da dieta rica em grãos.

The urinary acidification with ammonium chloride (AC) is an efficient preventive method for urolithiasis in sheep. The objectives of this study with feedlot sheep receiving concentrated diet with high protein content were (1) to verify the effect of diet on urolith formation and development of the disease, (2) to analyze the macroscopic and histopathological characteristics of the urinary system, and (3) to relate the clinical, laboratory and necropsy findings with the presence of uroliths. Sixty male sheep were used: AC group (n=40), 400mg/kg AC/day, orally treated for 42 consecutive days, and control group (n=20), untreated. Seven times were determined for sampling with a seven-day interval, totaling 56 days of feedlot. Small uroliths were found in the renal pelvis of five sheep in both groups. The most relevant microscopic renal lesions were vascular congestion and tubular necrosis. It was concluded that the highly concentrated diet caused renal injury in both groups, without changing the renal function, what was proven by laboratory tests of urea and creatinine. Ammonium chloride provided to the CA group did not prevent urolith formation, but reduced its prevalence in comparison with the control group. Sheep of the control group had increased kidney damage, which resulted in higher incidence of crystalluria and tubular necrosis induced by the consumption of a diet rich in grains.

Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System

BACKGROUND: We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. METHODS: Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMerieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. RESULTS: The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. CONCLUSIONS: This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.

Inclusão de cloreto de amônia na dieta pré-parto de ovelhas leiteiras e sua influência nos metabolismos do cálcio e energético / Inclusion of ammonium chloride in the pre-partum diet in dairy sheep and its influence on calcium and energetic metabolism

Esta pesquisa foi realizada com o objetivo de avaliar o efeito da inclusão de cloreto de amônia na dieta pré-parto de ovelhas leiteiras sobre o pH urinário, cálcio sérico e metabolismo energético dos animais. Foram utilizadas vinte fêmeas ovinas da raça Lacaune, separadas em dois grupos: o Grupo Controle (GC, n=10) que recebeu a dieta pré-parto sem adição de cloreto de amônia e o Grupo Tratado (GT, n=10) que recebeu a dieta pré-parto com a inclusão de 0,4% de cloreto de amônia do total de matéria seca (MS) consumida por animal/dia. As coletas de urina e sangue ocorreram aos 120 (M0), 127 (M1) e 135 (M2) dias de gestação. Foram avaliados o pH urinário e os níveis séricos de cálcio, cetonas, glicose e colesterol. Houve redução no pH urinário no GT nos dias 127 e 135 de gestação. Os valores de cetonas séricas foram inferiores no GT no dia 135 de gestação enquanto que os níveis de cálcio e colesterol foram superiores neste grupo no mesmo momento tempo avaliado. Não houve diferença nos valores de glicose entre os grupos. Desta forma, conclui-se que a inclusão de 0,4% de cloreto de amônia do total de matéria seca consumida por animal/dia na dieta pré-parto de ovelhas leiteiras foi eficiente na manutenção da calcemia neste período e que esta pode estar associada à não redução do status energético em ovelhas gestantes.

This research was done to evaluate the effect of inclusion of ammonium chloride in the pre-partum diet in dairy sheep over the urinary pH and in calcium and energetic metabolism of these animals. Twenty ewe of Lacaune breed were separated into two groups, the Control Group (CG, n = 10) that received pre-partum diet without ammonium chloride and Treated Group (TG, n = 10) that received the diet pre-partum with 0.4% of ammonium chloride in total dry matter (DM) consumed per animal/day. The samples of urine and blood were collected in 120 (M0), 127 (M1) and 135 (M2) days of gestation. The parameters assessed were the urine pH and serum levels of calcium, ketones, glucose and cholesterol. In the urinary pH assay occurred a reduction in the GT on 127 and 135 days of gestation. The values were lower in the serum ketones in GT on 135 of gestation on the day whereas calcium levels and cholesterol levels were higher in this group at the same experimental time. There was no difference in glucose values between groups. Thus, it is concluded that inclusion of 0.4% of ammonium chloride to the total dry matter intake per animal/day in the pre-partum diet of milk sheep was effective in maintaining calcium levels over this period. And yet, this maintenance of calcium levels may be associated with no reduction in energy status in pregnant ewes.

The Effects of Temperature and Nutritional Conditions on Mycelium Growth of Two Oyster Mushrooms (Pleurotus ostreatus and Pleurotus cystidiosus)

The influences of temperature and nutritional conditions on the mycelium growth of oyster mushroom Pleurotus ostreatus (PO) and Pleurotus cystidiosus (PC) were investigated in laboratory experiment during the summer season of 2014. The results of the experiment indicated that potato dextrose agar (PDA) and yam dextrose agar (YDA) were the most suitable media for the mycelium growth of oyster mushroom PO while four media (PDA, YDA, sweet potato dextrose agar, and malt extract agar medium) were not significantly different in supporting mycelium growth of oyster mushroom PC. The optimal temperature for mycelium growth of both oyster mushroom species was obtained at 28degrees C. Mycelium growth of oyster mushroom PO was improved by carbon sources such as glucose, molasses, and at 1~5% sucrose concentration, mycelium colony diameter of mushroom PO was achieved the highest value. Whereas glucose, dextrose, and sucrose as carbon sources gave the good mycelium growth of oyster mushroom PC, and at 1~3% sucrose concentration, mycelium colony diameter of PC was achieved the maximum value. Ammonium chloride concentrations at 0.03~0.09% and 0.03~0.05% also gave the greatest values in mycelium colony diameter of mushroom PO and PC. Brown rice was found to be the most favourable for mycelium growth of two oyster mushroom species. In addition, sugarcane residue, acasia sawdust and corn cob were selected as favourable lignocellulosic substrate sources for mycelium growth of both oyster mushrooms.

Alterações ultrassonográficas renais e vesicais de ovinos confinados e suplementados com cloreto de amônio / Renal and bladder ultrasound changes in feedlot sheep and supplemented with ammonium chloride

A incidência da urolitíase obstrutiva em ovinos é elevada, principalmente em machos confinados, tanto para produção de carne, quanto reprodutores de alto valor genético. A acidificação urinária é um dos métodos para prevenção desta enfermidade e pode ser realizada de forma eficaz com a suplementação de cloreto de amônio (CA) na dieta. Utilizaram-se 100 ovinos, machos não castrados, mestiços (Ile de France X White Dorper), confinados, com idade aproximada de três meses. Constituíram-se três grupos experimentais: Grupo 21CA (n=40) que recebeu 400mg/kg/PV de cloreto de amônio/animal/dia, por 21 dias consecutivos; Grupo 42CA (n=40) que foi suplementado com 400mg/kg/PV de cloreto de amônio/animal/dia, por 42 dias consecutivos; Grupo controle (n=20), que não recebeu CA. A alimentação consistiu de ração total, composta por 15% de feno triturado e 85% de concentrado, água e sal mineral ad libitum. Após 14 dias de adaptação à alimentação e ao ambiente, os Momentos (M) de avaliação clínica, colheita de sangue e exame ultrassonográfico foram realizados com intervalo de sete dias, sendo M0 (imediatamente antes do início do tratamento com cloreto de amônio), M1 (sete dias após), M2 (14 dias após), M3 (21 dias após o início do tratamento e suspensão do cloreto de amônio em Grupo 21CA), M4 (28 dias após), M5 (35 dias após) e M6 (42 dias após), totalizando 56 dias de confinamento. As dosagens de ureia e creatinina não evidenciaram alteração na função renal, embora a ureia estivesse acima dos valores de referência para espécie ovina. Observaram-se imagens ultrassonográficas compatíveis com cálculos vesicais e dilatação de pelve renal. No Grupo 21CA, 15% (6/40) dos animais apresentaram cálculos vesicais; no Grupo 42CA, 5% (2/40); e no Grupo controle, 20% (4/20) dos cordeiros. Visibilizaram-se também imagens sugestivas de sedimentos e cristais em 31% (31/100) dos animais examinados. A ultrassonografia permitiu a visibilização de alterações renais e vesicais, porém não relacionados ao quadro clínico de urolitíase obstrutiva, revelando-se como um exame complementar de grande relevância para o diagnóstico precoce de alterações no sistema urinário de ovinos.(AU)

The incidence of obstructive urolithiasis in sheep is high, especially in feedlot males, both for meat production, or the breeder of high genetic value. The urinary acidification is one of the methods for preventing this disease and can be performed efficiently with supplementation of ammonium chloride (AC) in the diet. It was used 100 male lambs, in a feedlot, crossbred (Ile de France X White Dorper), aged approximately three months. It was constituted three groups: Group 21AC (n=40) that received 400mg/kg/PV of ammonium chloride/animal/day for 21 consecutive days, the time of discontinuation of the urinary acidifiers (M3) and continued clinical follow until the end of the experiment (M6); Group 42AC (n=40), that received 400mg/kg/PV of ammonium chloride/animal/day for 42 consecutive days, Group control (n=20), that did not receive ammonium chloride throughout the experimental period. The feed consisted of total dry matter, composed of 15% ground hay and 85% concentrate, water and mineral salts ad libitum. After 14 days of adaptation to food and the environment, the moments (M) for clinical evaluation, and blood collection ultrasound examinations were performed with an interval of seven days, and M0 (immediately before the beginning of the treatment with ammonium chloride), M1 (seven days), M2 (14 days after), M3 (21 days after initiation of treatment and suspension of ammonium chloride in Group 21CA), M4 (28 days), M5 (35 days), and M6 (42 days), amounting to 56 days of feedlot. The serum urea and creatinine showed no change in renal function, although the urea was above the reference values for sheep. There were compatible ultrasound images with bladder stones and dilatation of the renal pelvis. In Group 21AC, 15% (6/40) of the animals had bladder stones; in Group 42AC 5% (2/40); and in Group control, 20% (4/20) of the lambs. It was visualized suggestive images of sediment and crystals in 31% (31/100) of examined animals. Ultrasonography allowed visualization of kidney and bladder abnormalities, which were not related to clinical symptoms of obstructive urolithiasis, appearing as an examination complement of great importance for the early detection of changes in the urinary system of sheep.(AU)

Efeito da suplementação de cloreto de amônio sobre os equilíbrios eletrolítico e ácido-básico e o pH urinário de ovinos confinados / Effect of supplementation of ammonium chloride on electrolyte and acid-base balances and urinary pH of feedlot sheep

A incidência da urolitíase obstrutiva em ovinos é elevada, principalmente em machos confinados, tanto para produção de carne, quanto reprodutores de alto valor genético. A acidificação urinária é um dos métodos para prevenção desta enfermidade e pode ser realizada de forma eficaz com a suplementação de cloreto de amônio na dieta, que pode propiciar a instalação de acidose metabólica. A hemogasometria avalia o equilíbrio ácido-básico sanguíneo de forma prática e fácil. Neste trabalho, avaliou-se o efeito do cloreto de amônio sobre o equilíbrio ácido-básico e eletrolítico de ovinos em confinamento para quantificar a acidose metabólica desenvolvida. Utilizaram-se 100 ovinos, machos, confinados, com idade aproximada de três meses. Constituíram-se três grupos experimentais: Grupo I (n=40), recebeu 400mg/kg/PV de cloreto de amônio/animal/dia por 21 dias consecutivos, momento da interrupção da administração do acidificante urinário (M3) e continuidade do acompanhamento clinico até o final do experimento (M6); Grupo II (n=40), 400mg/kg/PV de cloreto de amônio/animal/dia por 42 dias consecutivos; Grupo III (n=20), não recebeu cloreto de amônio durante todo o período do experimento. Os Momentos (M) de colheita de amostras e avaliação clínica foram estabelecidos com intervalo de sete dias, sendo M0 (imediatamente antes do início do tratamento com cloreto de amônio), M1 (sete dias após), M2, M3, M4, M5 e M6, totalizando 56 dias de confinamento. A alimentação consistiu de ração total, composta por 15% de feno triturado e 85% de concentrado, água e sal mineral ad libitum. Após adaptação de 15 dias à dieta de confinamento, colheram-se de todos os animais amostras de urina para mensuração do pH, e sangue venoso para hemogasometria, nos diferentes momentos...

The incidence of obstructive urolithiasis in sheep is high, especially in feedlot males, both for meat production, or the breeder of high genetic value. The urinary acidification is one way to prevent this disease and can be performed effectively supplementation with ammonium chloride in the diet, which may facilitate the installation of metabolic acidosis. The blood gas analysis evaluates the acid-base balance of blood in a practical and easy way. In this study, it was evaluated the effect of ammonium chloride on acid-base and electrolyte in feedlot sheep blood gas analysis to determine the occurrence of metabolic acidosis. It was used 100 male lambs, in a feedlot, aged approximately three months. It was constituted three groups: Group I (n=40) that received 400mg/kg/PV of ammonium chloride/animal/day for 21 consecutive days, the time of discontinuation of the urinary acidifiers (M3) and continued clinical follow until the end of the experiment (M6); Group II (n=40), that received 400mg/kg/PV of ammonium chloride/animal/day for 42 consecutive days, Group III (n=20), that did not receive ammonium chloride throughout the experimental period. The moments (M) of samples and clinical assessment were established on seven days of interval, M0 (immediately before the beginning of the treatment with ammonium chloride), M1 (seven days after), M2, M3, M4, M5 and M6, totalizing 56 days of feedlot. The feed consisted of a total mixed ration consisting of 15% of ground hay and 85 % of concentrate, water and mineral salt ad libitum. After 15 days of adaptation to the diet of feedlot, urine samples for measurement of pH, and venous blood for blood gas analysis were collected from all animals at different moments. The urinary acidification was maintained as was the administration of ammonium chloride in GI and GII. The values of Na+ and K+ remained within the normal range for the species...

Diosmin Reduces Calcium Oxalate Deposition and Tissue Degeneration in Nephrolithiasis in Rats: A Stereological Study

PURPOSE: Kidney stones (nephrolithiasis) are a widespread disease. Thus, blocking stone formation and finding new therapeutic methods is an important area of study. Diosmin (a major component of the bile) is known to have antioxidant as well as renoprotective effects. The present investigation aimed to evaluate the effect of diosmin on renal tissue protection in rats with ethylene glycol-induced nephrolithiasis. MATERIALS AND METHODS: The rats were randomly divided into three groups. Group one (control) did not receive any treatments. In groups two and three, nephrolithiasis was induced by 2.5% (V/V) ethylene glycol + 2.5% (W/V) ammonium chloride (2 mL/d). The second and the third groups received distilled water or diosmin (80 mg/kg/d) by gavage for 21 days. RESULTS: Stereological estimation of the renal structures revealed that the average volume of calcium oxalate (CaOx) in the nephrolithiasis+diosmin rats was -63% less than in the rats with untreated nephrolithiasis (p<0.01). The volume of the glomeruli, proximal and distal convoluted tubules, Henle's loop, collecting ducts, and vessels was reduced -32% to 58% after the induction of nephrolithiasis (p<0.001). In the nephrolithiasis+diosmin rats, on average, -70% to 96% of the glomeruli, proximal convoluted tubules, Henle's loop collecting ducts, and vessels remained intact (p<0.01). Degeneration of the cortical tissue was 5-fold that of the medulla. In the nephrolithiasis+diosmin rats, degeneration in the renal cortical tissue and medulla was reduced -70% and 44%, respectively, compared with that in the untreated nephrolithiasis group (p<0.01). CONCLUSIONS: Diosmin reduces CaOx deposition and the degeneration of glomeruli and tubules in a rat model of nephrolithiasis.

A Randomized, Open Label, Multicentre Study to Evaluate the Safety and Efficacy of Cough Mixture CS1 (Pholcodeine and Promethazine - Tixylix) Versus CS2 (Noscapine, Ammonium Chloride, Sodium Citrate) in Treatment of Children with Dry Cough.

The objective of this study was to compare the efficacy and safety of cough mixture containing pholcodeine and promethazine - Tixylix (CS1) to a cough mixture which has noscapine, ammonium chloride, and sodium citrate (CS2) as its constituents in treatment of children suffering from dry cough. A total of 208 patients were enrolled at 4 sites. Of these, 179 (94 receiving CS1 and 99 receiving CS2) completed the study. Results of this study suggest that both the cough mixtures were comparable as per evaluation of their primary parameters. According to global assessment for efficacy and tolerability by parents on Day 7, Group CS1 performed better than CS2. It was also observed that no AE was reported in Group CS1 as compared to 2 AEs in Group CS2. To conclude, cough mixture combination of pholcodeine and promethazine - Tixylix exhibited efficacy and safety that was comparable with cough mixture which has noscapine, ammonium chloride, and sodium citrate. It was proven to be efficacious, safe and well tolerated in the select population.

Altered Regulation of Renal Acid Base Transporters in Response to Ammonium Chloride Loading in Rats

The role of the kidney in combating metabolic acidosis has been a subject of considerable interest for many years. The present study was aimed to determine whether there is an altered regulation of renal acid base transporters in acute and chronic acid loading. Male Sprague-Dawley rats were used. Metabolic acidosis was induced by administration of NH4Cl for 2 days (acute) and for 7days (chronic). The serum and urinary pH and bicarbonate were measured. The protein expression of renal acid base transporters [type 3 Na+/H+ exchanger (NHE3), type 1 Na+/HCO3- cotransporter (NBC1), Na-K+ ATPase, H(+)-ATPase, anion exchanger-1 (AE-1)] was measured by semiquantitative immunoblotting. Serum bicarbonate and pH were decreased in acute acid loading rats compared with controls. Accordingly, urinary pH decreased. The protein expression of NHE3, H(+)-ATPase, AE-1 and NBC1 was not changed. In chronic acid loading rats, serum bicarbonate and pH were not changed, while urinary pH was decreased compared with controls. The protein expression of NHE3, H(+)-ATPase was increased in the renal cortex of chronic acid loading rats. These results suggest that unaltered expression of acid transporters combined with acute acid loading may contribute to the development of acidosis. The subsequent increased expression of NHE3, H(+)-ATPase in the kidney may play a role in promoting acid excretion in the later stage of acid loading, which counteract the development of metabolic acidosis.

The Patch Test as a Useful Tool for Avoiding Suspected Allergens in Patients with Hand Eczema / 대한피부과학회지

BACKGROUND: Hand eczema is a common skin disease in the general population. The etiology of hand eczema is obscure and many causative factors have been proposed. However, there are only a few reported studies of the relevance of contact allergy in hand eczema. Objective: The purpose of this study was to evaluate the diagnostic value of the patch test for patients with hand eczema. METHODS: We analyzed the clinical characteristics and the results of the patch tests of the 37 patients with hand eczema and we then compared these with the clinical subtypes. RESULTS: 26 patients (70.3%) showed a positive test to one or more allergens. The common allergens were nickel sulfate (35.1%), mercury ammonium chloride (21.6%), and cobalt chloride (18.9%). The positive rates for a patch test were 82.4% for the vesicular form, 77.8% for the fissured form, 20.0% for the hyperkeratotic form and 66.7% for pompholyx, respectively. We found more significant improvement of the clinical symptoms in the vesicular group (57.1%) than that in the non-vesicular group (9.3%) after avoiding the verified allergens. CONCLUSION: This study shows that the patch test is a useful tool for the detection of contact allergens and it must be performed for the patients with hand eczema, and especially for those patients with the vesicular type.

The Patch Test as a Useful Tool for Avoiding Suspected Allergens in Patients with Hand Eczema / 대한피부과학회지

BACKGROUND: Hand eczema is a common skin disease in the general population. The etiology of hand eczema is obscure and many causative factors have been proposed. However, there are only a few reported studies of the relevance of contact allergy in hand eczema. Objective: The purpose of this study was to evaluate the diagnostic value of the patch test for patients with hand eczema. METHODS: We analyzed the clinical characteristics and the results of the patch tests of the 37 patients with hand eczema and we then compared these with the clinical subtypes. RESULTS: 26 patients (70.3%) showed a positive test to one or more allergens. The common allergens were nickel sulfate (35.1%), mercury ammonium chloride (21.6%), and cobalt chloride (18.9%). The positive rates for a patch test were 82.4% for the vesicular form, 77.8% for the fissured form, 20.0% for the hyperkeratotic form and 66.7% for pompholyx, respectively. We found more significant improvement of the clinical symptoms in the vesicular group (57.1%) than that in the non-vesicular group (9.3%) after avoiding the verified allergens. CONCLUSION: This study shows that the patch test is a useful tool for the detection of contact allergens and it must be performed for the patients with hand eczema, and especially for those patients with the vesicular type.

Ammonia induces upregulation of aquaporin-4 in neocortical astrocytes of rats through the p38 mitogen-activated protein kinase pathway / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Astrocyte swelling is an important consequence of hepatic encephalopathy, and aquaporin-4 has been reported to play a vital role in this swelling. Ammonia causes astrocyte swelling and is also known to modulate aquaporin-4 expression in the astrocyte foot processes. The purpose of this study was to explore the mechanism of ammonia-induced aquaporin-4 expression, which has been suggested to involve the p38 mitogen-activated protein kinase pathway.</p><p><b>METHODS</b>We exposed cultured astrocytes to ammonium chloride, an in vitro model of hepatic encephalopathy. The purity of cultured astrocytes was evaluated by fluorescent glial fibrillary acidic protein labeling; cell morphology was assessed by light microscopy; the expression of aquaporin-4, phospho-p38, and p38 were detected by Western blotting analysis. Statistical analysis was performed by one-way factorial analysis of variance, and the relationship between variables was calculated by linear regression using SPSS version 13.0 program for Windows (SPSS, Chicago, IL, USA).</p><p><b>RESULTS</b>The purity of cultured astrocytes was (96.6 +/- 1.4)%. Astrocytes swelled significantly when exposed to 5 mmol/L ammonium chloride for 24 hours as compared to non-exposed astrocytes. Co-treatment with 10 micromol/L SB203580 (an inhibitor of p38) attenuated the degree of ammonium chloride induced astrocyte swelling. Western blotting analysis revealed that the expression levels of phospho-p38 and aquaporin-4 in ammonium chloride treated cells were significantly increased relative to the control group (P < 0.001); SB203580 co-treatment inhibited the increased expression of phospho-p38 and aquaporin-4 relative to the ammonium chloride treated group (P = 0.002 and P = 0.015 respectively). The phosphorylation of p38 and upregulation of aquaporin-4 were highly correlated (r = 0.909). There were no significant differences in total p38 expression among the groups (P = 0.341).</p><p><b>CONCLUSIONS</b>Ammonium chloride induced upregulation of aquaporin-4 in astrocytes is regulated by the p38 mitogen-activated protein kinase pathway. Inhibiting p38 activation prevented ammonium chloride induced aquaporin-4 protein upregulation.</p>

Medium optimization for enhanced production of carbonyl reductase by Candida tropicalis 104 by response surface methodology / 生物工程学报

Using response surface method, we optimized the medium for the asymmetric whole cell biotransformation by Candida tropicalis 104. This strain was used for microbial reduction of 1-[3,5-bis(trifluoromethyl)phenyl] ethanone to (S)-l-[3,5-bis(trifluoromethyl)phenyl] alcohol, with enantiomeric excess(e.e.) reached more than 99.9%. Fractional factorial design was used to evaluate the effects of medium components on carbonyl reductase activity of Candida tropicalis 104. Yeast extract, glucose and NH4Cl were the most important factors among six tested variables that influence the enzyme activity for the biotransformation process. Based on the experimental results, the path of steepest ascent was undertaken to approach the optimal region of these factors. Central composite design and response surface analysis were subsequently employed for further optimization. The optimal medium for Candida tropicalis 104 was composed of (in g/L): glucose 47.14, yeast extract 13.25, NH4Cl 2.71, MgSO4.7H2O 0.4, KH2PO4 1, K2HPO4 1. Under the optimum conditions, the maximum enzyme activity of 852.75 U/L in theory and 851.13 U/L in the experiment were obtained, with an increase of 65.2% compared to the original medium components.

Standard Patch Test Results in Patients with Contact Dermatitis / 대한피부과학회지

BACKGROUND: A number of skin manifestations in patients with allergic contact dermatitis result from sensitization with specific allergens. The patch test is a primary diagnostic tool for identifying allergens causing allergic contact dermatitis. OBJECTIVE: We identified the results of patch test with Korean standard series in patients with clinically diagnosed with contact dermatitis, and evaluated the specific relationship with each allergen. METHODS: During a 5-year period, 743 patients were examined with patch test of standard Korean series. The results of patch test were then evaluated. Location of the skin lesion, age, sex, and occupation of the patients were completely reviewed. RESULTS: Overall, 68.5% of patients had at least one positive reaction and 43.6% of patients had two or more positive reactions. Allergens most frequently causing positive reactions were nickel sulfate (27.1%), cobalt chloride (22.1%), potassium dichromate (17.9%), mercuric ammonium chloride (13.8%), and 4-phenyledediamine base (10.2%). Those patients in their 10s and 40s showed the highest rate of positive reaction. Metal allergens displayed higher positive rates than any other standard allergens. Of 201 patients who showed positive reaction to nickel, 37.8% and 27.3% of them also showed positive reaction to cobalt and chromium, respectively, representing concomitant reactions between metal-related antigens. In addition, polysensitization to cosmetics-related antigens including balsam peru and fragnance mix was evident. CONCLUSION: There was no significant difference in overall positive rate in patch test results compared with recent studies. However, we confirmed that metal-related allergens remain most common, and that multiple contact allergy were relatively common.