Prevalence of gastrointestinal parasitic infection in cows and buffaloes in Lower Dir, Khyber Pakhtunkhwa, Pakistan

Abstract Gastrointestinal (GI) Parasitic infection is a hot issue for cattle management. There is variation of GI parasites effects in sex, age of cattle, drinking water condition, nutrition, and severity of infection. Studies on prevalence of GI parasites among cattle population in Dir Lower are lacking. A total of 40 farms were selected randomly in six tehsil namely Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda ,Tehsil Lalqala, Tehsil Khall. Freshly cattle fecal samples were collected randomly from the selected farms during March 2018 till December 2018. Out of 314 buffaloes and cattle examined 58.59% (184/314) were positive for eggs, cyst/oocyst of one or more species of GI Parasites. The prevalence of parasitic infection was higher in Buffaloes 63.55% (75/118) as compared to Cow 55.61% (109/196) but the difference was not significant (p>0.05) Entamoeba,spp, Moniezia spp, Haemonchus spp and Coccidian spp were found in this study. The non-treated animals indicated the highest percentage of infection in cow 57.71% (101/175) and buffalo 68.13% (62/91).GI parasite prevalence in female animal were higher female cow 62.58% (87/139) and female buffalo 77.33% (58/75) as compared to male. But the difference is non-significant (p> 0.05) Yearling calves had the lower rate of GI parasitic infection than adults. Future investigations are necessary to evaluate the economic loss due to GI parasites in cattles.

Resumo A infecção gastrointestinal (GI) parasitária é um assunto importante para o manejo do gado. Existem variações nos efeitos dos parasitas GI quanto a sexo, idade do gado, condição da água potável, nutrição e gravidade da infecção. Faltam estudos sobre a prevalência de parasitas GI entre a população de gado em Lower Dir. Um total de 40 fazendas foi selecionado aleatoriamente em seis tehsil, nomeadamente Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda, Tehsil Lalqala, Tehsil Khall. Amostras fecais de gado fresco foram coletadas aleatoriamente das fazendas selecionadas de março de 2018 até dezembro de 2018. Dos 314 búfalos e bovinos examinados, 58,59% (184/314) foram positivos para ovos, cisto/oocisto de uma ou mais espécies de parasitas GI. A prevalência de infecção parasitária foi maior em Buffaloes 63,55% (75/118) em comparação com vaca 55,61% (109/196), mas a diferença não foi significativa (p > 0,05). Entamoeba spp, Moniezia spp, Haemonchus spp e Coccidian spp foram encontrados neste estudo. Os animais não tratados indicaram a maior porcentagem de infecção em vacas 57,71% (101/175) e búfalos 68,13% (62/91). A prevalência do parasita GI em fêmeas foi maior em vacas fêmeas 62,58% (87/139) e búfalas 77,33% (58/75) em comparação ao masculino. Mas a diferença não é significativa (p > 0,05). A modelagem linear geral mostrou que o tratamento do animal estava significativamente relacionado com a prevalência de parasitas GI. Bezerros de um ano tiveram a maior taxa de infecção parasitária gastrointestinal. Futuras investigações são necessárias para avaliar a perda econômica devido aos parasitas GI em bovinos.

Prevalence of gastrointestinal parasitic infection in cows and buffaloes in Lower Dir, Khyber Pakhtunkhwa, Pakistan.

Gastrointestinal (GI) Parasitic infection is a hot issue for cattle management. There is variation of GI parasites effects in sex, age of cattle, drinking water condition, nutrition, and severity of infection. Studies on prevalence of GI parasites among cattle population in Dir Lower are lacking. A total of 40 farms were selected randomly in six tehsil namely Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda ,Tehsil Lalqala, Tehsil Khall. Freshly cattle fecal samples were collected randomly from the selected farms during March 2018 till December 2018. Out of 314 buffaloes and cattle examined 58.59% (184/314) were positive for eggs, cyst/oocyst of one or more species of GI Parasites. The prevalence of parasitic infection was higher in Buffaloes 63.55% (75/118) as compared to Cow 55.61% (109/196) but the difference was not significant (p>0.05) Entamoeba,spp, Moniezia spp, Haemonchus spp and Coccidian spp were found in this study. The non-treated animals indicated the highest percentage of infection in cow 57.71% (101/175) and buffalo 68.13% (62/91).GI parasite prevalence in female animal were higher female cow 62.58% (87/139) and female buffalo 77.33% (58/75) as compared to male. But the difference is non-significant (p> 0.05) Yearling calves had the lower rate of GI parasitic infection than adults. Future investigations are necessary to evaluate the economic loss due to GI parasites in cattle's.

Assessment of ecto and endo parasites of Schizothorax plagiostomus inhabiting river Panjkora, Khyber Pakhtunkhwa, Pakistan / Avaliação de ecto e endoparasitas de Schizothorax plagiostomus habitantes do rio Panjkora, Khyber Pakhtunkhwa, Paquistão

Abstract Fish provides main source of high quality protein to more than one billion people in the world. Fish parasites directly affect the productivity of fish and indirectly on human health. This research was aimed to assess the helminth parasites in Schizothorax plagiostomus (the snow trout) from river Panjkora, KP, Pakistan. A total of 88 fish samples (n= 88) were collected from the upper, middle and lower regions of the river through cast nets, hand nets and hooks. All the collected fish samples were examined in the Parasitology Laboratory, Department of Zoology, University of Malakand for helminth parasites during July 2018 to February 2019.Among these fishes nematodes, trematodes, cestodes and acanthocephalan were isolated and identified. Overall prevalence of the fish parasites was 60.22% (53/88). The intensity of the parasite was observed from 1.7% to 4.61%. Highest prevalence was reported in summer season 86.36% while lowest in the winter season 36.36%. This study shows that summer season affects the fish business and result in poor quality fish meat with risk of zoonotic diseases. Adults were highly infected 82.25% while no infection was found in juvenile specimens. Female fish samples had higher prevalence 68.22% than males 48.22%. Fishes of the lower reaches had highest prevalence 60.22% than the upper reaches 40.0%. Fish samples with maximum length and weight were highly 76.92% infected than small sized fish with low body weight. Present study addresses that Rhabdochona schizothoracis in the intestine and Diplozoon paradoxum in gills of snow trout fish has a long term relationship and call as a natural infection in cyprinids and it is zoonotic threat to human.

Resumo O peixe é a principal fonte de proteína de alta qualidade para mais de um bilhão de pessoas no mundo. Contudo, parasitas afetam diretamente a produtividade de peixes e, indiretamente, a saúde humana. Esta pesquisa teve como objetivo avaliar os parasitas helmintos em Schizothorax plagiostomus (truta da neve) do rio Panjkora, Khyber Pakhtunkhwa, Paquistão. Foram coletadas 88 amostras de peixes (n = 88) dos trechos alto, médio e baixo do rio por meio de redes de arrasto, redes de mão e varas de pesca. Todas as amostras de peixes coletadas foram examinadas no Laboratório de Parasitologia do Departamento de Zoologia, da Universidade de Malakand, quanto a parasitas helmintos durante o período de julho de 2018 a fevereiro de 2019. Entre esses peixes, nematódeos, trematódeos, cestóideos e acantocéfalos foram isolados e identificados. A prevalência geral de parasitas de peixes foi de 60,22% (53/88). A intensidade do parasita foi observada de 1,7 a 4,61%. A maior prevalência (86,36%) foi relatada no verão, enquanto a menor (36,36%), no inverno. Este estudo mostra que a temporada de verão afeta o ramo da pesca e resulta em carne de baixa qualidade, com risco de doenças zoonóticas. Os adultos foram altamente infectados (82,25%), enquanto nenhuma infecção foi encontrada em jovens. As amostras de peixes do sexo feminino apresentaram maior prevalência (68,22%) do que as do sexo masculino (48,22%). Os peixes coletados no trecho baixo do rio tiveram maior prevalência (60,22%) do que os do trecho alto (40%). Amostras de peixes com comprimento e peso máximos foram altamente infectadas (76,92%) do que peixes de pequeno porte e baixo peso corporal. O presente estudo aborda que Rhabdochona schizothoracis no intestino e Diplozoon paradoxum em brânquias de peixes de truta da neve possuem uma estreita relação que causa uma infecção natural em ciprinídeos, sendo uma ameaça zoonótica ao ser humano.

Evaluation of ecto and endo parasitic fauna of Schizothorax plagiostomus inhabitants of river Swat, Khyber PakhtunKhwa, Pakistan / Avaliação da fauna ecto e endoparasitária de Schizothorax plagiostomus habitantes do rio Swat, Khyber Pakhtunkhwa, Paquistão

Abstract Fish is consumed as a rich and cheaper source of white meat and proteins all over the world. Fish farming is the leading source of income generation throughout the world. The present research study was conducted with aims to investigate the ecto and endo parasitic fauna of S. plagiostomus (snow trout) in River Swat. A total of 96 fish samples were collected on monthly basis from July-2018 to February-2019, from upper, middle and lower reaches. Fish samples were collected with the help of cast net, hand net and fishing rods. Local fishermen and experts help were also sought out for fish collection. 61 out of 96 fishes were found infected by helminth parasites. The total prevalence and intensity of 63.54% and 2.90 was observed respectively during data analysis. Highest monthly prevalence of 83.33% was recorded during July and August, while lowest prevalence of 33.33% was noted during February. Five species of helminths parasites were detected from S. plagiostomus, of which two were trematodes (Rhabdochona shizothoracis and Rhabdochona species), one species of Monogenia (Diplozoon paraddoxum), one species of Cestodes (Nippotaenia species), and one species of Acanthocephalan (Neoechynorhynchus devdevi). Highest parasite wise prevalence, intensity and relative density of 21.87%, 4.09 and 0.895 was noted for R. schizothoracis while the lowest prevalence, intensity and relative density of 4.16%, 1.25 and 0.052 was noted for N. devdevi. Highest infection of 76.08% was observed in adults host while lowest, 40% infestation rate was observed in young fish samples. 73.68% prevalence was observed in female hosts while only 56.90% prevalence was observed in male individuals. Higher infection (83.33%) was noticed during summer season, while lowest infection (44.44%) was observed during winter season. Similarly 71.79% fishes were found infected that were collected from lower reaches of the River Swat than the fish samples collected from upper reaches (52%).

Resumo O peixe é consumido mundialmente por ser uma fonte rica e barata de carne branca e proteínas. A piscicultura é a principal fonte de geração de renda em todo o mundo. O presente estudo foi conduzido com o objetivo de investigar a fauna ecto e endoparasitária de S. plagiostomus (truta da neve) no rio Swat. Foram coletadas mensalmente 96 amostras de peixes, no período de julho de 2018 a fevereiro de 2019, nos trechos alto, médio e baixo com auxílio de redes de arrasto, redes de mão e varas de pesca. Pescadores locais e especialistas também ajudaram na coleta de peixes. Dos 96 peixes encontrados, 61 estavam infectados por parasitas helmintos. A prevalência de 63,54% e a intensidade total de 2,90 foram observadas durante a análise dos dados. A maior prevalência mensal de 83,33% foi registrada nos meses de julho e agosto, enquanto a menor, de 33,33%, em fevereiro. Cinco espécies de parasitas helmintos foram detectadas de S. plagiostomus, dos quais dois eram trematódeos (espécies Rhabdochona shizothoracis e Rhabdochona), uma espécie de monogenia (Diplozoon paradoxum), uma espécie de cestóideos (espécie Nippotaenia) e uma espécie de acantocéfalo (Neoechynorhynchus devdevi). As maiores prevalências de parasitas (21,87%), intensidade (4,09) e densidade relativa (0,895) foram observadas em R. schizothoracis, enquanto as menores prevalências de parasitas (4,16%), intensidade (1,25) e densidade relativa (0,052), em N. devdevi. A infecção mais alta (76,08%) foi constatada em adultos hospedeiros, ao passo que a taxa de infestação mais baixa (40%) foi encontrada em amostras de peixes jovens. A prevalência de 73,68% foi verificada em hospedeiros do sexo feminino, enquanto a de 56,90%, em indivíduos do sexo masculino. Maior infecção (83,33%) foi observada durante a temporada de verão, e a menor infecção (44,44%), durante o inverno. Da mesma forma, 71,79% dos peixes encontrados infectados foram coletados no trecho baixo do rio Swat, enquanto 52%, no trecho alto.

Glycoamino Acid Analogues of the Thomsen-Friedenreich Tumor-Associated Carbohydrate Antigen: Synthesis and Evaluation of Novel Antiproliferative Factor Glycopeptides.

Glycoamino acid analogues of the Thomsen-Friedenreich antigen disaccharide, where the 4' and 4″ hydroxyl groups were substituted with fluorine or hydrogen, were synthesized and incorporated into the asialylated antiproliferative factor (as-APF), a biologically active form of APF, a glycopeptide found in the urine of patients with interstitial cystitis. Various strategies were employed to incorporate the fluorine atom at the 4-positions of either the galactose or N-acetylgalactosamine unit of the disaccharide antigen, based on stereochemistry and reactivity. These glycopeptides were evaluated in antiproliferative assays on both primary normal bladder epithelial cells and T24 bladder carcinoma cells. Unlike many previously published substitutions to APF, mono-4'-fluorination of the GalNAc residue did not affect the activity, whereas fluoro-derivatives of the galactose 4″-position or both 4' and 4″ hydroxyls showed a reduced potency relative to the monosubstituted GalNAc derivative. A fourth compound where the 4″ position of galactose was deoxygenated showed a lower potency than the parent and monosubstituted compounds. These results suggest that specific substitutions in the sugar moieties in the APF can be tolerated, and the glycomimetic design of APF analogues can include fluorine in the GalNAc sugar of the disaccharide.

Assessment of ecto and endo parasites of Schizothorax plagiostomus inhabiting river Panjkora, Khyber Pakhtunkhwa, Pakistan

Abstract Fish provides main source of high quality protein to more than one billion people in the world. Fish parasites directly affect the productivity of fish and indirectly on human health. This research was aimed to assess the helminth parasites in Schizothorax plagiostomus (the snow trout) from river Panjkora, KP, Pakistan. A total of 88 fish samples (n= 88) were collected from the upper, middle and lower regions of the river through cast nets, hand nets and hooks. All the collected fish samples were examined in the Parasitology Laboratory, Department of Zoology, University of Malakand for helminth parasites during July 2018 to February 2019.Among these fishes nematodes, trematodes, cestodes and acanthocephalan were isolated and identified. Overall prevalence of the fish parasites was 60.22% (53/88). The intensity of the parasite was observed from 1.7% to 4.61%. Highest prevalence was reported in summer season 86.36% while lowest in the winter season 36.36%. This study shows that summer season affects the fish business and result in poor quality fish meat with risk of zoonotic diseases. Adults were highly infected 82.25% while no infection was found in juvenile specimens. Female fish samples had higher prevalence 68.22% than males 48.22%. Fishes of the lower reaches had highest prevalence 60.22% than the upper reaches 40.0%. Fish samples with maximum length and weight were highly 76.92% infected than small sized fish with low body weight. Present study addresses that Rhabdochona schizothoracis in the intestine and Diplozoon paradoxum in gills of snow trout fish has a long term relationship and call as a natural infection in cyprinids and it is zoonotic threat to human.

Resumo O peixe é a principal fonte de proteína de alta qualidade para mais de um bilhão de pessoas no mundo. Contudo, parasitas afetam diretamente a produtividade de peixes e, indiretamente, a saúde humana. Esta pesquisa teve como objetivo avaliar os parasitas helmintos em Schizothorax plagiostomus (truta da neve) do rio Panjkora, Khyber Pakhtunkhwa, Paquistão. Foram coletadas 88 amostras de peixes (n = 88) dos trechos alto, médio e baixo do rio por meio de redes de arrasto, redes de mão e varas de pesca. Todas as amostras de peixes coletadas foram examinadas no Laboratório de Parasitologia do Departamento de Zoologia, da Universidade de Malakand, quanto a parasitas helmintos durante o período de julho de 2018 a fevereiro de 2019. Entre esses peixes, nematódeos, trematódeos, cestóideos e acantocéfalos foram isolados e identificados. A prevalência geral de parasitas de peixes foi de 60,22% (53/88). A intensidade do parasita foi observada de 1,7 a 4,61%. A maior prevalência (86,36%) foi relatada no verão, enquanto a menor (36,36%), no inverno. Este estudo mostra que a temporada de verão afeta o ramo da pesca e resulta em carne de baixa qualidade, com risco de doenças zoonóticas. Os adultos foram altamente infectados (82,25%), enquanto nenhuma infecção foi encontrada em jovens. As amostras de peixes do sexo feminino apresentaram maior prevalência (68,22%) do que as do sexo masculino (48,22%). Os peixes coletados no trecho baixo do rio tiveram maior prevalência (60,22%) do que os do trecho alto (40%). Amostras de peixes com comprimento e peso máximos foram altamente infectadas (76,92%) do que peixes de pequeno porte e baixo peso corporal. O presente estudo aborda que Rhabdochona schizothoracis no intestino e Diplozoon paradoxum em brânquias de peixes de truta da neve possuem uma estreita relação que causa uma infecção natural em ciprinídeos, sendo uma ameaça zoonótica ao ser humano.

Evaluation of ecto and endo parasitic fauna of Schizothorax plagiostomus inhabitants of river Swat, Khyber PakhtunKhwa, Pakistan

Abstract Fish is consumed as a rich and cheaper source of white meat and proteins all over the world. Fish farming is the leading source of income generation throughout the world. The present research study was conducted with aims to investigate the ecto and endo parasitic fauna of S. plagiostomus (snow trout) in River Swat. A total of 96 fish samples were collected on monthly basis from July-2018 to February-2019, from upper, middle and lower reaches. Fish samples were collected with the help of cast net, hand net and fishing rods. Local fishermen and experts help were also sought out for fish collection. 61 out of 96 fishes were found infected by helminth parasites. The total prevalence and intensity of 63.54% and 2.90 was observed respectively during data analysis. Highest monthly prevalence of 83.33% was recorded during July and August, while lowest prevalence of 33.33% was noted during February. Five species of helminths parasites were detected from S. plagiostomus, of which two were trematodes (Rhabdochona shizothoracis and Rhabdochona species), one species of Monogenia (Diplozoon paraddoxum), one species of Cestodes (Nippotaenia species), and one species of Acanthocephalan (Neoechynorhynchus devdevi). Highest parasite wise prevalence, intensity and relative density of 21.87%, 4.09 and 0.895 was noted for R. schizothoracis while the lowest prevalence, intensity and relative density of 4.16%, 1.25 and 0.052 was noted for N. devdevi. Highest infection of 76.08% was observed in adults host while lowest, 40% infestation rate was observed in young fish samples. 73.68% prevalence was observed in female hosts while only 56.90% prevalence was observed in male individuals. Higher infection (83.33%) was noticed during summer season, while lowest infection (44.44%) was observed during winter season. Similarly 71.79% fishes were found infected that were collected from lower reaches of the River Swat than the fish samples collected from upper reaches (52%).

Resumo O peixe é consumido mundialmente por ser uma fonte rica e barata de carne branca e proteínas. A piscicultura é a principal fonte de geração de renda em todo o mundo. O presente estudo foi conduzido com o objetivo de investigar a fauna ecto e endoparasitária de S. plagiostomus (truta da neve) no rio Swat. Foram coletadas mensalmente 96 amostras de peixes, no período de julho de 2018 a fevereiro de 2019, nos trechos alto, médio e baixo com auxílio de redes de arrasto, redes de mão e varas de pesca. Pescadores locais e especialistas também ajudaram na coleta de peixes. Dos 96 peixes encontrados, 61 estavam infectados por parasitas helmintos. A prevalência de 63,54% e a intensidade total de 2,90 foram observadas durante a análise dos dados. A maior prevalência mensal de 83,33% foi registrada nos meses de julho e agosto, enquanto a menor, de 33,33%, em fevereiro. Cinco espécies de parasitas helmintos foram detectadas de S. plagiostomus, dos quais dois eram trematódeos (espécies Rhabdochona shizothoracis e Rhabdochona), uma espécie de monogenia (Diplozoon paradoxum), uma espécie de cestóideos (espécie Nippotaenia) e uma espécie de acantocéfalo (Neoechynorhynchus devdevi). As maiores prevalências de parasitas (21,87%), intensidade (4,09) e densidade relativa (0,895) foram observadas em R. schizothoracis, enquanto as menores prevalências de parasitas (4,16%), intensidade (1,25) e densidade relativa (0,052), em N. devdevi. A infecção mais alta (76,08%) foi constatada em adultos hospedeiros, ao passo que a taxa de infestação mais baixa (40%) foi encontrada em amostras de peixes jovens. A prevalência de 73,68% foi verificada em hospedeiros do sexo feminino, enquanto a de 56,90%, em indivíduos do sexo masculino. Maior infecção (83,33%) foi observada durante a temporada de verão, e a menor infecção (44,44%), durante o inverno. Da mesma forma, 71,79% dos peixes encontrados infectados foram coletados no trecho baixo do rio Swat, enquanto 52%, no trecho alto.

Structural basis of pathogen recognition by an integrated HMA domain in a plant NLR immune receptor.

Plants have evolved intracellular immune receptors to detect pathogen proteins known as effectors. How these immune receptors detect effectors remains poorly understood. Here we describe the structural basis for direct recognition of AVR-Pik, an effector from the rice blast pathogen, by the rice intracellular NLR immune receptor Pik. AVR-PikD binds a dimer of the Pikp-1 HMA integrated domain with nanomolar affinity. The crystal structure of the Pikp-HMA/AVR-PikD complex enabled design of mutations to alter protein interaction in yeast and in vitro, and perturb effector-mediated response both in a rice cultivar containing Pikp and upon expression of AVR-PikD and Pikp in the model plant Nicotiana benthamiana. These data reveal the molecular details of a recognition event, mediated by a novel integrated domain in an NLR, which initiates a plant immune response and resistance to rice blast disease. Such studies underpin novel opportunities for engineering disease resistance to plant pathogens in staple food crops.

Recognition of the Thomsen-Friedenreich pancarcinoma carbohydrate antigen by a lamprey variable lymphocyte receptor.

Variable lymphocyte receptors (VLRs) are leucine-rich repeat proteins that mediate adaptive immunity in jawless vertebrates. VLRs were recently shown to recognize glycans, such as the tumor-associated Thomsen-Friedenreich antigen (TFα; Galß1-3GalNAcα), with a selectivity rivaling or exceeding that of lectins and antibodies. To understand the basis for TFα recognition by one such VLR (VLRB.aGPA.23), we measured thermodynamic parameters for the binding interaction and determined the structure of the VLRB.aGPA.23-TFα complex to 2.2 Å resolution. In the structure, four tryptophan residues form a tight hydrophobic cage encasing the TFα disaccharide that completely excludes buried water molecules. This cage together with hydrogen bonding of sugar hydroxyls to polar side chains explains the exquisite selectivity of VLRB.aGPA.23. The topology of the glycan-binding site of VLRB.aGPA.23 differs markedly from those of lectins or antibodies, which typically consist of long, convex grooves for accommodating the oligosaccharide. Instead, the TFα disaccharide is sandwiched between a variable loop and the concave surface of the VLR formed by the ß-strands of the leucine-rich repeat modules. Longer oligosaccharides are predicted to extend perpendicularly across the ß-strands, requiring them to bend to match the concavity of the VLR solenoid.

Changes in oestrogen receptor alpha expression in the nucleus of the solitary tract of the rat over the oestrous cycle and following ovariectomy.

Oestrogen is capable of modulating autonomic outflow and baroreflex function via actions on groups of neurones in the brainstem. We investigated the presence of oestrogen receptor (ER) alpha in a part of the nucleus of the solitary tract (NTS) associated with central cardiovascular control, aiming to determine whether ERalpha mRNA and protein expression is correlated with levels of circulating oestrogen during the oestrous cycle. Polymerase chain reaction (PCR) detected ERalpha mRNA in the NTS at each stage of the oestrous cycle, from ovariectomised, sham-operated and male rats. Real-time PCR showed variations in ERalpha mRNA expression during the oestrous cycle, with the highest levels seen in oestrus, and lowest levels in metoestrus (P < 0.05 versus oestrus) and proestrus (P < 0.05 versus oestrus). Expression in males was lower than in dioestrus and oestrus females (P < 0.05). After ovariectomy, ERalpha mRNA levels were decreased compared to sham-operated animals (P < 0.01). Confocal fluorescence immunohistochemistry with stereological analysis showed that numbers of ERalpha immunoreactive cell nuclei per mm(3) of tissue in the caudal NTS were significantly greater in proestrus than in other groups of rats (P < 0.05). There were also differences among the groups in the extent of colocalisation of ERalpha in neurones immunoreactive for tyrosine hydroxylase and nitric oxide synthase. These results imply a complex pattern of region-specific oestrogen signalling in the NTS and suggest that ERalpha expression in this important autonomic nucleus may be related to circulating oestrogen levels. This may have consequences for the regulation of autonomic tone and baroreflex sensitivity when oestrogen levels decline, for example following menopause.

Prevalence and chemotherapy of babesiosis among Lohi sheep in the Livestock Experiment Station, Qadirabad, Pakistan, and environs

A total of 310 blood smears were collected from sheep of the Livestock Experiment Station, Qadirabad, Sahiwal district, Pakistan, and surrounding areas. The samples were examined microscopically and 30 (9.67 percent) were positive for babesiosis. The animals were divided into two groups (A and B) for chemotherapy. Group A sheep were treated with diminazene diaceturate while group B animals received imidocarb dipropionate. Drug efficacy was determined by negative blood smear examination. Diminazene diaceturate effectiveness against babesiosis was 80 percent while that of imidocarb dipropionate was 100 percent. Hematological studies revealed a significant decrease in hemoglobin concentrations and hematocrit values for Babesia-positive animals compared to healthy controls.(AU)

Contrasting behavior of conformationally locked carbocyclic nucleosides of adenosine and cytidine as substrates for deaminases.

In addition to the already known differences between adenosine deaminase (ADA) and cytidine deaminase (CDA) in terms of their tertiary structure, the sphere of Zn(+2) coordination, and their reverse stereochemical preference, we present evidence that the enzymes also differ significantly in terms of the North/South conformational preferences for their substrates and the extent to which the lack of the O(4') oxygen affects the kinetics of the enzymatic deamination of carbocyclic substrates. The carbocyclic nucleoside substrates used in this study have either a flexible cyclopentane ring or a rigid bicyclo[3.1.0]hexane scaffold.

Obturator hernia: a diagnostic challenge.

We describe a case of an 82-year-old lady with groin pain secondary to an obturator hernia. She was a diagnostic challenge, and 6 years passed before the obturator hernia was discovered. She presented to hospital with symptoms of bowel obstruction, and a computer tomography (CT) scan of her pelvis revealed an incarcerated obturator hernia. Her surgical management included reinforcement of the obturator foramen with sutures. A recurrence of the obturator hernia 2 years later required an extra-peritoneal mesh repair of the defect in the obturator foramen with a good outcome.

Activation of p16 gene silenced by DNA methylation in cancer cells by phosphoramidate derivatives of 2'-deoxyzebularine.

We report herein the application of the phosphoramidate ProTide technology to improve the metabolism of the DNA methytransferase inhibitor, zebularine (Z). Zebularine is a riboside that must undergo a complex metabolic transformation before reaching the critical 2'-deoxyzebularine 5'-triphosphate (dZTP). Because 2'-deoxyzebularine (dZ) is not phosphorylated and therefore inactive, the ProTide strategy was employed to bypass the lack of phosphorylation of dZ and the inefficient reduction of zebularine 5'-diphosphate by ribonucleotide-diphosphate reductase required for zebularine. Several compounds were identified as more potent inhibitors of DNA methylation and stronger inducers of p16 tumor suppressor gene than zebularine. However, their activity was dependent on the administration of thymidine to overcome the potent inhibition of thymidylate synthase (TS) and deoxycytidine monophosphate (dCMP) deaminase by dZMP, which deprives cells of essential levels of thymidine. Intriguingly, the activity of the ProTides was cell line-dependent, and activation of p16 was manifest only in Cf-Pac-1 pancreatic ductal adenocarcinoma cells.

2'-Deoxy-N4-[2-(4-nitrophenyl)ethoxycarbonyl]-5-azacytidine: a novel inhibitor of DNA methyltransferase that requires activation by human carboxylesterase 1.

2'-Deoxy-N4-[2-(4-nitrophenyl)ethoxycarbonyl]-5-azacytidine (NPEOC-DAC), decitabine with a modification of the N4 position of the azacitidine ring can be used to inhibit DNA methyltransferase. This modification protects the azacitidine ring and can be cleaved by carboxylesterase to release decitabine. NPEOC-DAC was 23-fold less potent at low doses (<10microM) than decitabine at inhibiting DNA methylation, and was also associated with a 3-day delay in its effect. However, at doses > or = 10microM NPEOC-DAC was more effective at inhibiting DNA methylation. Theses differences between decitabine and NPEOC-DAC are dependent on the cleavage of the carboxylester bond, and could be potentially exploited pharmacologically.

The triphosphate of beta-D-4'-C-ethynyl-2',3'-dideoxycytidine is the preferred enantiomer substrate for HIV reverse transcriptase.

The enantioselective synthesis of the beta-d (1) enantiomer of 4'-C-ethynyl-2',3'-dideoxycytidine confirms an earlier stereochemical assignment that was strictly based on the ability of HIV reverse transcriptase and its M184V mutant to discriminate between the d- and l-configuration of nucleoside 5'-triphosphates.

Phase I pharmacokinetic studies evaluating single and multiple doses of oral GW572016, a dual EGFR-ErbB2 inhibitor, in healthy subjects.

GW572016 is a dual EGFR-ErbB2 inhibitor that has promise as an anticancer agent. Two phase I studies were conducted to determine the safety, tolerability and pharmacokinetics of single and multiple doses given to healthy subjects. The single dose study evaluated two groups of eight subjects in an ascending dose, 4-way cross-over, while the multiple dose study evaluated twenty-seven healthy volunteers in an ascending dose, double-blind, randomized, placebo-controlled, staggered parallel design. No serious adverse events were seen in either study. The most common adverse events for subjects receiving GW572016 were headache, diarrhea, rash, cold symptoms, gastrointestinal symptoms, and elevated LFTs, which were similar between treatment and placebo groups. Absorption of single doses of GW572016 was slightly delayed, with median t(lag) of 15 minutes (range 0-90 minutes) and achieved peak serum concentrations at a median of three hours (range 1.5-6 hours) post-dose. Serum concentrations after multiple doses of GW572016 demonstrated no significant accumulation at the 25 mg dose, and approximately 50% accumulation at the 100 mg and 175 mg doses, achieving steady state in six to seven days. A modest time-dependent increase in serum concentrations also was detected with multiple doses of GW572016. Single and multiple oral doses of GW572016 were well tolerated in healthy subjects, and resulted in dose-related systemic exposure of GW572016.

An evaluation of the beta-1 adrenergic receptor Arg389Gly polymorphism in individuals at risk of coronary events. A WOSCOPS substudy.

AIMS: The Glycine389 variant of the beta-1 adrenergic receptor generates markedly less cAMP when stimulated in vitro than the more prevalent Arginine389 variant and may confer protection against coronary events similar to that observed with beta-blockers. The aim of this study was to ascertain whether this Glycine389 variant protects against coronary events. METHODS AND RESULTS: We identified the genotype at position 389 of the beta1AR in 1554 individuals taken from men enrolled in the West of Scotland Coronary Prevention Study. Men with a coronary event (event group) were each matched for age and smoking status with two control subjects from the same cohort who had not had a coronary event (control group). We compared the distribution of genotypes in the event and control groups. Conditional logistic regression was used to calculate odds ratios for each of the genotypes. The prevalence of the three genotypes in the entire cohort was ArgArg 53.5%, ArgGly 39.6%, GlyGly 6.9%. The Arg389Gly beta-1 adrenergic receptor polymorphism was not associated with coronary events. Using the ArgArg genotype as the reference, the odds ratio for the ArgGly genotype was 1.1 (95% CI, 0.88-1.38) and for the GlyGly genotype it was 1.05 (95% CI, 0.68-1.62). CONCLUSION: Our longitudinal case-control study demonstrates that the Glycine389 variant of the beta-1 adrenergic receptor does not protect against coronary events.

In vivo GM-CSF prevents dexamethasone suppression of killing of Aspergillus fumigatus conidia by bronchoalveolar macrophages.

Dexamethasone (DEX) is a potent immunosuppressive agent used in the treatment of several disorders. However, despite its beneficial effects, DEX puts patients at risk for opportunistic infections, especially pulmonary aspergillosis. Previously we reported that in vitro granulocyte-macrophage colony-stimulating factor (GM-CSF) blocks the immunosuppressive action of DEX on bronchoalveolar macrophages (BAMs). Here we report that BAMs freshly isolated from mice treated intraperitoneally with DEX for 24 h had significantly (P<0.01) reduced killing of conidia, i.e., 15 +/- 5% conidia killed by BAMs from DEX-treated mice versus 35 +/- 3% by BAMs from mice given saline, 38 +/- 5% by BAMs from mice given GM-CSF, and 39 +/- 1% by BAMs from mice given both DEX and GM-CSF. On the other hand, in another compartment GM-CSF could not block the DEX reduction of spleen weight and spleen cellularity. Unlike GM-CSF, granulocyte colony-stimulating factor did not block DEX suppression of BAMs. GM-CSF given 24 h before DEX resulted in blocking of DEX suppression of BAM conidiacidal activity. However, when DEX was given 24 h before GM-CSF, DEX suppression of BAM was not reversed. These data show that GM-CSF in vivo blocks the in vivo immunosuppressive effects of DEX on BAM killing of conidia and suggest a potential use of GM-CSF in patients at risk for aspergillosis due to immunosuppressive DEX treatment.

Protection of bronchoalveolar macrophages by granulocyte-macrophage colony-stimulating factor against dexamethasone suppression of fungicidal activity for Aspergillus fumigatus conidia.

The objectives of this study were to: (i) see if granulocyte macrophage colony-stimulating factor (GM-CSF) could protect bronchoalveolar macrophages (BAM) against suppression by dexamethasone (DEX) and (ii) test the combined effect of GM-CSF and DEX on lymphocyte responses. Murine BAM killed Aspergillus fumigatus conidia by 33 +/- 4% (mean +/- SD) in a 2.5-h assay, unaffected by GM-CSF treatment. Killing by BAM treated with DEX (10(-7) M) for 48 h in vitro was reduced to 13 +/- 6%; however, if GM-CSF (500 U ml(-1)) was present during DEX treatment of BAM, killing of conidia (33 +/- 2%) by BAM was preserved. By contrast, DEX suppression of lymphocyte responses to concanavalin A was maintained during co-culture with GM-CSF. In sequence treatment experiments, initial treatment of BAM with GM-CSF protected against subsequent treatment with DEX. When macrophages were pretreated with DEX, GM-CSF could reverse suppression even when added subsequently, provided DEX treatment was discontinued. These data suggest that it may be possible to suppress lymphocyte responses with DEX, yet at the same time maintain BAM defenses with GM-CSF against pulmonary infections by conidia of A. fumigatus.