Assessment of proline function in higher plants under extreme temperatures.

Climate change and abiotic stress factors are key players in crop losses worldwide. Among which, extreme temperatures (heat and cold) disturb plant growth and development, reduce productivity and, in severe cases, lead to plant death. Plants have developed numerous strategies to mitigate the detrimental impact of temperature stress. Exposure to stress leads to the accumulation of various metabolites, e.g. sugars, sugar alcohols, organic acids and amino acids. Plants accumulate the amino acid 'proline' in response to several abiotic stresses, including temperature stress. Proline abundance may result from de novo synthesis, hydrolysis of proteins, reduced utilization or degradation. Proline also leads to stress tolerance by maintaining the osmotic balance (still controversial), cell turgidity and indirectly modulating metabolism of reactive oxygen species. Furthermore, the crosstalk of proline with other osmoprotectants and signalling molecules, e.g. glycine betaine, abscisic acid, nitric oxide, hydrogen sulfide, soluble sugars, helps to strengthen protective mechanisms in stressful environments. Development of less temperature-responsive cultivars can be achieved by manipulating the biosynthesis of proline through genetic engineering. This review presents an overview of plant responses to extreme temperatures and an outline of proline metabolism under such temperatures. The exogenous application of proline as a protective molecule under extreme temperatures is also presented. Proline crosstalk and interaction with other molecules is also discussed. Finally, the potential of genetic engineering of proline-related genes is explained to develop 'temperature-smart' plants. In short, exogenous application of proline and genetic engineering of proline genes promise ways forward for developing 'temperature-smart' future crop plants.

Evaluation of economic loss caused by Indian crested porcupine (Hystrix indica) in agricultural land of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan / Avaliação da perda econômica causada pelo porco-espinho de crista indiano (Hystrix indica) em terras agrícolas dos distritos de Muzaffarabad, Azad Jammu e Caxemira, Paquistão

Abstract The Indian crested porcupine (Hystrix indica) is a vertebrate pest of agricultural lands and forest. The study was aimed to report the damage to local crops by the Indian crested porcupine (Hystrix indica) in the Muzaffarabad District. A survey was conducted to identify the porcupine-affected areas and assess the crop damage to the local farmers in district Muzaffarabad Azad Jammu and Kashmir (AJK) from May 2017 to October 2017. Around 19 villages were surveyed, and a sum of 191 semi-structured questionnaires was distributed among farmers. Crop damage was found highest in village Dhanni where a porcupine destroyed 175 Kg/Kanal of the crops. Regarding the total magnitude of crop loss, village Danna and Koomi kot were the most affected areas. More than half (51.8%) of the respondents in the study area suffered the economic loss within the range of 101-200$, and (29.8%) of the people suffered losses in the range of 201-300$ annually. Among all crops, maize (Zea mays) was found to be the most damaged crop ranging between 1-300 Kg annually. In the study area, porcupine also inflicted a lot of damages to some important vegetables, including spinach (Spinacia oleracea), potato (Solanum tuberosum) and onion (Allium cepa). It was estimated that, on average, 511Kg of vegetables are destroyed by porcupine every year in the agricultural land of Muzaffarabad. It was concluded that the Indian crested porcupine has a devastating effect on agriculture which is an important source of income and food for the local community. Developing an effective pest control strategy with the help of the local government and the Wildlife department could help the farmers to overcome this problem.

Resumo O porco-espinho de crista indiano (Hystrix indica) é uma praga vertebrada de terras agrícolas e florestais. No estudo atual, o dano às plantações locais pelo porco-espinho de crista indiano (Hystrix indica) foi relatado pela primeira vez no distrito de Muzaffarabad. O estudo foi projetado para investigar a perda econômica causada pelo porco-espinho de crista indiano (Hystrix indica) nos distritos de Muzaffarabad, Azad Jammu e Caxemira (AJK) de maio de 2017 a outubro de 2017. Um estudo baseado em pesquisa foi conduzido para identificar as áreas afetadas por porcos-espinhos e avaliar os danos às colheitas para os agricultores locais. Cerca de 19 aldeias foram pesquisadas e um total de 191 questionários semiestruturados foi distribuído entre os agricultores. Os danos às colheitas foram encontrados mais intensamente na aldeia Dhanni, onde um porco-espinho destruiu 175 kg / Kanal das colheitas. Em relação à magnitude total da perda de safra, as aldeias Danna e Koomi Kot foram as áreas mais afetadas. Mais da metade (51,8%) dos entrevistados na área de estudo sofreu perdas econômicas na faixa de 101-200 $, e 29,8% das pessoas sofreram perdas na faixa de 201-300 $ anualmente. Entre todas as culturas, o milho (Zea mays) foi considerado a cultura mais danificada, variando entre 1-300 kg anualmente. Na área de estudo, o porco-espinho também causou muitos danos a alguns vegetais importantes, incluindo espinafre (Spinacia oleracea), batata (Solanum tuberosum) e cebola (Allium cepa). Estimou-se que, em média, 511 kg de vegetais são destruídos pelo porco-espinho todos os anos nas terras agrícolas de Muzaffarabad. Concluiu-se que o porco-espinho de crista indiano tem um efeito devastador na agricultura, que é importante fonte de renda e alimento para a comunidade local. O desenvolvimento de uma estratégia eficaz de controle de pragas com a ajuda do governo local e do Departamento de Vida Selvagem pode ajudar os agricultores a superar esse problema.

Evaluation of economic loss caused by Indian crested porcupine (Hystrix indica) in agricultural land of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan / Avaliação da perda econômica causada pelo porco-espinho de crista indiano (Hystrix indica) em terras agrícolas dos distritos de Muzaffarabad, Azad Jammu e Caxemira, Paquistão

The Indian crested porcupine (Hystrix indica) is a vertebrate pest of agricultural lands and forest. The study was aimed to report the damage to local crops by the Indian crested porcupine (Hystrix indica) in the Muzaffarabad District. A survey was conducted to identify the porcupine-affected areas and assess the crop damage to the local farmers in district Muzaffarabad Azad Jammu and Kashmir (AJK) from May 2017 to October 2017. Around 19 villages were surveyed, and a sum of 191 semi-structured questionnaires was distributed among farmers. Crop damage was found highest in village Dhanni where a porcupine destroyed 175 Kg/Kanal of the crops. Regarding the total magnitude of crop loss, village Danna and Koomi kot were the most affected areas. More than half (51.8%) of the respondents in the study area suffered the economic loss within the range of 101-200$, and (29.8%) of the people suffered losses in the range of 201-300$ annually. Among all crops, maize (Zea mays) was found to be the most damaged crop ranging between 1-300 Kg annually. In the study area, porcupine also inflicted a lot of damages to some important vegetables, including spinach (Spinacia oleracea), potato (Solanum tuberosum) and onion (Allium cepa). It was estimated that, on average, 511Kg of vegetables are destroyed by porcupine every year in the agricultural land of Muzaffarabad. It was concluded that the Indian crested porcupine has a devastating effect on agriculture which is an important source of income and food for the local community. Developing an effective pest control strategy with the help of the local government and the Wildlife department could help the farmers to overcome this problem.

O porco-espinho de crista indiano (Hystrix indica) é uma praga vertebrada de terras agrícolas e florestais. No estudo atual, o dano às plantações locais pelo porco-espinho de crista indiano (Hystrix indica) foi relatado pela primeira vez no distrito de Muzaffarabad. O estudo foi projetado para investigar a perda econômica causada pelo porco espinho de crista indiano (Hystrix indica) nos distritos de Muzaffarabad, Azad Jammu e Caxemira (AJK) de maio de 2017 a outubro de 2017. Um estudo baseado em pesquisa foi conduzido para identificar as áreas afetadas por porcos-espinhos e avaliar os danos às colheitas para os agricultores locais. Cerca de 19 aldeias foram pesquisadas e um total de 191 questionários semiestruturados foi distribuído entre os agricultores. Os danos às colheitas foram encontrados mais intensamente na aldeia Dhanni, onde um porco-espinho destruiu 175 kg / Kanal das colheitas. Em relação à magnitude total da perda de safra, as aldeias Danna e Koomi Kot foram as áreas mais afetadas. Mais da metade (51,8%) dos entrevistados na área de estudo sofreu perdas econômicas na faixa de 101-200 $, e 29,8% das pessoas sofreram perdas na faixa de 201-300 $ anualmente. Entre todas as culturas, o milho (Zea mays) foi considerado a cultura mais danificada, variando entre 1-300 kg anualmente. Na área de estudo, o porco-espinho também causou muitos danos a alguns vegetais importantes, incluindo espinafre (Spinacia oleracea), batata (Solanum tuberosum) e cebola (Allium cepa). Estimou-se que, em média, 511 kg de vegetais são destruídos pelo porco-espinho todos os anos nas terras agrícolas de Muzaffarabad. Concluiu-se que o porco-espinho de crista indiano tem um efeito devastador na agricultura, que é importante fonte de renda e alimento para a comunidade local. O desenvolvimento de uma estratégia eficaz de controle de pragas com a ajuda do governo local e do Departamento de Vida Selvagem pode ajudar os agricultores a superar esse problema.

Evaluation of economic loss caused by Indian crested porcupine (Hystrix indica) in agricultural land of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan

Abstract The Indian crested porcupine (Hystrix indica) is a vertebrate pest of agricultural lands and forest. The study was aimed to report the damage to local crops by the Indian crested porcupine (Hystrix indica) in the Muzaffarabad District. A survey was conducted to identify the porcupine-affected areas and assess the crop damage to the local farmers in district Muzaffarabad Azad Jammu and Kashmir (AJK) from May 2017 to October 2017. Around 19 villages were surveyed, and a sum of 191 semi-structured questionnaires was distributed among farmers. Crop damage was found highest in village Dhanni where a porcupine destroyed 175 Kg/Kanal of the crops. Regarding the total magnitude of crop loss, village Danna and Koomi kot were the most affected areas. More than half (51.8%) of the respondents in the study area suffered the economic loss within the range of 101-200$, and (29.8%) of the people suffered losses in the range of 201-300$ annually. Among all crops, maize (Zea mays) was found to be the most damaged crop ranging between 1-300 Kg annually. In the study area, porcupine also inflicted a lot of damages to some important vegetables, including spinach (Spinacia oleracea), potato (Solanum tuberosum) and onion (Allium cepa). It was estimated that, on average, 511Kg of vegetables are destroyed by porcupine every year in the agricultural land of Muzaffarabad. It was concluded that the Indian crested porcupine has a devastating effect on agriculture which is an important source of income and food for the local community. Developing an effective pest control strategy with the help of the local government and the Wildlife department could help the farmers to overcome this problem.

Resumo O porco-espinho de crista indiano (Hystrix indica) é uma praga vertebrada de terras agrícolas e florestais. No estudo atual, o dano às plantações locais pelo porco-espinho de crista indiano (Hystrix indica) foi relatado pela primeira vez no distrito de Muzaffarabad. O estudo foi projetado para investigar a perda econômica causada pelo porco-espinho de crista indiano (Hystrix indica) nos distritos de Muzaffarabad, Azad Jammu e Caxemira (AJK) de maio de 2017 a outubro de 2017. Um estudo baseado em pesquisa foi conduzido para identificar as áreas afetadas por porcos-espinhos e avaliar os danos às colheitas para os agricultores locais. Cerca de 19 aldeias foram pesquisadas e um total de 191 questionários semiestruturados foi distribuído entre os agricultores. Os danos às colheitas foram encontrados mais intensamente na aldeia Dhanni, onde um porco-espinho destruiu 175 kg / Kanal das colheitas. Em relação à magnitude total da perda de safra, as aldeias Danna e Koomi Kot foram as áreas mais afetadas. Mais da metade (51,8%) dos entrevistados na área de estudo sofreu perdas econômicas na faixa de 101-200 $, e 29,8% das pessoas sofreram perdas na faixa de 201-300 $ anualmente. Entre todas as culturas, o milho (Zea mays) foi considerado a cultura mais danificada, variando entre 1-300 kg anualmente. Na área de estudo, o porco-espinho também causou muitos danos a alguns vegetais importantes, incluindo espinafre (Spinacia oleracea), batata (Solanum tuberosum) e cebola (Allium cepa). Estimou-se que, em média, 511 kg de vegetais são destruídos pelo porco-espinho todos os anos nas terras agrícolas de Muzaffarabad. Concluiu-se que o porco-espinho de crista indiano tem um efeito devastador na agricultura, que é importante fonte de renda e alimento para a comunidade local. O desenvolvimento de uma estratégia eficaz de controle de pragas com a ajuda do governo local e do Departamento de Vida Selvagem pode ajudar os agricultores a superar esse problema.

Ecological impacts of climate change on the snow leopard (Panthera unica) in South Asia / Impactos ecológicos da mudança climática no leopardo-da-neve (Panthera uncia) no sul da Ásia

Snow leopard (Panthera unica) is a felid which lives in the highly rugged areas of alpine regions in different mountain ranges of South and Central Asia. This solitary animal needs large spaces for its ranges but due to climate change and relatively faster rate of global warming in South Asian mountain ranges, its habitat is going to shrink and fragment by tree-line shifts and change in hydrology of the area. Vegetative modification of montane flora and competition with domestic goats will create its prey's population to decline along with a chance of a direct conflict and competition with the common leopard. Common leopard being more adaptable, grouped, and larger in size can be a significant stressor for a smaller and solitary snow leopard. Habitat would shrink, and snow leopard can possibly move upslope or northward to central Asian ranges and their predicted migratory patterns are unknown.

O leopardo-das-neves (Panthera uncia) é um felídeo que vive nas áreas altamente acidentadas das regiões alpinas, em diferentes cadeias de montanhas do sul e do centro da Ásia. A revisão atual tem o objetivo de abordar a questão da possível perda de hábitat do leopardo da neve em razão da interferência humana nas montanhas do sul e do centro da Ásia. Esse animal solitário precisa de grandes espaços para suas faixas, mas, devido às mudanças climáticas e à taxa relativamente mais rápida de aquecimento global nas cadeias montanhosas do sul da Ásia, seu hábitat vai encolher e fragmentar-se por mudanças nas linhas de árvores e na hidrologia da área. Este artigo teve o objetivo de rever vários estudos sobre os impactos das mudanças climáticas em um futuro próximo e distante sobre os hábitats do leopardo-da-neve. O estudo revelou que a modificação vegetativa da flora da montanha e a competição com cabras domésticas poderiam ocasionar na diminuição da população de suas presas, com a chance de um conflito direto e competição com leopardo comum. Leopardo comum sendo mais adaptável, agrupado e maior em tamanho pode ser um estressor significativo para um leopardo-da-neve menor e solitário. Conclusão: devido ao encolhimento do hábitat, o leopardo-da-neve pode, possivelmente, mover-se para cima ou para o norte, para faixas da Ásia Central com padrões migratórios não previstos.

Ecological impacts of climate change on the snow leopard (Panthera unica) in South Asia / Impactos ecológicos da mudança climática no leopardo-da-neve (Panthera uncia) no sul da Ásia

Snow leopard (Panthera unica) is a felid which lives in the highly rugged areas of alpine regions in different mountain ranges of South and Central Asia. This solitary animal needs large spaces for its ranges but due to climate change and relatively faster rate of global warming in South Asian mountain ranges, its habitat is going to shrink and fragment by tree-line shifts and change in hydrology of the area. Vegetative modification of montane flora and competition with domestic goats will create its prey's population to decline along with a chance of a direct conflict and competition with the common leopard. Common leopard being more adaptable, grouped, and larger in size can be a significant stressor for a smaller and solitary snow leopard. Habitat would shrink, and snow leopard can possibly move upslope or northward to central Asian ranges and their predicted migratory patterns are unknown.

O leopardo-das-neves (Panthera uncia) é um felídeo que vive nas áreas altamente acidentadas das regiões alpinas, em diferentes cadeias de montanhas do sul e do centro da Ásia. A revisão atual tem o objetivo de abordar a questão da possível perda de hábitat do leopardo da neve em razão da interferência humana nas montanhas do sul e do centro da Ásia. Esse animal solitário precisa de grandes espaços para suas faixas, mas, devido às mudanças climáticas e à taxa relativamente mais rápida de aquecimento global nas cadeias montanhosas do sul da Ásia, seu hábitat vai encolher e fragmentar-se por mudanças nas linhas de árvores e na hidrologia da área. Este artigo teve o objetivo de rever vários estudos sobre os impactos das mudanças climáticas em um futuro próximo e distante sobre os hábitats do leopardo-da-neve. O estudo revelou que a modificação vegetativa da flora da montanha e a competição com cabras domésticas poderiam ocasionar na diminuição da população de suas presas, com a chance de um conflito direto e competição com leopardo comum. Leopardo comum sendo mais adaptável, agrupado e maior em tamanho pode ser um estressor significativo para um leopardo-da-neve menor e solitário. Conclusão: devido ao encolhimento do hábitat, o leopardo-da-neve pode, possivelmente, mover-se para cima ou para o norte, para faixas da Ásia Central com padrões migratórios não previstos.

Ecological impacts of climate change on the snow leopard (Panthera unica) in South Asia

Abstract Snow leopard (Panthera unica) is a felid which lives in the highly rugged areas of alpine regions in different mountain ranges of South and Central Asia. This solitary animal needs large spaces for its ranges but due to climate change and relatively faster rate of global warming in South Asian mountain ranges, its habitat is going to shrink and fragment by tree-line shifts and change in hydrology of the area. Vegetative modification of montane flora and competition with domestic goats will create its preys population to decline along with a chance of a direct conflict and competition with the common leopard. Common leopard being more adaptable, grouped, and larger in size can be a significant stressor for a smaller and solitary snow leopard. Habitat would shrink, and snow leopard can possibly move upslope or northward to central Asian ranges and their predicted migratory patterns are unknown.

Resumo O leopardo-das-neves (Panthera uncia) é um felídeo que vive nas áreas altamente acidentadas das regiões alpinas, em diferentes cadeias de montanhas do sul e do centro da Ásia. A revisão atual tem o objetivo de abordar a questão da possível perda de hábitat do leopardo da neve em razão da interferência humana nas montanhas do sul e do centro da Ásia. Esse animal solitário precisa de grandes espaços para suas faixas, mas, devido às mudanças climáticas e à taxa relativamente mais rápida de aquecimento global nas cadeias montanhosas do sul da Ásia, seu hábitat vai encolher e fragmentar-se por mudanças nas linhas de árvores e na hidrologia da área. Este artigo teve o objetivo de rever vários estudos sobre os impactos das mudanças climáticas em um futuro próximo e distante sobre os hábitats do leopardo-da-neve. O estudo revelou que a modificação vegetativa da flora da montanha e a competição com cabras domésticas poderiam ocasionar na diminuição da população de suas presas, com a chance de um conflito direto e competição com leopardo comum. Leopardo comum sendo mais adaptável, agrupado e maior em tamanho pode ser um estressor significativo para um leopardo-da-neve menor e solitário. Conclusão: devido ao encolhimento do hábitat, o leopardo-da-neve pode, possivelmente, mover-se para cima ou para o norte, para faixas da Ásia Central com padrões migratórios não previstos.

Evaluation of economic loss caused by Indian crested porcupine (Hystrix indica) in agricultural land of district Muzaffarabad, Azad Jammu and Kashmir, Pakistan.

The Indian crested porcupine (Hystrix indica) is a vertebrate pest of agricultural lands and forest. The study was aimed to report the damage to local crops by the Indian crested porcupine (Hystrix indica) in the Muzaffarabad District. A survey was conducted to identify the porcupine-affected areas and assess the crop damage to the local farmers in district Muzaffarabad Azad Jammu and Kashmir (AJK) from May 2017 to October 2017. Around 19 villages were surveyed, and a sum of 191 semi-structured questionnaires was distributed among farmers. Crop damage was found highest in village Dhanni where a porcupine destroyed 175 Kg/Kanal of the crops. Regarding the total magnitude of crop loss, village Danna and Koomi kot were the most affected areas. More than half (51.8%) of the respondents in the study area suffered the economic loss within the range of 101-200$, and (29.8%) of the people suffered losses in the range of 201-300$ annually. Among all crops, maize (Zea mays) was found to be the most damaged crop ranging between 1-300 Kg annually. In the study area, porcupine also inflicted a lot of damages to some important vegetables, including spinach (Spinacia oleracea), potato (Solanum tuberosum) and onion (Allium cepa). It was estimated that, on average, 511Kg of vegetables are destroyed by porcupine every year in the agricultural land of Muzaffarabad. It was concluded that the Indian crested porcupine has a devastating effect on agriculture which is an important source of income and food for the local community. Developing an effective pest control strategy with the help of the local government and the Wildlife department could help the farmers to overcome this problem.

Ecological impacts of climate change on the snow leopard (Panthera unica) in South Asia.

Snow leopard (Panthera unica) is a felid which lives in the highly rugged areas of alpine regions in different mountain ranges of South and Central Asia. This solitary animal needs large spaces for its ranges but due to climate change and relatively faster rate of global warming in South Asian mountain ranges, its habitat is going to shrink and fragment by tree-line shifts and change in hydrology of the area. Vegetative modification of montane flora and competition with domestic goats will create its prey's population to decline along with a chance of a direct conflict and competition with the common leopard. Common leopard being more adaptable, grouped, and larger in size can be a significant stressor for a smaller and solitary snow leopard. Habitat would shrink, and snow leopard can possibly move upslope or northward to central Asian ranges and their predicted migratory patterns are unknown.

A study on the prevalence of a tick-transmitted pathogen, Theileria annulata, and hematological profile of cattle from Southern Punjab (Pakistan).

The present study was carried out to determine the prevalence of Theileria annulata in large ruminants in Southern Punjab (Pakistan). Blood samples were collected from 144 large ruminants, consisting of 105 cattle and 39 buffaloes, from six districts of Southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar, and Vehari. Data on the characteristics of the animals and herds were collected through questionnaires. The age of animals (P = 0.02), presence of ticks on animals (P = 0.02), and presence of ticks on dogs associated with herds (P = 0.05) were among the major risk factors involved in the spread of tropical theileriosis in the study area. Two different parasite detection techniques, PCR amplification and screening of Giemsa-stained slides, were compared, and it was found that PCR amplification is a more sensitive tool (19% parasite detection) as compared to smear scanning (3% parasite detection) for the detection of T. annulata. Twenty eight out of 144 animals produced the 721-bp fragment specific for T. annulata from five out of six sampling districts. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite-positive and parasite-negative samples to assess the effect of T. annulata on the blood and serological profile of infected animals.

Possible molecular mechanisms of cryoablation-induced impotence in a rat model.

OBJECTIVES: Cryoablation of the prostate has been reported to induce impotence as a consequence of cavernosal nerve injury. This study is designed to investigate the early and late effects of cavernosal nerve cryoablation on growth factor expression and erectile function in a rat model. METHODS: Forty male rats were divided into two groups (n=20 each). The first group underwent unilateral cavernosal nerve freezing (experimental group). Before their euthanization at 1 and 3 months (10 rats each), erectile function was assessed by electrostimulation of the cavernous nerves. The second group served as the control and was killed at the same time points. Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) techniques were used to identify protein and gene expression of nerve growth factor (NGF), transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), and insulin-like growth factor-1 (IGF-1) in the rat penis and pelvic ganglia. RESULTS: Electrostimulation of the frozen nerve after 3 months revealed a significantly higher maximal intracavernosal pressure and a shorter latency period than in the 1-month group. At 3 months, immunoblot showed upregulation of NGF, TGF-alpha, and the precursor form of IGF-1 protein expression in the penile tissue; RT-PCR showed downregulation of NGF gene expression in the pelvic ganglia of the frozen side. CONCLUSIONS: The results show that erectile function decreased at 1 month and then partially recovered 3 months after cavernosal nerve freezing. This alteration in erectile function was associated with differential gene and protein expression of the growth factors (NGF, TGF-alpha, EGF, and IGF-1). Further studies are required to elucidate the potential role of these growth factors in the prevention and treatment of cryoablation-induced impotence.

Effect of cavernous nerve freezing on protein and gene expression of nitric oxide synthase in the rat penis and pelvic ganglia.

PURPOSE: Cryoablation of the prostate has been reported to induce impotence as a result of cavernous nerve injury. This study is designed to investigate the early and late effects of cavernosal nerve cryoablation on erectile function and nitric oxide synthase (NOS) containing nerves in the rat penis and pelvic ganglia. MATERIALS AND METHODS: Forty male rats were divided into two groups (n = 20 each). The first group underwent unilateral cavernous nerve freezing (experimental group). Before their euthanization at 1 and 3 months (10 rats each), erectile function was assessed by electrostimulation of the cavernous nerves. The second group served as a control and was euthanized at the same time points. Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining was used to identify NOS in the penile nerve fibers of the mid-shaft segment and neurons of the pelvic ganglia. Western blot, RT-PCR and immunostaining techniques were used to identify protein and gene expression of eNOS, iNOS, and nNOS. RESULTS: One month after unilateral cavernosal nerve freezing, there was a significant decrease in NOS-containing nerve fibers in the dorsal and intracavernosal nerves and ipsilateral pelvic ganglia as compared with the intact side. At three months, the number of NOS containing nerve fibers in the above-mentioned nerves and ganglia showed a significant increase on the frozen side but not the intact side. Electrostimulation of the frozen nerve after three months revealed a significantly higher maximal intracavernosal pressure and a shorter latency period than the one-month group. At three months, immunoblot showed up-regulation of eNOS and nNOS protein expression in the penile tissue. RT-PCR showed up-regulation of nNOS gene expression in the pelvic ganglia of the frozen side. Immunostaining confirmed the results of western blot and showed significant increase of the nNOS positive staining in the frozen side of the penile tissue after three months. There was no difference in iNOS after three months between both sides. Our repeated eNOS immunostaining was not successful. CONCLUSIONS: The results show that intracavernous pressure response to neurostimulation markedly decreased at one month and then partially recovered three months after cavernosal nerve freezing. A similar pattern of changes of the NOS-containing nerve fibers in dorsal nerves, intracavernosal nerves, and pelvic ganglia were observed. This alteration in erectile function and NOS-containing nerves was associated with differential gene and protein expression of the three subtypes of NOS.

Histological and ultrastructural alterations in an animal model of Peyronie's disease.

OBJECTIVE: To determine the role of transforming growth factor beta (TGF-beta), one of the cytokines known to induce tissue fibrosis, in the induction of a Peyronie's-like condition, and to produce an animal model for the further study of Peyronie's disease. MATERIALS AND METHODS: Twenty-four adult male Sprague-Dawley rats were divided into two groups: in group 1, different concentrations of cytomodulin, a synthetic heptapeptide with TGF-beta-like activity, were injected into the tunica of each of 18 rats and six rats group 2 received saline injections as a control. The tunical tissues were taken after 3 days, 2 and 6 weeks and were examined histologically using Hart and trichrome stains. Electron microscopy was used to examine the ultrastructural changes in the same tissue samples. RESULTS: There were histological and ultrastructural alterations in 15 of 18 rats in group 1 (cytomodulin-injected), especially in tissue examined after 6 weeks. The most prominent histological changes were chronic inflammatory cellular infiltration, focal and diffuse elastosis, thickening, disorganization and clumping of the collagen bundles. The ultrastructural changes were in the form of densely packed collagen, fragmented and scarce elastic fibres, separation of neuronal fibres by interposing clumps of packed collagen, and perivascular collagen deposition as a part of the reorganization of the interstitial matrix. CONCLUSION: Cytomodulin can induce a Peyronie's-like condition in the rat penis, which may explain the role of TGF-beta in the pathogenesis of Peyronie's disease. With further refinement, such rats may be used as an experimental model for studies of Peyronie's disease.