RESUMO
The oviduct plays a crucial role in fertilization, gamete maturation and embryo transport. Prostaglandins are some of the main factors determining its roles. The present study investigated the influence of oestrus synchronization and superovulation on prostaglandins synthesis in the porcine oviduct. Mature cross-bred gilts after exhibiting oestrous cycles were divided into four groups: I, cyclic; II, inseminated; III, synchronized and inseminated; and IV, superovulated and inseminated. Oviducts were collected on the third day of the oestrous cycle or after insemination and divided into isthmus and ampullary parts. This study demonstrated lower mRNA (in the isthmus and ampulla; p < 0.05, p < 0.001, respectively) and protein prostaglandin endoperoxide synthase 2 expression (in the isthmus; p < 0.001) in gilts treated with human chorionic gonadotrophin/equine chorionic gonadotrophin (hCG/eCG) compared with Group II that were inseminated only. In addition, hCG and eCG treatment decreased mPGES-1 mRNA levels in Groups III and IV, in both the isthmus (p < 0.01 in III, p < 0.001 in IV) and ampulla (p < 0.001). The prostaglandin E2 content of oviductal tissues was significantly lower in Groups III (p < 0.05) and IV (p < 0.01 in isthmus, p < 0.0001 in ampulla) compared with Group II. mRNA and protein levels of PGFS in Group IV in the oviductal isthmus were higher (p < 0.01) compared with the non-treated Group II. In effect, the amount of prostaglandin F2α in oviductal tissues of gilts treated with hCG/eCG was significantly elevated (p < 0.001 in isthmus of Groups III and IV; p < 0.05 in ampulla of Group IV). Differential expression of the factors analysed in gilts treated with exogenous gonadotrophins suggests that hCG/eCG stimulation affects prostaglandins synthesis pathway. These changes can alter oviduct functions and in turn affect gamete maturation and fertilization as well as development of embryos and their transport to the uterus.
Assuntos
Gonadotropina Coriônica/farmacologia , Tubas Uterinas/efeitos dos fármacos , Tubas Uterinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Prostaglandinas/metabolismo , Suínos/fisiologia , Animais , Feminino , Cavalos , Humanos , Prostaglandinas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Establishment of pregnancy in pigs requires continuous function of corpora lutea and endometrial preparation for embryo implantation. Progesterone regulates expression of many proteins necessary for endometrial remodelling and embryo-maternal communications. Attaining the uterine receptivity involves progesterone priming and loss of progesterone receptors in the uterine epithelium before days 10-12 after oestrus. Spermatozoa and oocytes in oviduct alter secretion of specific proteins that exert beneficial effect on gametes and embryos. Moreover, an appropriate leucocyte activation and maintenance of delicate cytokine balance within the oviduct and uterus are important for early pregnancy. This early local immune response is rather mediated by seminal plasma components. These components also influence prostaglandin (PG) synthesis in the oviduct that is important for gamete and embryo transport. Pregnancy establishment requires the biphasic pattern of oestrogen secretion by conceptuses on days 11-12 and 15-30. Conceptus affects lipid signalling system consisting of prostaglandins and lysophosphatic acid. PG synthesis is changed by conceptus signals in favour of luteoprotective PGE(2) . Additionally, existence of PGE(2) positive feedback loop in the endometrium contributes to increased PGE(2) /PGF(2α) ratio during the peri-implantation period. PGE(2) through endometrial PGE(2) receptor (PTGER2) elevates the expression of enzymes involved in PGE(2) synthesis. Higher PGE(2) secretion in uterine lumen coincides with the elevated expression of HOXA10 transcription factor critical for implantation. A stable adhesion between conceptus and endometrium requires reduction in mucin-1 on the apical surface of epithelium and integrin activation by extracellular matrix proteins. Furthermore, growth factors, cytokines and its receptors are involved in embryo-maternal interactions.
Assuntos
Genitália Feminina/fisiologia , Prenhez , Suínos/fisiologia , Animais , Feminino , Inseminação Artificial/veterinária , Masculino , Gravidez , Prenhez/fisiologia , Prostaglandinas/metabolismo , SêmenRESUMO
Prostaglandins (PGs) play a pivotal role in maternal recognition of pregnancy and implantation in pigs. In the present study, PGE(2), PGF(2alpha), and PGFM (PGF(2alpha) metabolite) content, as well as PGE(2) synthase (mPGES-1) and PGF(2alpha) synthase (PGFS) expression was investigated in early pregnant gilts with natural (n=21) and PMSG/hCG-stimulated (n=19) estrus. Endometrial tissue samples, uterine luminal flushings (ULFs), and blood serum were collected on days 10-11, 12, and 15 after insemination. Additionally, day 15 conceptuses were collected for mPGES-1 and PGFS protein expression. Effect of estrus induction was observed on day 15 of pregnancy, when the content of PGE(2) in the uterine lumen was fourfold lower in gonadotropin-stimulated gilts in comparison to controls (P<0.001). Decreased PGE(2) content in ULFs of gonadotropin-treated pigs was preceded by lower endometrial mPGES-1 gene expression in hormonally-stimulated animals in comparison to control gilts (P<0.01). On the other hand, estrus induction with PMSG/hCG resulted in higher PGE(2) accumulation in the endometrial tissue on day 15 of pregnancy (P<0.01). Furthermore, PGF(2alpha) content in the endometrium and PGFM levels in blood serum were lower in gonadotropin-treated gilts, especially on day 12 after insemination when compared to control gilts (P<0.01). Finally, PGFS expression in day 15 conceptuses was decreased in animals with hormonally-induced estrus. We conclude that PMSG/hCG stimulation of prepubertal gilts to induce estrus results in changes of PG production and secretion during early pregnancy, which, in turn, may affect conceptus development, implantation, and the course of pregnancy.
Assuntos
Hidroxiprostaglandina Desidrogenases/genética , Oxirredutases Intramoleculares/genética , Indução da Ovulação/veterinária , Prostaglandinas/análise , Suínos/metabolismo , Útero/química , Animais , Gonadotropina Coriônica/administração & dosagem , Dinoprosta/análogos & derivados , Dinoprosta/análise , Dinoprosta/sangue , Dinoprostona/análise , Endométrio/anatomia & histologia , Endométrio/química , Endométrio/enzimologia , Estro , Sincronização do Estro , Feminino , Expressão Gênica , Gonadotropinas Equinas/administração & dosagem , Técnicas Imunoenzimáticas , Tamanho do Órgão , Indução da Ovulação/efeitos adversos , Indução da Ovulação/métodos , Reação em Cadeia da Polimerase , Gravidez , Prostaglandina-E Sintases , Prostaglandinas/biossíntese , RNA Mensageiro/análise , Útero/anatomia & histologia , Útero/enzimologiaRESUMO
Extended exposure of progesterone and conceptus estrogen influences the vascular compartment of the uterus and expression of many factors, such as prostaglandins (PGs), growth factors, extracellular matrix and adhesion molecules, cytokines and transcription factors. One of the supportive mechanisms by which the conceptus inhibits luteolysis is by changing PG synthesis in favor of luteoprotective PGE2. Alteration in PG synthesis may result from increased PGE synthase (mPGES-1) expression in the trophoblast and endometrium on days 10-13 of pregnancy with simultaneous down-regulation of PGF synthase (PGFS) and prostaglandin 9-ketoreductase (CBR1). Conceptus and endometrial, rather than luteal, synthesis of PGE2, is involved in the process of maternal recognition of pregnancy. However, complex (direct and indirect) actions of estrogen on the CL, including decreased luteal VEGF soluble receptor on day 12 of pregnancy, are important for luteal maintenance. Moreover, conceptus signals affect another lipid signaling component - lysophosphatidic acid receptor (LPA3), as well as HoxA10 and Wnt in the endometrium, to create the appropriate uterine environment for establishment of pregnancy and implantation.
Assuntos
Fase Luteal/fisiologia , Luteólise/fisiologia , Prenhez/fisiologia , Suínos/fisiologia , Animais , Implantação do Embrião/fisiologia , Endométrio/fisiologia , Estrogênios/fisiologia , Feminino , Gravidez , Progesterona/fisiologia , Prostaglandinas/fisiologia , Útero/fisiologiaRESUMO
Establishment of pregnancy in pigs requires estrogen secretion by the conceptus. The developmental changes of embryo before implantation and embryo-uterine cross talk are dependent on various biological molecules secreted by the endometrium and conceptus. An integral part of maternal recognition of pregnancy seems to be also the lipid signaling system consisting of prostaglandin (PG) F2 alpha and E2 and/or lysophosphatic acid (LPA). The downstream enzymes in PG synthesis pathway are: microsomal PGE synthase-1 (mPGES-1), PGF synthase (PGFS) and prostaglandin 9-ketoreductase/carbonyl reductase (CBR1) which catalyzes conversion of PGE 2 into PGF2 alpha. In contrast to mPGES-1, endometrial PGFS is highly increased on days 13-15 similarly as CBR1 on days 16-17 of the estrous cycle. Potential mechanism by which a conceptus inhibits luteolysis is changing the PGE2/PGF2 alpha ratio in favor of PGE2. It may be result of high expression of mPGES-1 in trophoblast and endometrium on days 10-13 of pregnancy and simultaneously the down-regulation of PGFS and CBR1 in conceptuses during this period. The conceptus can alter expression of endometrial CBR1 to modulate the PGE2/PGF2 alpha ratio in the uterus during the maternal recognition of pregnancy. High expression of conceptus and endometrial terminal PG synthases and CBR1 after initiation of blastocyst attachment suggest their involvement in early placentation. The higher LPA3 receptor mRNA expression during the early pregnancy compared to corresponding period of estrous cycle could indicate an important role of LPA and its receptor during the peri-implantation stage of pregnancy in pigs. Above results suggest that the lipid signaling system is an integral part of establishment and maintenance of pregnancy in the pig.
Assuntos
Lipídeos/fisiologia , Prenhez/fisiologia , Animais , Feminino , Fertilização/fisiologia , Hormônios/metabolismo , Gravidez , Prostaglandinas/fisiologia , SuínosRESUMO
Prostaglandins (PGs) of luteal origin may have paracrine and/or autocrine actions on the functions of the corpus luteum (CL). Previously, we have shown that enzymes of PG synthesis pathway such as prostaglandin E synthase (mPGES-1), prostaglandin F synthase (PGFS) and prostaglandin 9-ketoreductase (CBR1) are important in regulation of PG production in the conceptuses and endometrium of cyclic and pregnant pigs. Therefore, localization and expression patterns of these enzymes were determinated in porcine CL. The PGFS protein content was lower in metestrus and higher around luteolysis, and then decreased in late regressing CL. PGFS protein levels were lower on days 5-8 of pregnancy and did not differ between days 10 and 25. Elevated expression of mPGES-1 mRNA was found in early luteal phase. The mPGES-1 protein content, similarly to PGFS, was higher during luteolysis. mPGES-1 mRNA and protein levels were constant between days 5 and 25 of pregnancy. PGFS and mPGES-1 expression was down-regulated on days 16-17 of the oestrous cycle when compared to the corresponding days of pregnancy. Enhanced mPGES-1/PGFS ratio occurred during early luteal phase and days 5-8 of pregnancy. Expression of CBR1 mRNA and protein was constant during the cycle and pregnancy. Our studies revealed higher mPGES-1/PGFS ratios in the CL during early luteal phase and corresponding days of pregnancy that could favor PGE(2) synthesis and may be important in the control of luteal development. However, PG synthesis in the endometrium/conceptus rather than in the CL could be involved in luteolysis and maternal recognition of pregnancy in pigs.
Assuntos
Corpo Lúteo/metabolismo , Ciclo Estral/fisiologia , Regulação da Expressão Gênica/fisiologia , Prostaglandinas/biossíntese , Animais , Feminino , Perfilação da Expressão Gênica/veterinária , Gravidez , Prenhez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Suínos , Fatores de TempoRESUMO
We determined the expression of PGE2 synthase (mPGES-1), PGF synthase (PGFS), carbonyl reductase/prostaglandin 9-ketoreductase (CBR1) genes and the content of PGE2, PGF2alpha in porcine corpora lutea on Days 12-14 of pregnancy and Days 12-14 of the estrous cycle. For this study we used a surgically-generated model in which one of the uterine horns was cut transversely and a part of this horn was detached from the uterine corpus. The expression of mPGES-1, PGFS, and CBR1 genes and mPGES-1/PGFS ratio were significantly higher in corpora lutea of the pregnant gilts compared to the corpora lutea from the parallel ovaries of the cyclic gilts. There was no difference in mPGES-1, PGFS, CBR1 genes expression and mPGES-1/PGFS ratio between corpora lutea ipsi-(CL1) and contralateral (CL2) to the uterine horn with the developing embryos. The highest content of PGE2 was found in CL1 of the pregnant gilts. The PGE2/PGF2alpha ratio was significantly higher in CL1 of the pregnant gilts compared to corpora lutea from parallel ovary of the cyclic gilts. We suggest that the activity of the investigated genes is induced by compounds of embryonic origin which are not distributed only to the ipsilateral ovary but are transported within the mesometrium to both ovaries in a more systemic manner.
Assuntos
Blástula/fisiologia , Corpo Lúteo/fisiologia , Dinoprosta/metabolismo , Dinoprostona/metabolismo , Implantação do Embrião/fisiologia , Embrião de Mamíferos/fisiologia , Prostaglandinas/biossíntese , Animais , Primers do DNA , Estradiol/metabolismo , Feminino , Gravidez , RNA/genética , RNA/isolamento & purificação , DNA Polimerase Dirigida por RNA , Suínos , Útero/fisiologiaRESUMO
Improvement of cancer treatment is a major challenge of medical research. Despite the immense efforts made in the improvement of diagnosis and treatment, cancer remains a major concern and cause of morbidity and mortality. Most of the modern anti-neoplastic therapies have severe side effects, and tumor cells often develop drug resistance. There is promise in the new generation of treatments (gene therapy, immunotherapy, vaccines, etc.) that are under development, but the efficacies and side effects of such therapies have so far been disappointing. Receptor-based therapies are not new, but many normal cells also present the same receptors reducing the specificity of such approaches. Several lytic peptides have been investigated because of they appear to kill cancer cells due to changes of their membrane potential. Thus, linking receptor-specific ligands to lytic peptides is expected to augment the specificity of targeting and decrease the toxicity of lytic peptides on normal cells. One such polypeptide is hecate (an analogue to the bee venom main component, melittin) that preferentially kills cancer cells at low doses. When this peptide is fused with the 81-95 amino acid fragment of chorionic gonadotropin-beta (CGbeta) subunit (hecate-CGbeta), it targets cells expressing luteinizing hormone receptor (LHR), even at very low doses, or when LHR is expressed at low level. Our recent data showed that this peptide conjugate is efficient in destroying LHR-positive cells in xenografts and more importantly in transgenic mouse models developing LHR-positive somatic cell tumors in gonads. The mechanism of action of hecate-CGbeta after binding to LHR is destruction of cell membranes resulting in rapid cell death by necrosis with minimal side effects. This review summarizes our findings on the action of this novel peptide and considers the future potential of this family of targeting peptides in the treatment of neoplasias.
Assuntos
Meliteno/análogos & derivados , Neoplasias Ovarianas/tratamento farmacológico , Receptores do LH/metabolismo , Neoplasias Testiculares/tratamento farmacológico , Animais , Gonadotropina Coriônica Humana Subunidade beta/uso terapêutico , Sistemas de Liberação de Medicamentos , Feminino , Terapia Genética , Humanos , Masculino , Meliteno/uso terapêutico , Modelos Biológicos , Neoplasias Ovarianas/metabolismo , Receptores do LH/antagonistas & inibidores , Receptores do LH/genética , Neoplasias Testiculares/metabolismoRESUMO
Inhibition of luteolysis and establishment of pregnancy in pigs results from oestrogen secretion by the conceptuses and requires progesterone produced by the corpus luteum (CL). An integral part of maternal recognition of pregnancy in the pig is the redirection of prostaglandin (PG) F2alpha secretion from endocrine (blood) to exocrine (uterus) direction and an increase of PGE2 synthesis in both the endometrium and conceptus. Uterine and conceptus PGE2 synthases play an integrated role in establishing the PGE2:PGF2alpha ratio necessary for luteal maintenance. The luteolytic or luteotrophic changes in the CL are synchronised with the release of maternal pituitary and ovarian hormones. The presence of uterine oxytocin (OT) and luteinising hormone (LH) receptors are important for the luteolytic effect of PGF2alpha. Conceptus oestrogen secretion coincides with autocrine and paracrine dialogue between the multiple conceptuses and uterine biological compounds and their receptors in trophoblast and endometrium.
Assuntos
Implantação do Embrião/fisiologia , Embrião de Mamíferos/metabolismo , Manutenção da Gravidez , Prenhez/metabolismo , Suínos/metabolismo , Útero/metabolismo , Animais , Feminino , Hormônios Esteroides Gonadais/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Gravidez , Prostaglandinas/metabolismoRESUMO
Cyclooxygenase (COX) is the rate-limiting enzyme that catalyses the initial step in prostaglandins (PGs) production. In the present studies, endometrial COX-1 and COX-2 expression throughout the oestrous cycle and early pregnancy was analysed in pigs using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry. There were no changes in messenger RNA (mRNA) and protein expression for COX-1 in cyclic pigs. In pregnant animals, mRNA levels of this enzyme increased on days 22-25 (p < 0.001). However, no upregulation of COX-1 protein was detected. Quantification of COX-2 mRNA expression during the oestrous cycle revealed significant increases on days 10-12 and 14 (p < 0.001 and p < 0.01 vs days 2-4, respectively). Protein levels were also increased on day 14 when compared with days 2-12 and 18-20 after oestrus. In pregnant animals, the patterns of both COX-2 mRNA and protein expression were similar. Messenger RNA levels were higher on days 16 and 22-25 (p < 0.01 vs day 10). Moreover, the protein content tended to increase on days 16 and 22-25. COX-1 and COX-2 were localized in the luminal and glandular epithelium as well as in the uterine stroma. In contrast to COX-1, a positive immunostaining reaction for COX-2 was detected only on days 12-16 after ovulation and on days 14-16 of pregnancy. In conclusion, these results indicate specific patterns of COX-1 and COX-2 expression in the porcine endometrium throughout the oestrous cycle and early pregnancy. COX-2 rather than COX-1 seems to be the primary enzyme responsible for modulated PGs production at the time of luteolysis in cyclic and during implantation in pregnant animals.
Assuntos
Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Endométrio/enzimologia , Ciclo Estral/metabolismo , Prenhez/metabolismo , Suínos , Animais , Western Blotting/veterinária , Ciclo-Oxigenase 1/genética , Ciclo-Oxigenase 2/genética , Feminino , Imuno-Histoquímica/veterinária , Gravidez , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterináriaRESUMO
Luteinising hormone (LH) and human chorionic gonadotropin (hCG) share a common receptor in gonadal cells. The receptors have also been detected in several nongonadal but reproduction-associated tissues of pigs, cattle, and other species including the uterus (myometrium, endometrium), oviduct, cervix, blood vessels, mammary gland and other tissues. The main role of LH/hCG receptors in the myometrium is stimulation of growth and hyperplasia, and relaxation of uterine motility; hCG also boosts blood flow in the uterine artery. LH and hCG can increase production of prostaglandins in the endometrium, oviduct, and blood vessels. We suggest that the preovulatory surge of LH plays an important role in controlling oviductal contractions. Awareness of LH binding to many tissues of the female reproductive tract and integration with embryonic factors may lead to the elaboration of new strategies for improved reproductive efficiency in domestic species. Mammary glands also possess LH/hCG receptors through which gonadotropins can affect the metabolism of steroid hormones and could play an inhibitory role in mammary carcinogenesis and in the growth of breast tumours. A novel approach to target and ablate carcinoma cells through LH receptors is described.
Assuntos
Tubas Uterinas/metabolismo , Hormônio Luteinizante/fisiologia , Glândulas Mamárias Animais/metabolismo , Receptores do LH/metabolismo , Reprodução/fisiologia , Útero/metabolismo , Animais , Mama/metabolismo , Colo do Útero/metabolismo , Gonadotropina Coriônica/metabolismo , Feminino , Hormônios Esteroides Gonadais/metabolismo , Humanos , Hormônio Luteinizante/metabolismo , Útero/irrigação sanguíneaRESUMO
Motor fatigue, during 30 seconds of maximum voluntary isometric contraction (MVIC) was simultaneously evaluated by the decline in mechanical force output, and from the compression in the power spectrum obtained from surface electromyogram (sEMG). Measurements were performed in patients diagnosed with amyotrophic lateral sclerosis (ALS) and normal control (NC) in two muscle groups, elbow flexors (EF) and ankle dorsiflexors (DF). The decline in force output, as a manifestation of mechanical fatigue, was digitally calculated online by partitioning the force versus time curve to determine the percent of MVIC reduction over a 30 sec period and was expressed as force fatigue index (FFI). The compression in the sEMG power spectrum, as a manifestation of myoelectrical fatigue, was tracked by calculating the median frequency shift (MFS) from the first 5 sec to the last 5 sec of the 30 sec MVIC using digital Fast Fourier Transformation. In ALS patients, the significantly higher reduction in mechanical force output during the 30 sec MVIC (higher FFI) was accompanied with significantly less compression in the sEMG power spectrum (less MFS) as compared to NC (P < or =0.005) in the two muscle groups. This dissociation between the mechanical and myoelectrical manifestation of muscle fatigue in ALS indicates that a reduction in muscle fiber conduction velocity (MFCV) may be a contributing peripheral factor in the pathogenesis of muscle fatigue in ALS. Alterations in motor unit functionality, especially in type II fast motor unit muscle fibers, and structural damage in denervated muscle fibers may contribute to the lower MFCV during motor fatigue in ALS patients.
Assuntos
Esclerose Lateral Amiotrófica/fisiopatologia , Contração Isométrica/fisiologia , Fadiga Muscular/fisiologia , Adulto , Idoso , Tornozelo/fisiopatologia , Estudos de Casos e Controles , Cotovelo/fisiopatologia , Eletromiografia/métodos , Teste de Esforço , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fibras Musculares Esqueléticas/fisiologia , Esforço FísicoRESUMO
Motor fatigue is a common complaint in patients with amyotrophic lateral sclerosis (ALS), but is often excluded, unlike weakness, from the clinical assessment of these patients. This could be due to the complexity and often painful assessment techniques of this motor deficit. This study examines the feasibility of quantitative assessment of motor fatigue by modifying presently available force measurements. The relationship between weakness and fatigue in ALS patients was also examined. Fifty-four ALS patients and 39 normal control subjects performed 30 s of sustained maximal voluntary isometric contraction (MVIC) of elbow flexors (EF), knee extensors (NE), and ankle dorsiflexors (DF), using a computerized force measurement system and standardized testing procedures. Fatigue index (FI) was digitally calculated, from the force-time curve, as the percentage of MVIC unable to be sustained over the 30-s period. Fatigue was greater in ALS patients than in normal control (mean=23% vs. 15%) in all muscles including muscles that were not clearly weak. Weakness and fatigue were poorly correlated in ALS patients and may be independent measures of the pathogeneses of ALS.
Assuntos
Esclerose Lateral Amiotrófica/fisiopatologia , Fadiga Muscular , Esclerose Lateral Amiotrófica/diagnóstico , Tornozelo/fisiopatologia , Área Sob a Curva , Cotovelo/fisiopatologia , Teste de Esforço , Estudos de Viabilidade , Humanos , Contração Isométrica , Joelho/fisiopatologia , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Índice de Gravidade de DoençaRESUMO
Porcine pregnancy-associated glycoprotein genes (pPAG) are known as a multigene family, in which five members have been cloned and sequences of their cDNAs identified. Porcine PAG1 and pPAG3 genes, belonging to the pPAG1-like subfamily, both encode enzymatically inactive precursors. In contrast, cDNAs of pPAG2, pPAG4 and pPAG6 represent the pPAG2-like gene subfamily, encoding enzymatically active precursors. The objective of this study was to investigate the polymorphism of both pPAG-like gene subfamilies in the pig in comparison to other domestic species, including cattle, sheep and goat (Artiodactyla), their wild relatives (red deer and wild pig) and horse (Perissodactyla). This is the first paper indicating the polymorphism of the pPAG gene family, examined by lengths of amplified genomic fragments (PCR). Obtained PCR products were analysed in relation to five characterised cDNAs of pPAGs (pPAG1-like and/or pPAG2-like subfamilies) and according to one recognised structural exon-intron organisation of the pPAG2 gene, among at least eight pPAG2-like genes expected in the porcine genome. The highest polymorphism frequency of both pPAG1- and pPAG2-like gene subfamilies was found in the second region, exons 5 and 6 (with intron E). The length of PCR-amplified genomic fragments was approximately: 1043, 700, 600 and 193 bp. A high polymorphism frequency was found in the 3'-terminal fragment, corresponding to exons 7-9 (with introns G and H), more frequent the pPAG2-like gene subfamily. The length of PCR-amplified genomic fragments was approximately: 733, 650 and 356 bp. In contrast, PAG polymorphism was not detected in another region, encompassing exons 2-4 (with introns B and C). The length of PCR-amplified genomic fragments was approximately 279 bp in all examined genomes. In conclusion, amplification of various regions of the PAG gene family presents a relatively inexpensive PCR method of animal pre-selection with different genotypes. Such a pre-selection of animals is helpful for further gene number inquiry of the PAG gene family in each animal, then in related generations. The obtained results provide a useful background for a genetic marker preparation (by Southern analysis of the PAG family) that will presumably enable an economical early selection of young animals for effective reproduction.
RESUMO
Botulism in infants, unless recognized early, is associated with high mortality and morbidity. The diagnosis is suspected when infants present with sudden onset of weakness, respiratory failure, and constipation and is confirmed by demonstration of botulinum toxin in stool several weeks later. Electrodiagnosis allows quick and reliable confirmation of botulism. Low-amplitude compound muscle action potentials, tetanic or post-tetanic facilitation, and the absence of post-tetanic exhaustion support the diagnosis. Two infants with confirmed botulism did not exhibit the characteristic electrodiagnostic features, demonstrating the pitfalls in electrodiagnosis of infantile botulism.
Assuntos
Botulismo/diagnóstico , Erros de Diagnóstico , Eletrodiagnóstico/métodos , Biópsia , Clostridium botulinum/isolamento & purificação , Fezes/microbiologia , Feminino , Humanos , Lactente , Músculos/patologiaRESUMO
A renal transplant patient developed chronic and progressive back and lower extremity pain followed by foot weakness. The correct diagnosis of lumbosacral plexopathy was made after electromyography and nerve conduction studies and the etiology of radiculopathy due to nerve root compression was excluded. This prompted further investigations that led to the discovery of a large internal iliac artery pseudoaneurysm. We emphasize the use of electrodiagnostic studies to investigate patients with back and limb pain for correctly localizing responsible pathology. In this case a potentially lethal situation was correctly identified in a transplant patient.
Assuntos
Falso Aneurisma/etiologia , Artéria Ilíaca , Transplante de Rim , Plexo Lombossacral , Doenças do Sistema Nervoso/etiologia , Complicações Pós-Operatórias , Falso Aneurisma/complicações , Falso Aneurisma/diagnóstico , Dor nas Costas/diagnóstico , Dor nas Costas/etiologia , Eletrodiagnóstico , Humanos , Perna (Membro) , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Doenças do Sistema Nervoso/diagnóstico , Dor/diagnóstico , Dor/etiologiaRESUMO
We compared quantitatively the myotoxicity of 3'-azido-2',3'-dideoxythymidine (AZT) against uninfected and ts1 retrovirus infected mouse skeletal muscle (ATCC CRL 1772) cells at different stages of maturation in vitro. The AZT half inhibitory concentration (IC50) for myoblast proliferation was determined for uninfected myoblasts and parallel cultures infected with ts1 virus. The AZT IC50 for muscle cell differentiation was determined in cultures where myoblasts were grown to confluence and then changed to the fusion medium to which AZT was added at increasing concentrations. Creatine kinase activity was used as a marker of muscle cell differentiation and was determined in homogenates after 7 days. Total cellular mitochondrial DNA was analyzed by Southern blotting. The estimated AZT IC50 for muscle cell proliferation (2-5 microM) was significantly less than the AZT IC50 for muscle cell differentiation (100 microM). Infection with ts1 retrovirus did not significantly shift the IC50 for either proliferation or differentiation of muscle cells. Toxic concentrations of AZT did not cause selective depletion of mitochondrial DNA. The myotoxic effects of AZT on myoblast proliferation and muscle cell differentiation in vitro were quantitatively different and were not changed by productive ts1 retrovirus infection of muscle cells. These results suggest that AZT may impair muscle fiber regeneration in the course of retrovirus associated myopathy. The mechanism of AZT myotoxicity was not explained by alterations in total mitochondrial DNA content.