RESUMO
This systematic review aimed to update the perceived needs of individuals with breast cancer (BC). Databases were searched for studies reporting quantitative data collected through validated assessment tools. Needs of adults with BC were reported by survivorship phase. The post-diagnosis and the post-surgery phases revealed the most needs; health system and information needs represented the greatest concern, with average Supportive Care Needs Survey-Short Form (SCNS-SF34) scores ranging from 62.0 to 75.8 post-diagnosis and from 45.0 to 67.8 post-surgery. Needs then seemed to decrease or remain stable up to within one year from diagnosis, when needs in all domains increased again; health system and information needs remained a priority. Younger age, side effects, type of treatment, and advanced stage were associated with the occurrence of unmet needs. The needs of BC survivors vary over the course of their cancer experience. This knowledge can assist the planning of appropriate assessments.
Assuntos
Neoplasias da Mama , Sobreviventes de Câncer , Feminino , Humanos , Neoplasias da Mama/terapia , Neoplasias da Mama/patologia , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/epidemiologia , Sobreviventes de Câncer/estatística & dados numéricos , Necessidades e Demandas de Serviços de Saúde , Avaliação das NecessidadesRESUMO
In neonatology, blood gas analysis is a useful tool in the evaluation of the health of newborns and plays a key role in early detection of critically ill subjects. Because blood gas analysis parameters have not previously been studied in any depth in donkey foals, this study was performed on 16 healthy Martina Franca donkey foals born after an uncomplicated delivery. Arterial and venous blood samples were collected at 5 minutes and at 12, 24, 72, and 96 hours of age. Blood gas analysis was performed by a portable analyzer, measuring arterial and venous total carbon dioxide, carbon dioxide partial pressure (pCO2), oxygen partial pressure (pO2), oxygen saturation (sO2), bicarbonate, base excess (BE), pH, and lactate (LT). Lower blood pH values, pO2 and sO2, and a higher level of lactate were found at birth in comparison with subsequent sampling times. This moderate acidotic profile disappeared at 12 hours, when all the parameters became constant until the end of the study period. As expected, significant differences between arterial and venous blood gas parameters related to the oxygenation, such as pO2 and sO2, and partially carbon dioxide partial pressure were found, whereas total carbon dioxide, pH, BE, and LT were comparable in arterial and venous blood samples. For these latter parameters, the highly significant correlation between arterial and venous findings suggests that venous samples could be an acceptable alternative to the arterial sample for blood gas analysis in newborn donkey foals, when the oxygenation status of the patient is not the first goal of patient analysis.
Assuntos
Animais Recém-Nascidos/sangue , Dióxido de Carbono/sangue , Equidae/sangue , Oxigênio/sangue , Animais , Animais Recém-Nascidos/fisiologia , Bicarbonatos/sangue , Equidae/fisiologia , Concentração de Íons de Hidrogênio , Lactatos/sangueRESUMO
In this study, the first postpartum heat, termed the foal heat, characteristics and performance in female donkey (jenny) of Martina Franca are described. To this end, the follicular development of 42 jennies during foal heat was compared with that of 31 jennies at the third estrus after foaling. Estrus length (7.1±0.9 and 6.8±0.7 days), follicular development and preovulatory follicle size (43.7±3.5 and 45.1±2.5 mm) were similar between jennies during the foal heat and during the third estrus after foaling. The pregnancy rate at day 14 was significantly lower in the foal heat jennies (57.1%) than the third estrus jennies (82.3%). However, the pregnancy rate at day 14 in foal heat jennies increased significantly when the onset of foal heat was ⩾8 days after foaling (93.8%) or when the ovulation happened ⩾12 days after foaling (85.7%). The data provided in the present study suggest that the foal heat in the endangered jennies of Martina Franca could be successfully utilized to reduce the interpartum period if the onset of foal heat occurs >8 days after foaling.
Assuntos
Equidae/fisiologia , Estro/fisiologia , Período Pós-Parto/fisiologia , Complicações na Gravidez/veterinária , Reprodução , Animais , Temperatura Corporal , Feminino , Folículo Ovariano , Ovulação , Gravidez , Taxa de Gravidez , Distribuição Aleatória , Fatores de TempoRESUMO
In the recent years, the donkey population decreased dramatically so that many breeds are presently considered as endangered. In comparison to the horse, the donkey placenta still remains not completely studied. In the horse, one of the diagnostic tools useful to identify pregnant mares at risk of abortion or premature delivery, include the transrectal ultrasound examination of the uterus and its contents; and especially of the combined thickness of the uterus and of the placenta (CUPT). Since the CUPT was never investigated in donkeys, the present study was aimed to define the transrectal CUPT values during the last half of pregnancy in 20 Martina Franca jennies. Foalings times, foals characteristics and placental gross appearance, and measurements were also evaluated and values resulted always within normality. Differently to the mare, a continuous significant CUPT increase between the sixth to the 12 months of pregnancy, and a substantial increase from the ninth to the 12th month of pregnancy, was found. Although statistically not evaluable, the CUPT values recorded from three jennies with pregnancy loss did not show evidence of CUPT increases. In conclusion, normal CUPT values from the sixth to the 12th month of pregnancy in Martina Franca donkeys are provided, but further investigations are needed to define possible breed or body-size CUPT specific differences, as well as the CUPT values during pregnancy disturbances or placental abnormalities.
Assuntos
Equidae/fisiologia , Placenta/diagnóstico por imagem , Prenhez , Ultrassonografia/veterinária , Útero/diagnóstico por imagem , Animais , Feminino , Gravidez , Prenhez/fisiologiaRESUMO
Equine spermatozoa from the cauda epididymis were previously collected and frozen, and the fertility was assessed. Most studies were performed on healthy stallions that had undergone routine castration or on the epididymis collected at the abattoir, but there are no studies on the quality of epididymal semen in subjects which have died from colic or which underwent intensive care. The present study was designed to verify whether a severe illness could affect epididymal semen quality and freezability in the stallion. Therefore, epididymal semen characteristics during the freezing process in stallions which had died from colic and in healthy stallions submitted to elective castration were compared. Five stallions that had died from colic (ill stallions [ISs]) and seven stallions that had undergone elective castration (healthy stallions) were castrated, and cauda epididymis spermatozoa were collected and processed. Sperm quality was tested after collection, after washing procedures, at the end of the equilibration (5 °C for 75 minutes), and after freezing/thawing. Sperm quality was measured by objective motility characteristics, membrane and acrosome integrity, and mitochondrial activity. After collection, sperm in ISs showed low kinetic parameters (total motility: 17.3 ± 3%, progressive motility: 6 ± 1%, average path velocity: 57.4 ± 35.4 µm/s, straightness: 74.2%) compared with healthy stallions (total motility: 90.8 ± 3.7%, progressive motility: 70 ± 4%, average path velocity: 118.1 ± 12.6 µm/s, straightness: 82.4%) but demonstrated similar membrane and acrosome integrity (85 ± 2.8% vs. 87.6 ± 3.1%). Sperm kinetic parameters increased after washing procedures and cooling in ISs, reaching comparable values after equilibration (5 °C for 75 minutes) and freezing/thawing. The data reported in this study suggest that the quality of the equine epididymal spermatozoa cryopreserved in stallions that had died from colic was similar to that reported in epididymal sperm after elective castration and was also similar to the data reported in literature for cryopreserved equine semen.
Assuntos
Cólica/veterinária , Criopreservação/veterinária , Epididimo/citologia , Doenças dos Cavalos/fisiopatologia , Preservação do Sêmen/veterinária , Espermatozoides/fisiologia , Acrossomo/fisiologia , Animais , Membrana Celular/fisiologia , Cólica/mortalidade , Cólica/fisiopatologia , Cavalos , Masculino , Mitocôndrias/fisiologia , Orquiectomia/veterinária , Análise do Sêmen/métodos , Análise do Sêmen/veterinária , Preservação do Sêmen/métodos , Motilidade dos Espermatozoides , Espermatozoides/ultraestruturaRESUMO
The knowledge of reference ranges for hematologic, biochemical, and blood gas parameters in the different species and the influence of breed and age on them is a fundamental tool for the clinician. For this reason, the aim of this study was to evaluate the age-related changes of hematologic and biochemical parameters in Martina Franca donkey foals during the first 3 weeks of life and of blood gases during the first 24 hours of age. Fifteen healthy donkey foals were enrolled; blood samples were collected from each foal at 10 minutes after birth, 1 hour after the first and second suckles, 12 and 24 hours after birth, daily from Day 2 to 7, and at Days 10, 14, and 21 of life. Erythrocytes, leukocytes, and platelets counts were assessed; also metabolic (alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, creatinphospokinase, lactate dehydrogenase, alkaline phosphatase, glucose, blood urea nitrogen, creatinine, total proteins, albumins, cholesterol, and total bilirubin) and electrolytic parameters (Ca, P, Mg, Na, K, and Cl) were evaluated. Finally, blood gases and metabolic parameters (pH, pCO2, pO2, sO2, TCO2, HCO3, lactate, and base excess) on venous blood were assessed with a portable analyzer. A statistical analysis to evaluate the influence of age and sex was performed. Several differences were found between sampling times, demonstrating that age influences these parameters. Moreover differences were found compared with data reported in literature for donkey foals of another species, horse foals, and adult donkeys. Although a great interindividual variation for some parameters exists, this study demonstrated that interval references should be addressed not only to different species, but also to specific breeds and to the neonatal period.
Assuntos
Animais Recém-Nascidos/sangue , Equidae/sangue , Animais , Contagem de Células Sanguíneas , Análise Química do Sangue/veterinária , Gasometria/veterinária , Feminino , MasculinoRESUMO
The Martina Franca (MF) donkey breed, with 48 jackasses and 515 jennies, is considered an endangered breed according to the data from the Monitoring Institute for Rare Breeds and Seeds in Europe. The knowledge of the estrous cycle characteristics has a great impact for assisted reproduction, especially in endangered species. In this study, the estrous cycle characteristics were investigated in 12 MF jennies throughout the year. Estrous cycle, estrous and diestrous lengths, follicular growth and ovulation, and estradiol-17ß (E2) and progesterone (P4) plasma concentrations were monitored in MF jennies and compared in different seasons. In all jennies (100%) estrous cycle was detected during the whole year, with no differences in the estrous cycle length among seasons. However, a significant increase of estrous length in spring and summer compared with autumn and winter was found. Diestrus was shorter in summer than in the other seasons. Estrous behavior was always shown and characterized by rhythmic eversion of the vulvar labia (winking) with exhibition of the clitoris, urination, male receptivity and clapping, with sialorrhoea, neck and head extension, and back ears. Estrus was characterized by the ovulation of a larger follicle in spring and summer than in autumn and winter. The pattern of E2 and P4 plasma concentrations during the estrous cycle were similar to that reported for the mare, but without differences among the four seasons, so that a negligible effect of environmental conditions on ovarian E2 and P4 secretion was hypothesized, despite the larger diameter of the ovulating follicle in spring and summer.
Assuntos
Equidae/fisiologia , Ciclo Estral/fisiologia , Folículo Ovariano/crescimento & desenvolvimento , Estações do Ano , Animais , Diestro/fisiologia , Espécies em Perigo de Extinção , Estradiol/sangue , Estro/fisiologia , Feminino , Masculino , Ovulação/fisiologia , Progesterona/sangue , Reprodução , Técnicas de Reprodução Assistida/veterinária , Comportamento Sexual Animal , Fatores de TempoRESUMO
In this study, the effect of the chamber used for the automated analysis of sperm motility by a computer-assisted semen analysis system on sperm kinematics was evaluated, and the cause of this effect was also verified. Twenty-three bull semen batches were thawed, and semen was diluted, aliquoted and analysed with six different chambers, (three capillary-loaded chambers and three droplet (DR)-loaded chambers). For each chamber type, each sample was analysed in quadruplicate, and the reliability of the analysis was tested using an intraclass correlation coefficient (ICC). Furthermore, sperm membrane integrity (MI) was evaluated, for each sample and chamber, in 12 randomly selected central and 12 edge fields. The ICC analysis showed that some parameters could have a significant variability related to the chamber. High stability of results was detected in Leja 4-chamber slide. Furthermore, as previously reported in other studies, capillary-loaded chambers seemed to affect the total and progressive motility and sperm velocities (average path velocity, straight line velocity and curvilinear velocity). These findings were corroborated by the evaluation of sperm MI that was significantly higher in the DR-loaded chambers. This study confirms that the chamber used for the objective kinetic evaluation of bull-thawed spermatozoa significantly affects the result. These differences could be present also in other species, even if the specific effect on the sperm kinematics should be verified. The Makler chamber seemed to give reliable results with negligible effects on sperm kinematics.
Assuntos
Criopreservação , Análise do Sêmen/instrumentação , Motilidade dos Espermatozoides , Animais , Bovinos , Masculino , Análise Numérica Assistida por Computador , Análise do Sêmen/métodos , Preservação do Sêmen/métodos , EspermatozoidesRESUMO
As other European donkey breeds, Martina Franca could be considered an endangered breed because of the population number (48 jackasses and 515 jennies in 2011). To increase donkey population and breed biodiversity preservation, several research projects on donkey reproduction have been performed; however reproductive seasonality has been only partially investigated in jackasses. For this reason the aim of this study was to investigate the effect of seasons and of long (spring to summer; SS) and short (autumn to winter; AW) day length periods, on reproductive physiology of Martina Franca jackasses, in particular on: (1) testicular morphometric characteristics, (2) behavior, through the evaluation of the reaction time, and finally, (3) semen characteristics. Seven adult and reproductively mature Martina Franca jackasses were enrolled. For each jackass, a morphometric evaluation of both testes was performed once for every season, before the first seasonal collection. Semen was collected, by artificial vagina, weekly for a whole year (52 collections per donkey); at each collection the reaction time was recorded and a complete semen evaluation was performed immediately after collection. No differences in testicular measures were observed neither between left and right testis nor during the four seasons. A lower reaction time was observed in spring and summer compared with autumn and winter and during the period SS compared with AW. Total volume was significantly higher in winter compared with all the other seasons; gel-free volume was higher in winter compared with summer and autumn. Mean sperm concentration was significantly lower in winter compared with spring and summer and in AW compared with SS. Total and progressive motility and membrane integrity did not show any significant difference between season and between SS and AW. A lower average path velocity was observed in autumn compared with spring and summer, and in summer a higher straight line velocity compared with spring and autumn, and a lower curvilinear velocity compared with winter were found. No differences in amplitude of lateral head displacement, beat cross frequency, straightness and linearity were observed neither between seasons nor in SS versus AW. Seminal pH did not show any seasonal difference; also sperm morphology did not show any significant difference during the whole year. This study demonstrated that under our climatic and management conditions Martina Franca donkey stallions do not show significant differences in testicular morphometric characteristics during the year, even if a certain seasonal influence exists on reaction time and some seminal parameters. Considering the variable temperature and photoperiod during the four seasons where the study was performed, this lack of substantial differences could be related to a characteristic of the Martina Franca breed.
Assuntos
Equidae/anatomia & histologia , Equidae/fisiologia , Estações do Ano , Sêmen/fisiologia , Testículo/anatomia & histologia , Animais , Masculino , Ereção Peniana/fisiologia , Fotoperíodo , Tempo de Reação , Sêmen/química , Contagem de Espermatozoides , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/anormalidades , TemperaturaRESUMO
In mammals, the epididymis has numerous interrelated functions including absorptive and secretory activity that affect luminal environment and cell membrane, and the maturation and storage of sperm. Spermatozoa acquire their motility and fertilizing ability during their passage through the epididymis and the motility of epididymal spermatozoa should be a balance between the maturation of flagellum and the inhibition of the flagellar machinery. In this study maturational change in sperm characteristics were evaluated in the epididymis of donkey. Spermatozoa collected from four portions of the epididymis (head, cranial corpus, caudal corpus, tail) were compared before and after ejaculation for viability, mitochondrial activity, kinetic parameters, and morphology. A significant increase in the mitochondrial activity along the epididymis was reported, suggesting a possible involvement in the motion mechanism. This should be corroborated by the significant correlation between mitochondrial activity and the total and progressive motility and the increase in velocities of spermatozoa recorded by computer-assisted sperm analysis. The percentage of most of the abnormal spermatozoa were similar in all tracts, with a great variability between jackasses. Only the bent midpiece percentage decreased significantly along epididymis. A significant increase in the percentage of distal cytoplasmic droplets (DCD), and a simultaneous decrease in the proximal cytoplasmic droplets (PCD), was found. The DCD fell down after ejaculation suggesting the late loss of the cytoplasmic residual (DCD) in the donkey, as hypothesized in the stallion. Because the prevalence of PCD were similar in both tail epididymal and ejaculated spermatozoa, a defect of the maturative process in the PCD sperm should be speculated.
Assuntos
Epididimo/citologia , Equidae , Espermatozoides/citologia , Animais , Masculino , Mitocôndrias/fisiologia , Análise do Sêmen/veterinária , Motilidade dos Espermatozoides , Espermatozoides/crescimento & desenvolvimentoRESUMO
The aim of the study was to compare horse and donkey placentae using stereological techniques. Term placentae were collected at spontaneous foaling from seven Thoroughbred mares, seven pony mares, and six jenny donkeys. Maternal and foal weights were recorded and the mass, volume, and gross area of each allantochorion was also recorded. Ten random biopsies were recovered and processed for light microscopy from which the surface density of the microcotyledons (S(v)) and the total microscopic area of fetomaternal contact were calculated stereologically. Gestation length was longer in the donkeys than the other two groups (median values: 371 vs. 327 and 341 days, P < 0.05). There were significant correlations between foal birthweight and gross area (rho = 0.89; n = 20; P < 0.05), mass (rho = 0.84; n = 20; P < 0.05) and volume (rho = 0.89; n = 20; P < 0.05) of the allantochorion. S(v) was higher in the donkey placenta than the other groups (median values: 0.05 vs. 0.03 and 0.04 microm(-1), P < 0.05) although placental efficiency was lower in the donkeys (median values: 0.87 vs. 1.33 and 1.32 kg/m2, P < 0.01). The results of the study confirmed that, although strong morphological similarities exist between the allantochorion of the horse and donkey, that of the donkey develops more complex microcotyledons, as judged stereologically, and exhibits a lower placental efficiency. These differences may be related to maternal genotype and/or the longer gestation length shown by the donkey compared to the horse, but a negative correlation (rho = -0.92, P < 0.01) was also found between age and placental efficiency in donkeys.
Assuntos
Equidae/anatomia & histologia , Placenta/anatomia & histologia , Prenhez/fisiologia , Animais , Tamanho Corporal , Feminino , Placenta/fisiologia , GravidezRESUMO
GLUTs are a family of proteins that facilitate the transport of glucose and other hexoses through the plasma membrane of the cells. GLUTs are present in mammalian spermatozoon's membrane in different isoforms and they supply metabolic substrates for all the cell's activities such as motility, homoeostasis and fertilization. As studies about donkey spermatozoa and their metabolism are lacking, this study was aimed at detecting GLUTs 1, 2, 3 and 5 presence by western blotting technique and at determining their localization on the plasma membrane by indirect immunofluorescence. Each protein showed a typical localization on the sperm cells' plasma membrane, differencing the one to the other on the basis of the hexose they transport. We also highlighted some differences between GLUTs distribution and molecular weight in donkey spermatozoa and its nearest relative, the horse.
Assuntos
Equidae/metabolismo , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Transporte Proteico/fisiologia , Espermatozoides/metabolismo , Animais , Membrana Celular/metabolismo , Proteínas Facilitadoras de Transporte de Glucose/genética , Imunoquímica , Masculino , Motilidade dos Espermatozoides , Espermatozoides/citologiaRESUMO
A polymerase chain reaction (PCR)-based assay was used for the specific detection of Habronema microstoma and Habronema muscae (Nematoda, Spirurida) in order to identify the intermediate hosts of both nematode species under field conditions. A total of 1087 netted and 165 laboratory-bred flies were tested. Flies were identified as Musca domestica Linnaeus 1758, Musca autumnalis De Geer 1776, Haematobia irritans (Linnaeus 1758), Haematobia titillans (De Geer 1907) and Stomoxys calcitrans (Linnaeus 1758) (Muscidae). Genomic DNA was extracted from pools of fly heads, thoraces and abdomens, and 703 samples were subjected to a duplex two-step semi-nested PCR assay to specifically detect diagnostic regions within the ribosomal ITS2 sequence of both H. microstoma and H. muscae. Stomoxys calcitrans specimens were positive for H. microstoma DNA and M. domestica specimens were positive for H. muscae DNA. In particular, PCR-positive samples derived from both farm-netted and laboratory-bred flies. The present study represents the first evidence of the vectorial competence of different fly species as intermediate hosts of Habronema stomachworms under field conditions. We discuss the roles of S. calcitrans and M. domestica in transmitting H. microstoma and H. muscae.
Assuntos
Dípteros/parasitologia , Spiruroidea/fisiologia , Animais , Feminino , Interações Hospedeiro-Parasita , Insetos Vetores/parasitologia , Larva/parasitologia , Masculino , Reação em Cadeia da Polimerase , Pupa/parasitologiaAssuntos
Ejaculação/fisiologia , Equidae/fisiologia , Glândulas Seminais/diagnóstico por imagem , Animais , Lateralidade Funcional , Concentração de Íons de Hidrogênio , Masculino , Sêmen/fisiologia , Glândulas Seminais/anatomia & histologia , Glândulas Seminais/fisiologia , Espermatozoides/citologia , Espermatozoides/fisiologia , UltrassonografiaRESUMO
Native PGF(2alpha) and its analogs have been used in the horse mare to manipulate ovarian activity, primarily as luteolytic agents to induce estrus. Despite numerous studies on the effects of these luteolysins in the mare, to date only a single investigation has been conducted in the jenny. The aim of this study was to evaluate the response of the corpus luteum (CL) to a single dose of PGF(2alpha) given 3 days (72h) after ovulation and to establish the plasma progesterone (P4) profile from pre-treatment to post-treatment ovulation in the Martina Franca donkey. Twenty-two jennies were ultrasonographically monitored and treated 72h after the detection of ovulation with 0.075 mg i.m. of R-cloprostenol. From the day of ovulation until ovulation post-treatment, blood was collected daily for P4 determination by enhanced luminescence immunoassay. All the jennies except one, exhibited behavioral signs of PGF(2alpha)-induced estrus within 4 days of treatment lasting 5.4+/-1.16 days. Post-treatment ovulation was also hastened, reducing the interovulatory interval (9.6 days). In response to treatment, plasma P4 concentrations fell to estrus levels and then remained constant until the next ovulation in all but the non-responding animal. Our findings indicate that PGF(2alpha) treatment on Day 3 post-ovulation causes the functional regression of the CL in the jenny, reflected both by the rapid induction of estrus and ovulation and by an abrupt drop in circulating P4 concentrations.
Assuntos
Corpo Lúteo/efeitos dos fármacos , Dinoprosta/farmacologia , Equidae/fisiologia , Ocitócicos/farmacologia , Animais , Dinoprosta/administração & dosagem , Estro/fisiologia , Feminino , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/efeitos dos fármacos , Ovulação , Ocitócicos/administração & dosagem , Progesterona/sangue , Fatores de TempoRESUMO
This study was designed to establish the morphological features of the placenta of the Martina Franca jenny. Ten placentas were harvested at the time of foal delivery and examined both for gross and histological characteristics. The following factors were determined: the total weight and volume of the placenta and its components, the surface area of the allantochorion, umbilical cord length and site of insertion, and the diameter of the umbilical cord vessels and vascular pattern type. The weight of the placenta was similar to previously reported for ponies, and represented 12% of foal birth weight. Umbilical cord length was similar to that in the horse but longer than in the pony, while cord weight was intermediate between the two. In a histological examination, numerous strong villi were observed at sites corresponding to the non-pregnant and pregnant horn and uterine body. No villi were detected in the area overlying the cervical star. Despite obvious similarities between the donkey and horse placenta, specific morphological features do exist, and are possibly related to the differences in length of gestation.
Assuntos
Equidae/anatomia & histologia , Placenta/anatomia & histologia , Animais , Animais Recém-Nascidos , Peso ao Nascer/fisiologia , Equidae/fisiologia , Feminino , Histocitoquímica , Masculino , Tamanho do Órgão/fisiologia , Placenta/citologia , Placenta/fisiologia , Placenta/ultraestrutura , Gravidez , Cordão Umbilical/anatomia & histologia , Cordão Umbilical/ultraestruturaAssuntos
Mutação Puntual/genética , Complicações Hematológicas na Gravidez/etiologia , Doenças de von Willebrand/genética , Fator de von Willebrand/genética , Desamino Arginina Vasopressina/administração & dosagem , Fator VIII/efeitos dos fármacos , Fator VIII/metabolismo , Feminino , Hemorragia/etiologia , Hemorragia/terapia , Heterozigoto , Humanos , Mutação de Sentido Incorreto , Fenótipo , Hemorragia Pós-Parto/tratamento farmacológico , Hemorragia Pós-Parto/etiologia , Hemorragia Pós-Parto/prevenção & controle , Gravidez , Complicações Hematológicas na Gravidez/sangue , Complicações Hematológicas na Gravidez/tratamento farmacológico , Doenças de von Willebrand/complicações , Doenças de von Willebrand/tratamento farmacológico , Fator de von Willebrand/efeitos dos fármacos , Fator de von Willebrand/metabolismoRESUMO
A phosphorylatable protein band of about 94 kDa (as judged by SDS-PAGE) which co-purifies and co-immunoprecipitates with Golgi apparatus casein kinase (G-CK) from rat lactating mammary gland has been shown by mass spectrometric sequence analysis to be identical or very similar to the glucose-regulated protein, GRP94. GRP94 is also readily phosphorylated by G-CK (K(m)=0.2 microM) at seryl sites which are different from the sites affected by casein kinase-2 (CK2) in the same protein. A study with peptide substrates would indicate that the G-CK sites in GRP94 conform to the motif S-R/K-E-X (X being different from D and E) which is not recognized by CK2.
Assuntos
Complexo de Golgi/enzimologia , Proteínas de Choque Térmico HSP70/metabolismo , Lactação , Glândulas Mamárias Animais/metabolismo , Proteínas de Membrana/metabolismo , Proteínas Quinases/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Caseína Quinase II , Caseína Quinases , Feminino , Proteínas de Choque Térmico HSP70/química , Proteínas de Choque Térmico HSP70/isolamento & purificação , Cinética , Glândulas Mamárias Animais/enzimologia , Espectrometria de Massas , Proteínas de Membrana/química , Proteínas de Membrana/isolamento & purificação , Microssomos Hepáticos , Dados de Sequência Molecular , Peso Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Fosforilação , Fosfosserina/metabolismo , Testes de Precipitina , Ligação Proteica , Proteínas Quinases/isolamento & purificação , Proteínas Serina-Treonina Quinases/metabolismo , Ratos , Especificidade por SubstratoRESUMO
The hematopoietic lineage cell-specific protein HS1 was shown to undergo a process of sequential phosphorylation both in vitro and in vivo, which is synergistically mediated by Syk and Src family protein-tyrosine kinases and essential for B cell antigen receptor-mediated apoptosis. We have now identified tyrosine 222 as the HS1 residue phosphorylated by the Src family protein kinases c-Fgr and Lyn, and we show that a truncated form of HS1 (HS1-208-401) lacking the N-terminal putative DNA binding region and the C-terminal Src homology 3 (SH3) domain is still able to undergo all the steps of sequential phosphorylation as efficiently as full-length HS1. We also show that a stable association of phospho-HS1 with c-Fgr through its SH2 domain requires previous autophosphorylation of the kinase and is prevented by subsequent phosphorylation of Tyr-222. Kinetic studies with HS1 and its truncated forms previously phosphorylated by Syk and with a peptide substrate reproducing the sequence around tyrosine 222 support the view that efficient phosphorylation of HS1 by Src family protein kinases entirely relies on TyrP-SH2 domain interaction with negligible, if any, contribution of local specificity determinants. Our data indicate that the proline-rich region of HS1 bordered by tyrosyl residues affected by Syk and Src family kinases represents a functional domain designed to undergo a process of sequential phosphorylation.