RESUMEN
BACKGROUND AND OBJECTIVES: Peach gibberellin-regulated protein (peamaclein) has recently emerged as a relevant food allergen in cypress pollen-hypersensitive patients. Objective: We investigated monosensitization to peamaclein among Italian cypress pollen-allergic patients. MATERIAL AND METHODS: A total of 835 cypress pollen-hypersensitive patients from 28 Italian allergy centers underwent a thorough work-up to determine food-allergic reactions and performed skin prick testing with a commercial peach extract containing peamaclein. IgE to rPru p 3 was measured in peach reactors, and those with negative results were enrolled as potentially monosensitized to peamaclein. IgE reactivity to rPru p 7 was evaluated using immunoblot and an experimental ImmunoCAP with rPru p 7. RESULTS: Skin prick tests were positive to peach in 163 patients (19.5%); however, 127 (77.9%) were excluded because they reacted to Pru p 3. Twenty-four patients (14.7%) corresponding to 2.8% of the entire study population) were considered potentially monosensitized to peamaclein. No geographic preference was observed. Seventeen of the 24 patients (70.8%) had a history of food allergy, mainly to peach (n=15). Additional offending foods included other Rosaceae, citrus fruits, fig, melon, tree nuts, and kiwi. On peach immunoblot, only 3 of 18 putative peamaclein-allergic patients reacted to a band at about 7 kDa; an additional 4 patients reacted at about 50-60 kDa. Ten of 18 patients (56%) had a positive result for Pru p 7 on ImmunoCAP. CONCLUSION: Allergy and sensitization to peamaclein seem rare in Italy. Most patients react to peach, although other Rosaceae fruits and several citrus fruits may also be offending foods. Peach and cypress pollen probably also share cross-reacting allergens other than peamaclein.
Asunto(s)
Cupressus , Hipersensibilidad a los Alimentos , Alérgenos/efectos adversos , Antígenos de Plantas/efectos adversos , Reacciones Cruzadas , Hipersensibilidad a los Alimentos/epidemiología , Giberelinas , Humanos , Inmunoglobulina E , Proteínas de Plantas/efectos adversos , Polen , Pruebas Cutáneas/efectos adversosRESUMEN
NMDA glutamate receptors have key roles in brain development, function and dysfunction. Regulatory roles of D-serine in NMDA receptor-mediated synaptic plasticity have been reported. Nonetheless, it is unclear whether and how neonatal deficits in NMDA-receptor-mediated neurotransmission affect adult brain functions and behavior. Likewise, the role of D-serine during development remains elusive. Here we report behavioral and electrophysiological deficits associated with the frontal cortex in Pick1 knockout mice, which show D-serine deficits in a neonatal- and forebrain-specific manner. The pathological manifestations observed in adult Pick1 mice are rescued by transient neonatal supplementation of D-serine, but not by a similar treatment in adulthood. These results indicate a role for D-serine in neurodevelopment and provide novel insights on how we interpret data of psychiatric genetics, indicating the involvement of genes associated with D-serine synthesis and degradation, as well as how we consider animal models with neonatal application of NMDA receptor antagonists.
Asunto(s)
Trastornos Mentales , Proteínas Nucleares/deficiencia , Serina/uso terapéutico , Transducción de Señal/genética , 2,3,4,5-Tetrahidro-7,8-dihidroxi-1-fenil-1H-3-benzazepina/farmacología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/genética , Factores de Edad , Animales , Proteínas Portadoras/genética , Proteínas de Ciclo Celular , Modelos Animales de Enfermedad , Agonistas de Dopamina/farmacología , Antagonistas de Aminoácidos Excitadores/uso terapéutico , Conducta Exploratoria/efectos de los fármacos , Lóbulo Frontal/patología , Aprendizaje por Laberinto/efectos de los fármacos , Trastornos Mentales/tratamiento farmacológico , Trastornos Mentales/genética , Trastornos Mentales/prevención & control , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Actividad Motora/efectos de los fármacos , Actividad Motora/genética , Neuronas/efectos de los fármacos , Proteínas Nucleares/genética , Inhibición Prepulso/efectos de los fármacos , Inhibición Prepulso/genética , Serina/metabolismo , Transducción de Señal/efectos de los fármacos , Natación/psicología , Factores de TiempoRESUMEN
An educational program, consisting in a clear explanation of the technical aspects and use of the adrenaline auto-injection devices (AAD) and in a practical test utilizing a demonstration kit was given to 350 patients from our outpatient clinic. AAD was also distributed to 50 patients formerly followed by another allergy clinic without training. At practical control test, only 10% of the untrained patients where able to correctly use the AAD versus 80% of trained subjects. Since AAD is a life-saving procedure in severe anaphylactic episodes, this attempt to improve the ability of the patient to comply with the procedure can improve the efficacy of the treatment and eventually the patient's health.
Asunto(s)
Epinefrina/administración & dosificación , Educación del Paciente como Asunto , Autoadministración/instrumentación , Humanos , Inyecciones/instrumentaciónRESUMEN
BACKGROUND: Most cases of beer allergy reported so far have been associated with hypersensitivity to the non-specific lipid transfer protein (LTP). In view of the marked differences in brewing processes we assessed IgE reactivity as well as tolerance to many different beers in an allergic patient. METHODS: A 45 year-old man hypersensitive to grass pollen, cat dander and Alternaria tenuis with a history of urticaria and dyspnoea after drinking beer and a weak skin reactivity to commercial corn extract was studied. The patient underwent SPT with 36 different brands of beer and an open challenge with those scoring negative was performed. An immunoblot analysis was carried out using 2 SPT-positive beers, 2 SPT-negative beers, and barley, wheat, and maize extracts using both patient's serum and a maize LTP-specific in-house developed polyclonal antibody from rabbit. Further, the immune reactive LTP of one beer was separated by HPLC and the chromatogram was compared to that of purified maize LTP. RESULTS: Beer SPT scored positive in 30/36 cases. The immunoblot analysis showed IgE reactivity at about 10 kDa against the two SPT-positive beers and against maize with both patient's serum and the polyclonal anti-LTP rabbit serum, whereas the two SPT-negative beers, and barley extract scored negative. The immunodetected protein co-migrated with maize LTP. CONCLUSION: In beer-allergic patients the diagnostic workup may point to the detection of some tolerated products that can be consumed risk-free.
Asunto(s)
Cerveza/efectos adversos , Proteínas Portadoras/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Animales , Cerveza/clasificación , Gatos , Femenino , Hipersensibilidad a los Alimentos/etiología , Humanos , Immunoblotting , Inmunoglobulina E/sangre , Masculino , Conejos , Zea mays/inmunologíaRESUMEN
BACKGROUND: Shrimp is a frequent cause of food allergy worldwide. Besides tropomyosin, several allergens have been described recently. OBJECTIVE: We investigated which allergens are involved in Italian shrimp-allergic adults. METHODS: Sera from 116 shrimp-allergic patients selected in 14 Italian allergy centers were studied. Skin prick tests with house dust mite (HDM) as well as measurements of IgE to Pen a 1 (shrimp tropomyosin) and whole shrimp extract were performed. All sera underwent shrimp immunoblot analysis, and inhibition experiments using HDM extract as inhibitor were carried out on some Pen a 1-negative sera. RESULTS: Immunoblots showed much variability. IgE reactivity at about 30 kDa (tropomyosin) was found in <50% of cases, and reactivity at about 67 kDa and >90 kDa was frequent. Further reactivities at 14-18, 25, 43-50, about 60 and about 80 kDa were detected. Most subjects had a history of shrimp-induced systemic symptoms irrespective of the relevant allergen protein. IgE to Pen a 1 were detected in sera from 46 (41%) patients. Skin reactivity to HDM was found in 43/61 (70%) Pen 1-negative subjects and inhibition studies showed that pre-adsorption of sera with HDM extract induced a marked weakening of the signal at >67 kDa. CONCLUSIONS: Several allergens other than tropomyosin are involved in shrimp allergy in adult Italian patients. Some hitherto not described high molecular weight allergens seem particularly relevant in this population and their cross-reactivity with HDM allergens makes them novel potential panallergens of invertebrates.
Asunto(s)
Alérgenos/inmunología , Proteínas de Artrópodos/inmunología , Hipersensibilidad a los Alimentos/epidemiología , Hipersensibilidad a los Alimentos/inmunología , Adolescente , Adulto , Anciano , Animales , Niño , Femenino , Humanos , Inmunoglobulina E/inmunología , Masculino , Persona de Mediana Edad , Peso Molecular , Prevalencia , Pruebas Cutáneas , Adulto JovenRESUMEN
BACKGROUND: The worldwide prevalence of allergic diseases such as rhinitis, asthma, and atopic dermatitis is continuously increasing, while other allergic disorders such as urticaria and angioedema are less investigated. We performed a population study evaluating the prevalence of any kind of allergic disorders. METHODS: The entire population of 7,201 inhabitants of Cotignola (Ravenna, Italy) was surveyed by a questionnaire assessing symptoms related to rhinitis, asthma, anaphylaxis, skin symptoms and insect sting allergy as well as the features of clinical presentations, diagnosis, and treatment received. RESULTS: Valid questionnaires were obtained by 6,676 inhabitants (92.7%). The sample was formed by 3,266 males and 3,495 females, the mean age was 45.6 years; 1,035 subjects (15.5%) were aged less than 18 years; 404 subjects (6%) had at least one episode of wheezing/breathlessness in their lifetime, and 243 of them (60.1%) had a diagnosis of asthma; 1,002 subjects (14.8%) had nose symptoms in their lifetime, and 375 of them (37.4%) had a diagnosis of allergic rhinitis or rhinoconjunctivitis. For other allergic manifestations, data were obtained from 5,730 subjects; of them, 178 (3.1%) had skin symptoms, 59 (1.1%) had oral symptoms, and 37 (0.6%) had anaphylaxis; 207 (3.6%) had reactions to insect stings. There were no significant differences in prevalence between Italians and immigrants. Only 51.7% of subjects with asthma, 46.5% of those with rhinitis, 22.7% of those with other allergies, but 97.1% of those with insect allergy, received treatment. CONCLUSIONS: These findings confirm recent data on epidemiology of allergic diseases in Europe, particularly in Italy, and add some details on how such diseases are managed.
Asunto(s)
Hipersensibilidad/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Italia/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Encuestas y Cuestionarios , Adulto JovenRESUMEN
A 39-years-old man afferred to our hospital for a fever lasting for more than 6 months, without abnormalities at physical examination (in particular no skin alterations); a recent laboratory and instrumental investigation was ineffective and so a fever of unknown origin (FUO) was diagnosed Since he reported an history of infantile mastocytosis (usually auto-resolving) we evaluated his serum-tryptase levels that resulted of 49 ug/L (normal value 20 ug/L), raising the doubt of the presence of an active mastocytosis. The following bone marrow evaluation showed aggregates of CD117 positive cells and a c-Kit point mutation at codon D 816V confirming the diagnosis of indolent mastocytosis.The present case confirm that FUO can be caused by an otherwise asymptomatic indolent mastocytosis, thus suggesting to include the serum-tryptase level measurement in the diagnostic approach to this pathological condition, at least in selected cases.
Asunto(s)
Fiebre de Origen Desconocido/etiología , Mastocitosis/complicaciones , Adulto , Humanos , Masculino , Proteínas Proto-Oncogénicas c-kit/análisis , Triptasas/sangreRESUMEN
BACKGROUND: Lipid transfer protein (LTP), the most frequent cause of primary food allergy in Italy, is a cross-reacting plant pan-allergen. Markers able to predict whether a patient sensitized to a certain food but not yet clinically allergic will develop allergy would be extremely helpful. OBJECTIVE: It was the aim of this study to investigate the relevance of IgE levels to some plant foods other than Rosaceae as predictors of either local or systemic allergic reaction in LTP-allergic subjects. METHODS: One hundred (40 males, 60 females , mean age 29 years) peach-allergic patients monosensitized to LTP seen at 14 Italian centres in 2009 were studied. Walnut, hazelnut, peanut, tomato, rice and/or maize allergy was ascertained by interview and confirmed by positive skin prick test. IgE levels to these foods and to rPru p 3 were measured. RESULTS: Higher levels of IgE to Pru p 3 were associated with a higher prevalence of allergy to hazelnut, peanut and walnut. For all study foods, except rice, median IgE levels in allergic subjects significantly exceeded those in tolerant subjects, though within single allergic groups, the differences between patients reporting systemic or local (oral) symptoms were not significant. Ninety-five percent cut-off IgE levels predictive of clinical allergy were established for study foods although the marked overlaps between allergic and tolerant subjects made them of limited usefulness. CONCLUSION: Specific IgE levels are only partially predictive of clinical allergy. The reasons why some individuals showing low specific IgE levels develop clinical allergy whereas others showing high IgE levels do not, despite similar exposure to the allergen, remain unclear.
Asunto(s)
Alérgenos/inmunología , Reacciones Cruzadas , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/sangre , Adulto , Antígenos de Plantas , Proteínas Portadoras/inmunología , Femenino , Alimentos/efectos adversos , Hipersensibilidad a los Alimentos/fisiopatología , Humanos , Italia , Masculino , Proteínas de Plantas , Pronóstico , Rosaceae/inmunología , Pruebas Cutáneas , Encuestas y CuestionariosRESUMEN
In literature it has been described a high risk of systemic reaction after blood-sucking Dyptera bites, like mosquitoes and horsefly, in people sensitive to hymenoptera. A 51 year old man, allergic to hymenoptera venom and with a history of i.v. reaction after Mueller, who has been treated with Vespula sp. ITS for the last 3 years, was stung by a yellow, black and green insect on the neck. Five minutes after the bite, he suffered generalized hitching and urticaria, oral cavity and lower limbs paresthesia, followed by lost of consciousness. At the Emergency Room he was successfully treated with adrenaline, intravenous antihistamines and corticosteroid. The description of the insect as well as the lack of the sting on the site suggested a wasp as the culprit. By studying one of these insect that has been captured by the patient, it turned out it wasn't a Vespula, but a horsefly, the Tabanus bovinus, which resembles Hymenoptera. Skin prick test and RAST for Tabanus confirmed the allergology diagnosis. In conclusion, also Tabanus bovines can cause systemic reaction up to anaphylactic shock.
Asunto(s)
Anafilaxia/diagnóstico , Dípteros/inmunología , Himenópteros/inmunología , Inmunización , Mordeduras y Picaduras de Insectos/diagnóstico , Administración Sublingual , Corticoesteroides/administración & dosificación , Anafilaxia/inmunología , Anafilaxia/patología , Anafilaxia/fisiopatología , Anafilaxia/terapia , Animales , Sangre/metabolismo , Diagnóstico Diferencial , Epinefrina/administración & dosificación , Antagonistas de los Receptores Histamínicos/administración & dosificación , Humanos , Mordeduras y Picaduras de Insectos/inmunología , Mordeduras y Picaduras de Insectos/patología , Mordeduras y Picaduras de Insectos/fisiopatología , Mordeduras y Picaduras de Insectos/terapia , Masculino , Persona de Mediana Edad , Prurito , Inconsciencia , UrticariaRESUMEN
BACKGROUND: The glycoprotein CD30 is expressed and released by T lymphocytes that secrete type 2 helper cytokines of (T(H)2). These molecules play a role in the pathogenesis of allergic diseases. Venom immunotherapy has proven to be very effective in hymenoptera venom allergy through a shift in cytokine production from T(H)2-type cytokines to T(H)1-type cytokines. OBJECTIVE: To evaluate the relationship between the soluble form of CD30 (sCD30) and venom immunotherapy in patients with hymenoptera venom allergy. MATERIALS AND METHODS: sCD30 levels were assayed by enzyme-linked immunosorbent assay in the sera of 61 healthy controls and 14 patients with hymenoptera venom allergy who had undergone immunotherapy before treatment and 1,3, and 12 months after treatment started. Nine patients were allergic to Apis venom, 4 to Vespula venom, and 1 to Polistes venom. RESULTS: CD30 serum levels (median, interquartile range) were significantly higher in venom-allergic patients before treatment (33.6 U/mL; 14.8-61.6) than in controls (9.7 U/mL, 1.9-21.3) (P < .000). These levels decreased progressively during treatment in all patients except 2 (P < .000). At the third month of therapy, the levels reached statistical significance in comparison with baseline. CONCLUSIONS: This study shows that sCD30 levels are significantly higher in patients with hymenoptera venom allergy and indirectly confirms a preferential T(H)2-type cytokine production in these patients. sCD30 expression decreases during immunotherapy, thus confirming the immunomodulatory role of this treatment in promoting a shift to T(H)1-type cytokines.
Asunto(s)
Venenos de Artrópodos/inmunología , Desensibilización Inmunológica , Himenópteros/inmunología , Hipersensibilidad Inmediata/inmunología , Antígeno Ki-1/sangre , Adulto , Anciano , Animales , Femenino , Humanos , Hipersensibilidad Inmediata/terapia , Mordeduras y Picaduras de Insectos/inmunología , Masculino , Persona de Mediana Edad , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/inmunología , Células Th2/metabolismoRESUMEN
BACKGROUND: Adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are a frequently reported problem due to the fact that these molecules are often used for control of pain and inflammation. Although the use of selective inhibitors of cyclooxygenase (COX) 2 helps to prevent some of these adverse reactions, they can have cardiac side effects when taken for prolonged periods. Here we report the safety and tolerability of etoricoxib, a selective COX-2 inhibitor with fewer cardiovascular effects, in patients with adverse reactions to NSAIDs. PATIENTS AND METHODS: We performed placebo-controlled oral challenge with etoricoxib in 65 patients with previous adverse reactions to NSAIDs: 13 to salicylates, 18 to arylpropionic acids, 10 to arylacetic acid, 12 to oxicam and derivates, 8 to pyrazolones, and 4 to acetaminophen (paracetamol). The reported symptoms were urticaria or angioedema in 69%, rhinitis in 3%, and 1 case of anaphylactic shock (1.5%). The challenge was done using the placebo on the first day, half dosage of etoricoxib (45 mg) on the second day, and the therapeutic dose of 90 mg on the third day. The challenge was done in the outpatient department of the hospital and the subjects were monitored for a further 4 to 6 hours after challenge. RESULTS: Oral challenge with etoricoxib was well tolerated in 97% of the patients. Only 2 systemic reactions were reported during the challenge test. CONCLUSION: Etoricoxib can be considered a safe molecule for those patients with previous adverse reactions to NSAIDs.
Asunto(s)
Antiinflamatorios no Esteroideos/efectos adversos , Inhibidores de la Ciclooxigenasa/efectos adversos , Piridinas/efectos adversos , Sulfonas/efectos adversos , Administración Oral , Ciclooxigenasa 2/metabolismo , Inhibidores de la Ciclooxigenasa/administración & dosificación , Etoricoxib , Femenino , Humanos , Masculino , Persona de Mediana Edad , Piridinas/administración & dosificación , Sulfonas/administración & dosificaciónRESUMEN
Here are two cases of two male patients of 57 and 62 years of age, already known as allergic to stinging hymenoptera venom, who after a horsefly bite have presented a serious 3-4 degree-type Mueller classification systemic reaction. The diagnosis has been carried out clinically and after an accurate environmental anamnesis and along with prick tests and RAST, further specific entomological confirm. In literature the so called wasp-mosquito-syndrome has been indicated where hyaluronidase has been referred to as the cross allergen, between the hymenoptera venom and the mosquito saliva, which likely triggers the reaction. We believe that it is also possible to take into consideration a wasp-horsefly-syndrome as well, supposing the increased risk of anaphylactic reactions to Tabanidae bites, relatively frequent in areas with animals and streams, in subjects sensitized to stinging hymenoptera. We also suggest the possibility that in these subjects some systemic reactions are due in fact to Tabanidae bites and not so much for the failure of a possible active ITS of stinging hymenoptera.
Asunto(s)
Dípteros/inmunología , Hipersensibilidad Inmediata/inmunología , Mordeduras y Picaduras de Insectos/inmunología , Avispas/inmunología , Animales , Venenos de Artrópodos/efectos adversos , Venenos de Artrópodos/inmunología , Humanos , Hipersensibilidad Inmediata/etiología , Masculino , Persona de Mediana Edad , Prueba de Radioalergoadsorción , Pruebas Cutáneas , SíndromeRESUMEN
Seminoma cell lines, essential to the study of the biology of seminoma, do not exist. Tissue culture conditions for establishing such cell lines have to be developed. Under conventional culture conditions, seminoma cells usually die within the first 3 days after plating. The enhanced survival of rat gonocytes when cocultured with rat Sertoli cells in serum-free medium suggests that seminoma cells, the neoplastic counterparts of gonocytes, might benefit from the same conditions. Indeed, when cocultured with rat Sertoli cells in a serum-free medium, viable seminoma cells could be demonstrated on the 11th day of culture. This result is a significant improvement over the results with conventional methods.
Asunto(s)
Medio de Cultivo Libre de Suero/farmacología , Disgerminoma/patología , Células de Sertoli/citología , Neoplasias Testiculares/patología , Adulto , Fosfatasa Alcalina/metabolismo , Supervivencia Celular/fisiología , Disgerminoma/enzimología , Humanos , Inmunohistoquímica , Masculino , Neoplasias Testiculares/enzimología , Células Tumorales CultivadasRESUMEN
The case of a 48-year-old woman in whom focal nodular hyperplasia of the liver developed after busulfan therapy was administered for essential thrombocytosis is described. Focal nodular hyperplasia is a reactive disorder related to a circulation disorder. The close temporal relation between the haematological disease, busulfan treatment and the appearance of hyperplastic diseases of the liver in our patient supports the possibility that the association of the events might not be casual.
Asunto(s)
Busulfano/efectos adversos , Hiperplasia Nodular Focal/inducido químicamente , Inmunosupresores/efectos adversos , Busulfano/administración & dosificación , Femenino , Hiperplasia Nodular Focal/diagnóstico por imagen , Humanos , Inmunosupresores/administración & dosificación , Persona de Mediana Edad , Trombocitosis/tratamiento farmacológico , UltrasonografíaRESUMEN
Anaphylaxis is a severe, life-threatening allergic reaction, affecting both children and adults. The occurrence of anaphylaxis is not as rare as generally believed (1.21% to 15.04% of the US population). Often the cause of this reaction remain unknown, mainly due to the difficulty in defining the outbreaking causes. Herein, we describe an interesting case of a patient, who developed an anaphylactic reaction after the bite of a pigeon tick. During the last 2 years, in wintertime, the patient often came to the emergency room for general rash and swelling, hypotension and tachycardia preceded by itching and general distress. Notably, the symptoms manifested themselves as night fell. In two particular occasions the patient reached the hospital in a state of shock. After another episode of general swelling, the patient was invited to examine her domestic environment. She brought us some parasites, collected at home, particularly on the bed. A morphological examination by entomologists proved these parasites to belong to Argas reflexus (Arg.r.), one of the 31 species of soft ticks. The presence of specific IgE to a protein secreted by the Arg.r. salivary glands was in favour of immediate-type systemic reaction, as supposed by the clinical history.
Asunto(s)
Anafilaxia/etiología , Argas , Mordeduras y Picaduras/inmunología , Anafilaxia/sangre , Anafilaxia/inmunología , Animales , Argas/inmunología , Argas/fisiología , Ropa de Cama y Ropa Blanca , Enfermedades de las Aves/parasitología , Columbidae/parasitología , Femenino , Vivienda , Humanos , Inmunoglobulina E/inmunología , Persona de Mediana Edad , Recurrencia , Proteínas y Péptidos Salivales/inmunología , Infestaciones por Garrapatas/parasitología , Infestaciones por Garrapatas/veterinariaRESUMEN
Bovine follicular fluid (bFF) injected ip in mice during 2 days (65,000 U inhibin/day, 1 U inhibin the activity in 1 micrograms bFF protein) caused a significant decrease in the numbers of A4, intermediate (In), and B spermatogonia to 91%, 74%, and 67% of the control values, respectively. The numbers of undifferentiated spermatogonia remained unchanged. These injections suppressed peripheral FSH levels to 6% of the control values, suggesting that FSH might be the modulator of the effects on spermatogenesis. However, in the Chinese hamster, intratesticular injections of bFF during 4 days (6500 U inhibin/day into one testis) also caused a significant decrease in the numbers of A3. In, B1, and B2 spermatogonia to 86%, 61%, 55%, and 94% of the control values, respectively. Similarly, treatment with a partially purified inhibin preparation from rat Sertoli cell-conditioned medium (rSCCM) during 4 days (Mono Q fraction; 1512 U inhibin/day; 37.8 micrograms protein) caused a significant decrease in the numbers of A3, In, B1, and B2 spermatogonia to 90%, 87%, 66%, and 93% of the control values, respectively. Treatment with a highly purified inhibin preparation from rSCCM during 4 days (30K inhibin; 750 U inhibin/day; 100 ng protein) significantly decreased the numbers of In and B1 spermatogonia to, respectively, 87% and 91% of the control values. These effects were limited to the testis into which the material was injected; the contralateral testis or testes injected with control fluid always showed normal numbers of spermatogonia. This implies that the effects on the seminiferous epithelium are not FSH mediated. Intratesticular injections of bFF or pure inhibin did not affect the number of undifferentiated spermatogonia. However, the Mono Q fraction caused a significant increase in the numbers of undifferentiated spermatogonia in stages IV-VII of the cycle, suggesting the presence of a mitogenic factor for undifferentiated spermatogonia in rSCCM which is not present or is counteracted in bFF. The results suggest that inhibin may have a role in the regulation of spermatogonial development in the adult animal.
Asunto(s)
Inhibinas/farmacología , Espermatogonias/citología , Testículo/citología , Animales , Bovinos , Recuento de Células/efectos de los fármacos , Cricetinae , Medios de Cultivo , Femenino , Líquido Folicular/fisiología , Inhibinas/metabolismo , Inyecciones , Inyecciones Intraperitoneales , Masculino , Mesocricetus , Ratones , Ratones Endogámicos , Peso Molecular , Células de Sertoli/citología , Células de Sertoli/metabolismo , EspermatozoidesRESUMEN
Due to the lack of a specific marker for gonocytes from newborn rats, isolation of these cells has proven difficult and laborious. We have found a specific cell membrane marker, the Epithelial Cellular Adhesion Molecular (Ep-CAM) that can be used to isolate these cells using antibody directed cell sorting. 4 days post partum (dpp) rat testes were enzyme treated to attain a cell suspension, which was labelled with an antibody (GZ1) against Ep-CAM and tagged with a fluorescent probe. The labelled cell suspension was run through a FACS cell sorter, from which a gonocyte suspension of >85% purity was attained. The cells remained viable in culture and proliferated actively as determined by double labelling the cells with anti-HSP90alpha (a specific germ cell marker) and anti-BrdU antibodies (after BrdU incorporation). During culture, these cells formed chains of 2 to 4 cells and aggregates of proliferating germ cells were found after 8 days of culture.
Asunto(s)
Antígenos de Neoplasias/análisis , Moléculas de Adhesión Celular/análisis , Separación Celular/métodos , Espermatozoides/fisiología , Animales , Animales Recién Nacidos , División Celular , Células Cultivadas , Molécula de Adhesión Celular Epitelial , Citometría de Flujo , Masculino , Ratas , Ratas WistarRESUMEN
Sertoli cell-gonocyte cocultures obtained from rat testes 20 days postcoitum, 1 day postpartum, and 3 days postpartum were used to investigate the effect of FGF-2 on both somatic and germ cells in vitro during the perinatal period. With cells isolated from fetal, newborn, or 3-day-old animals, FGF-2 was found to significantly increase the number of Sertoli cells after 3 or 6 days of cultures, starting at a concentration of 1 ng/ml. FGF-2 did not increase the [3H]thymidine labeling index of Sertoli cells, indicating that FGF-2 is a survival factor for these cells in vitro. FGF-2 (1, 5, or 10 ng/ml) also significantly increased the number of gonocytes after 6 days of culture with cells from either newborn or 3-day-old animals. About twice as many germ cells were found in those cultures compared to the control cultures. Addition of a neutralizing antibody against FGF-2 to control cultures caused a significant decrease in the number of gonocytes compared to that in untreated cultures after 6 days, whereas with FGF-2, the antibody decreased the number of germ cells to control levels. FGF-2 significantly stimulated the proliferative activity of the gonocytes after 3 or 5 days, indicating that FGF-2 is a survival as well as a mitogenic factor for these cells. Taken together, these data suggest that FGF-2 is an important factor around the start of spermatogenesis, at least in vitro.
Asunto(s)
Animales Recién Nacidos/fisiología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Células de Sertoli/efectos de los fármacos , Testículo/citología , Testículo/efectos de los fármacos , Envejecimiento/fisiología , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Recuento de Células , División Celular/efectos de los fármacos , Supervivencia Celular , Técnicas de Cocultivo , Factor 2 de Crecimiento de Fibroblastos/antagonistas & inhibidores , Gónadas/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Células de Sertoli/citologíaRESUMEN
Leukemia inhibitory factor (LIF) and ciliary neurotropic factor (CNTF) were found to be pleiotropic modulators of Sertoli cell and gonocyte development (both isolated from the neonatal rat testis) in a coculture system, whereas IL-6, another member of this cytokine family, had no effect on these cells. LIF and CNTF significantly enhanced the survival of the Sertoli cells in a dose- and time-dependent manner. The effect of LIF on the Sertoli cells was significant at a concentration of 1 ng/ml after 3 or 6 days of culture, whereas CNTF had a significant effect at 10 ng/ml. Neither LIF nor CNTF had an effect on Sertoli cell proliferation. The survival of proliferating gonocytes (isolated from 3-day-old rats testes) was also significantly higher in cultures to which LIF (7.5 ng/ml) or CNTF (10 ng/ml) was added. No effect of these cytokines was found on the mitotic activity of proliferating gonocytes. However, LIF (7.5 ng/ml) stimulated the proliferation of quiescent gonocytes (isolated from day 1 testes) after 3 days of culture. Combinations of LIF (or CNTF) with fibroblast growth factor 2 (10 ng/ml) and steel factor (50 ng/ml) did not further improve the long term culture of the gonocytes. LIf- and CNTF-like proteins of the expected molecular masses (32,000 and 22,000 daltons, respectively, under reducing conditions) were found by Western blotting in testicular extracts of 3-day-old rats. Taken together, these results indicate that LIF or CNTF may play a role at the start of the spermatogenesis. The characterization of receptors for LIF or CNTF on the gonocytes and/or neonatal Sertoli cells will aid in a better understanding of the physiological role of these cytokines in the reproductive system.