RESUMO
The endoplasmic reticulum (ER) is the start site of the secretory pathway, where newly synthesized secreted and membrane proteins are packaged into COPII vesicles through direct interaction with the COPII coat or aided by specific cargo receptors. Little is known about how post-translational modification events regulate packaging of cargo into COPII vesicles. The Saccharomyces cerevisiae protein Erv14, also known as cornichon, belongs to a conserved family of cargo receptors required for the selection and ER export of transmembrane proteins. In this work, we show the importance of a phosphorylation consensus site (S134) at the C-terminus of Erv14. Mimicking phosphorylation of S134 (S134D) prevents the incorporation of Erv14 into COPII vesicles, delays cell growth, exacerbates growth of sec mutants, modifies ER structure and affects localization of several plasma membrane transporters. In contrast, the dephosphorylated mimic (S134A) had less deleterious effects, but still modifies ER structure and slows cell growth. Our results suggest that a possible cycle of phosphorylation and dephosphorylation is important for the correct functioning of Erv14.
Assuntos
Proteínas de Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Membrana/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Transporte Biológico , Vesículas Revestidas pelo Complexo de Proteína do Envoltório/metabolismo , Transporte ProteicoRESUMO
Variable domain (VL) gene segments exhibit variable tendencies to be associated with light chain amyloidosis (AL). While few of them are very frequent in AL and give rise to most of the amyloidogenic light chains compiled at the sequence databases, other are rarely found among the AL cases. To analyze to which extent these tendencies depend on folding stability and aggregation propensity of the germline VL protein, we characterized VL proteins encoded by four AL-associated germline gene segments and one not associated to AL. We found that the AL-associated germline rVL proteins differ widely in conformational stability and propensity to in vitro amyloid aggregation. While in vitro the amyloid formation kinetics of these proteins correlate well with their folding stabilities, the folding stability does not clearly correlate with their germline's frequencies in AL. We conclude that the association of the VL genes segments to amyloidosis is not determined solely by the folding stability and aggregation propensity of the germline VL protein. Other factors, such as the frequencies of destabilizing mutations and susceptibility to proteolysis, must play a role in determining the light chain amyloidogenicity.
Assuntos
Amiloide/genética , Amiloidose/genética , Região Variável de Imunoglobulina/genética , Agregação Patológica de Proteínas/genética , Sequência de Aminoácidos , Mutação em Linhagem Germinativa , Humanos , Microscopia Eletrônica de Transmissão , Domínios Proteicos , Estabilidade Proteica , Alinhamento de Sequência , Espectrometria de FluorescênciaRESUMO
The study of sexual reproductive behavior supported by ultrastructural evidence is important in rotifers to describe differences among potential cryptic species. In this research, the morphology of the rotifer Brachionus bidentatus is described at the ultrastructural level, using electronic microscopy, together with a brief description and discussion of its sexual reproductive behavior. The characteristics of the (a) male, (b) the female, (c) the sexual egg or cyst, (d) the partenogenic egg, (e) the no-fecundated sexual egg (male egg), and (f) the trophi, were described. Another part of this research is dedicated to the ultrastructure of the sex cells of the male rotifer B. bidentatus. Samples were obtained from La Punta pond in Cosio, Aguascalientes, Mexico (22 degrees 08' N - 102 degrees 24' W), and a culture was maintained in the laboratory. Fifty organisms, from different stages of the rotifer Brachionus bidentatus, were fixed in Formol at 4% and then prepared; besides, for the trophi, 25 female rotifer Brachionus bidentatus were prepared for observation in a JEOL 5900 LV scanning electronic microscope. In addition, for the observation of male sex cells, 500 males of Brachionus bidentatus were isolated, fixed and observed in a JEOL 1010 transmission microscope. Females of B. bidentatus in laboratory cultures had a lifespan of five days (mean+one SD = 4.69 +/- 0.48; N=13), and produced 4.5 +/- 3.67 (N=6) parthenogenetic eggs during such lifespan. In the case of non-fertilized sexual eggs, they produced up to 18 eggs (mean+one SD = 13 +/- 4.93; N=7). Sexual females produced a single cyst on average (mean +/- one SD = I +/- 0; N=20). For the sexual cycle, the time of copulation between male and female ranged from 10 to 40 seconds (mean +/- one SD = 17.33 +/- 10.55, N=7). The spermatozoa are composed of a celular body and a flagellum, the size of the body is of 300 nm while the flagellum measures 1 700nm. The rods have a double membrane. Their mean length is almost 2.45 microm +/- 0.74, N=6; and their mean wide is 0.773 microm +/- 0.241, N=11. The evidence on the specific ultrastructural characteristics of the rotifer B. bidentatus is notorious, even more in the male and in the cyst cell. Regarding the ultrastructure of the spermatozoa and the rods, compared to other species they only differ in size, despite their structural resemblance. Our study of the ultraestructure of this species adds useful information that along with molecular data will help clarify the taxonomy of brachionid rotifers.
Assuntos
Óvulo/ultraestrutura , Rotíferos/ultraestrutura , Espermatozoides/diagnóstico por imagem , Animais , Feminino , Masculino , México , Microscopia Eletrônica de Transmissão , Reprodução , Rotíferos/anatomia & histologia , UltrassonografiaRESUMO
OBJECTIVE: To report a case of a woman who used duloxetine during pregnancy and breast-feeding. CASE SUMMARY: A 29-year-old woman was treated with duloxetine for depression during the second half of an uncomplicated gestation. She gave birth at term to a healthy female infant. A cord blood sample was obtained at birth. The mother continued the antidepressant while exclusively breast-feeding her infant. One month later, we collected blood and milk samples from the mother and a single blood sample from the infant. All samples were analyzed for the presence and concentrations of duloxetine. DISCUSSION: Duloxetine crosses the placenta at term and is excreted into breast milk. No evidence of developmental or other type of toxicity was observed in the infant at birth or during the first 32 days after birth. The published literature detailing human pregnancy experience with this antidepressant is limited to 11 cases in which women became pregnant while taking duloxetine. In 10 cases, the drug was discontinued when pregnancy was diagnosed and no outcome data were reported. In the eleventh case, an infant exposed to duloxetine 90 mg/day developed neonatal behavioral syndrome. One study examined the excretion of duloxetine into breast milk, but the mothers discontinued nursing for the study. In the present case, no adverse effects from exposure to the drug in milk were noted in the exclusively breast-fed infant. The possibility of functional/neurobehavioral deficits appearing later in life cannot be excluded because long-term follow-up has not been conducted in infants exposed to duloxetine in utero or during nursing. CONCLUSIONS: No developmental toxicity or other signs of toxicity were observed in an infant exposed to duloxetine during the second half of gestation and during breast-feeding in the first 32 days after birth. However, the possibility of functional/neurobehavioral deficits appearing later in life cannot be excluded.
Assuntos
Lactação/sangue , Troca Materno-Fetal/fisiologia , Leite Humano/metabolismo , Tiofenos/sangue , Adulto , Cloridrato de Duloxetina , Feminino , Humanos , Lactente , Lactação/efeitos dos fármacos , Troca Materno-Fetal/efeitos dos fármacos , Leite Humano/química , Leite Humano/efeitos dos fármacos , Gravidez , Tiofenos/uso terapêuticoRESUMO
Inulin has interesting physicochemical and functional properties, and therefore a wide range of applications in the food and medical industries. It has gained great traction due to its ability to form nanoparticles and its possible application as nanovehicle for drug delivery. In this work, we demonstrated that the enzymatically-synthesized high molecular weight (HMW) inulin forms stable spherical nanoparticles with an average diameter of 112 ± 5 nm. The self-assemblage of HMW inulin nanoparticles is carried out during enzymatic synthesis of the polymer, and become detectable after a certain critical aggregation concentration (CAC) is reached. Both, the CAC and nanoparticle size are influenced by the reaction temperature. These nanoparticles are not toxic for peripheral blood mononuclear cells, at concentrations below 200 µg/mL; no significant prebiotic potential was detected in cultures of 13 probiotic strains. This work contributes to a better understanding of the formation of HMW inulin nanoparticles and their biological properties.
Assuntos
Portadores de Fármacos/síntese química , Portadores de Fármacos/toxicidade , Hexosiltransferases/química , Inulina/síntese química , Inulina/toxicidade , Leuconostoc/enzimologia , Nanopartículas/química , Nanopartículas/toxicidade , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Feminino , Humanos , Inulina/química , Leucócitos Mononucleares/efeitos dos fármacos , Peso Molecular , Prebióticos , ProbióticosRESUMO
Immunoglobulin light chain-derived (AL) amyloidosis is a debilitating disease without known cure. Almost nothing is known about the structural factors driving the amyloidogenesis of the light chains. This study aimed to identify the fibrillogenic hotspots of the model protein 6aJL2 and in pursuing this goal, two complementary approaches were applied. One of them was based on several web-based computational tools optimized to predict fibrillogenic/aggregation-prone sequences based on different structural and biophysical properties of the polypeptide chain. Then, the predictions were confirmed with an ad-hoc synthetic peptide library. In the second approach, 6aJL2 protein was proteolyzed with trypsin, and the products incubated in aggregation-promoting conditions. Then, the aggregation-prone fragments were identified by combining standard proteomic methods, and the results validated with a set of synthetic peptides with the sequence of the tryptic fragments. Both strategies coincided to identify a fibrillogenic hotspot located at the CDR1 and ß-strand C of the protein, which was confirmed by scanning proline mutagenesis analysis. However, only the proteolysis-based strategy revealed additional fibrillogenic hotspots in two other regions of the protein. It was shown that a fibrillogenic hotspot associated to the CDR1 is also encoded by several κ and λ germline variable domain gene segments. Some parts of this study have been included in the chapter "The Structural Determinants of the Immunoglobulin Light Chain Amyloid Aggregation", published in Physical Biology of Proteins and Peptides, Springer 2015 (ISBN 978-3-319-21687-4).
Assuntos
Amiloide/metabolismo , Regiões Determinantes de Complementaridade , Cadeias Leves de Imunoglobulina/metabolismo , Amiloidose de Cadeia Leve de Imunoglobulina/metabolismo , Agregação Patológica de Proteínas/metabolismo , Sequência de Aminoácidos , Amiloide/química , Humanos , Cadeias Leves de Imunoglobulina/química , Modelos Moleculares , Conformação Proteica em Folha beta , Multimerização ProteicaRESUMO
OBJECTIVE: To report a case of use of high-dose carisoprodol during pregnancy and breast-feeding. CASE SUMMARY: A 28-year-old woman with severe back muscle spasm took carisoprodol 2800 mg/day before and throughout an uncomplicated pregnancy and while exclusively breast-feeding her infant during the first month after birth. Serum drug concentrations of carisoprodol and the active metabolite meprobamate were measured in the mother and infant. Concentrations of these agents also were measured in breast milk. Developmental toxicity was not observed in the near-term infant, whose birth weight was at the 10th percentile for gestational age. Only slight sedation was noted in the infant during breast-feeding, and no signs or symptoms of withdrawal were noted when nursing was stopped. DISCUSSION: Carisoprodol and meprobamate are excreted into breast milk. Although the published human pregnancy data are limited to 15 cases, carisoprodol does not appear to cause developmental toxicity (growth restriction, structural anomalies, functional/neurobehavioral deficits, or death), even when the mother is taking high doses. No signs or symptoms of withdrawal were noted in our infant or in a previously published case when breast-feeding was stopped. Long-term follow-up has not been conducted in exposed infants, and the possibility of functional/neurobehavioral l deficits appearing later in life cannot be excluded. CONCLUSIONS: Except for mild sedation, no other toxicity was observed in a near-term infant exposed to carisoprodol throughout gestation and during breast-feeding in the first month after birth.
Assuntos
Dor nas Costas/tratamento farmacológico , Carisoprodol/uso terapêutico , Relaxantes Musculares Centrais/uso terapêutico , Complicações na Gravidez/tratamento farmacológico , Adulto , Transporte Biológico , Aleitamento Materno , Carisoprodol/efeitos adversos , Carisoprodol/farmacocinética , Feminino , Humanos , Recém-Nascido , Troca Materno-Fetal , Meprobamato/farmacocinética , Leite Humano/metabolismo , Relaxantes Musculares Centrais/efeitos adversos , Relaxantes Musculares Centrais/farmacocinética , Gravidez , Espasmo/tratamento farmacológicoRESUMO
OBJECTIVE: To determine if glyburide and glipizide are excreted into breast milk and if breast-feeding from women taking these drugs causes infant hypoglycemia. RESEARCH DESIGN AND METHODS: We studied eight women who had received a single oral dose of 5 or 10 mg glyburide. Drug concentrations were measured in maternal blood and milk for 8 h after the dose. In a separate study, five women were given a daily dosage (5 mg/day) of glyburide or glipizide, starting on the first postpartum day. Maternal blood and milk drug concentrations and infant blood glucose were measured 5-16 days after delivery. RESULTS: In the single-dose glyburide study, the mean maximum theoretical infant dose (MTID) as a percent of the weight-adjusted maternal dose (WAMD) was <1.5 and <0.7% for the 5- and 10-mg doses, respectively. For the five women taking daily dosages, the mean MTID as a percent of the WAMD was <28% for glyburide and <27% for glipizide. The high estimates were due to the insensitivity of the assay. Neither glyburide nor glipizide were detected in breast milk in either study and blood glucose was normal in the three infants (one glyburide and two glipizide) who were wholly breast-fed when the drug concentrations were at steady state. CONCLUSIONS: Neither glyburide nor glipizide were detected in breast milk, and hypoglycemia was not observed in the three nursing infants. Both agents, at the doses tested, appear to be compatible with breast-feeding.
Assuntos
Glipizida/farmacocinética , Glibureto/farmacocinética , Hipoglicemiantes/metabolismo , Leite Humano/química , Adulto , Feminino , Glipizida/sangue , Glibureto/sangue , Humanos , Lactente , Alimentos Infantis/análiseRESUMO
BACKGROUND: Allergic sensitization is dependent on the geographical and climatic conditions in each region; therefore, identify agents most commonly sensitized children with asthma is important for planning prevention measures. OBJECTIVE: To determine the prevalence of the sensitization to allergens in children with asthma. MATERIAL AND METHOD: This cross-sectional and prolective study includes children from ages 6 to 14, who have asthma, attended for the first time in an allergy service. The patients were recruited consecutively between the months of January 1st 2014 to December 31st 2014. The frequency of the allergen sensitization was estimated. RESULTS: This study included 186 children, the median age was 7 years olds, the male group was 104/186 (55.9%) The median of the positive results was 5 and monosensitized were 47/186 (2.2%). The most common category of allergens was the indoor (90.3%), then trees (71.0%), and finally the fungi (9.7%). Individually, the house dust mites were more common in between the interior allergens, followed by the epithelial; in the tree pollen were oaks (31.7%) and ashes (28.0%), in weeds was mugwort (21.5%), in grasses was Zea mays (18.3%) and in the fungi was Cladosporium spp. (6.5%). CONCLUSION: The high prevalence of the sensitization to house dust mites and epithelial in children with asthma, stimulates to implement methods of environmental control for contributing a better control of the disease.
Antecedentes: la prevalencia de rinosinusitis crónica en pacientes adultos con inmunodeficiencia común variable (IDCV) es de 52%. Los pacientes con esta enfermedad tienen mayor frecuencia de rinosinusitis crónica, enfermedad inflamatoria que afecta a la mucosa de uno o más senos paranasales y la cavidad nasal. Objetivo: identificar los microorganismos de secreción del meato medio obtenida por endoscopia asociados con rinosinusitis crónica en pacientes adultos con inmunodeficiencia común variable (IDCV). Material y método: estudio descriptivo, transversal, que incluyó a pacientes adultos con inmunodeficiencia común variable, de quienes se obtuvo una muestra vía endoscópica de secreción del meato medio de ambas fosas nasales, que se envió a cultivo para bacterias aerobias, anaerobias y hongos. Se obtuvo consentimiento informado de todos los pacientes. Resultados: se estudiaron 29 pacientes: 18 mujeres y 11 hombres, con edad promedio de 40±13 años. Los resultados obtenidos fueron: 2 muestras de pacientes no tuvieron desarrollo microbiano, 24 tuvieron desarrollo de bacterias aerobias, en 3 casos hubo crecimiento fúngico sin desarrollo de bacterias anaerobias. Conclusiones: nuestros resultados muestran que los microorganismos asociados con rinosinusitis crónica en pacientes adultos con inmunodeficiencia común variable más comunes son: Moraxella catarrhalis, Staphylococcus, Sphingomonas paucimobilis y Citrobacter koseri; los agentes micóticos asociados fueron: Candida albicans y Aspergillus fumigatus.
Assuntos
Alérgenos/imunologia , Asma/epidemiologia , Adolescente , Animais , Asma/imunologia , Criança , Cidades/epidemiologia , Cladosporium/imunologia , Estudos Transversais , Feminino , Humanos , Masculino , México/epidemiologia , Plantas Daninhas/imunologia , Pólen/imunologia , Prevalência , Pyroglyphidae/imunologia , Testes Cutâneos , Zea mays/imunologiaRESUMO
Studies suggest that children who start solid foods early are at risk for developing food allergies. Herein, we evaluated the effects of the introduction of peanuts to the diets of children on emerging peanut allergies. Children with allergic rhinitis and asthma were enrolled in the present study and evaluated in four stages. In the first stage, a clinical history was completed for all participants. In the second stage, skin tests were conducted to detect the sensitization to peanuts. In the third stage, the parents were interviewed about the peanut-eating habits of their children. In the fourth stage, children with a convincing history of allergy or a positive peanut skin test result were subjected to an open oral food challenge (OOFC). Three hundred children in four groups were included, 58.2% of the subjects were male, and the mean age was 7.3±3.9 years. The median age of first exposure to peanuts in patients with peanut allergies was greater than that in children without peanut allergies (2 years versus 1 year; p=0.009). The multivariate analysis, including only those children subjected to the OOFC, revealed that the consumption of peanuts after the age of ≥2 years is a risk factor for developing a peanut allergy (odds ratio=8.0, 95% confidence interval 1.3-50.0, p=0.026). The results of the present study showed that the late introduction of peanuts to children increases the risk of developing a peanut allergy.
Assuntos
Arachis/efeitos adversos , Asma/epidemiologia , Dermatite Atópica/epidemiologia , Dieta/efeitos adversos , Hipersensibilidade a Amendoim/epidemiologia , Rinite Alérgica/epidemiologia , Fatores Etários , Arachis/imunologia , Asma/diagnóstico , Asma/imunologia , Asma/prevenção & controle , Criança , Pré-Escolar , Estudos Transversais , Dermatite Atópica/diagnóstico , Dermatite Atópica/imunologia , Dermatite Atópica/prevenção & controle , Feminino , Humanos , Lactente , Modelos Logísticos , Masculino , México/epidemiologia , Análise Multivariada , Razão de Chances , Hipersensibilidade a Amendoim/diagnóstico , Hipersensibilidade a Amendoim/imunologia , Hipersensibilidade a Amendoim/prevenção & controle , Prevalência , Fatores de Proteção , Rinite Alérgica/diagnóstico , Rinite Alérgica/imunologia , Rinite Alérgica/prevenção & controle , Fatores de Risco , Testes CutâneosRESUMO
BACKGROUND: Ralstonia pickettii is a Gram-negative bacillus commonly found in soil and moist environments; however, R. pickettii is rarely isolated from clinical specimens. In August 2001, a cluster of R. pickettii bacteremia occurred among neonatal intensive care unit (NICU) infants at a California hospital. METHODS: A case-control study was conducted to determine risk factors for infection. A case was a NICU patient with R. pickettii bacteremia. Controls were NICU infants with negative blood cultures drawn during the same time period. A detailed environmental investigation was also conducted. RESULTS: We identified 18 patients with 19 distinct episodes of R. pickettii bacteremia from July 30 through August 30, 2001. All cases had intravascular access at the time of bacteremia. Although the case-control study did not implicate any statistically significant risk factors, the most likely source of the outbreak was the heparin flush prepared in the hospital pharmacy. This is supported by the following: (1) the heparin flush was the only substance introduced directly into the bloodstream of all case infants; (2) the heparin flush was used exclusively by the NICU; and (3) no further cases were identified after the heparin flush was discontinued. Cultures of remaining heparin flush and environmental cultures from the NICU were negative for R. pickettii. CONCLUSIONS: This unusual outbreak of R. pickettii bacteremia was most likely caused by contaminated heparin flush and ended after the heparin flush was discontinued.
Assuntos
Bacteriemia/epidemiologia , Infecção Hospitalar/epidemiologia , Surtos de Doenças , Infecções por Bactérias Gram-Negativas/epidemiologia , Doenças do Prematuro/epidemiologia , Unidades de Terapia Intensiva Neonatal , Ralstonia pickettii/isolamento & purificação , Anticoagulantes/administração & dosagem , Bacteriemia/microbiologia , Estudos de Casos e Controles , Cateterismo Venoso Central , Infecção Hospitalar/microbiologia , Contaminação de Medicamentos , Feminino , Infecções por Bactérias Gram-Negativas/microbiologia , Heparina/administração & dosagem , Humanos , Lactente , Recém-Nascido de Baixo Peso , Recém-Nascido , Recém-Nascido Prematuro , Doenças do Prematuro/microbiologia , MasculinoRESUMO
OBJECTIVE: To determine whether metformin is excreted into breast milk and whether this exposure adversely affects the blood glucose of nursing infants. METHODS: Seven women were started on metformin 500 mg twice daily on the first day after cesarean delivery. Breastfeeding was started at the same time. Two women were excluded. Two other women stopped breastfeeding for personal reasons unrelated to the drug therapy, but did provide serum and milk samples, because they regularly pumped their breasts to maintain lactation. Peak and trough serum and milk samples were drawn between postoperative days 4 and 17. In 3 infants, blood was drawn for glucose determination at the same time as the maternal samples. RESULTS: The trough milk concentration in 1 subject was below the assay detection limit. Excluding this subject, the mean peak and trough serum metformin concentrations were 1.06 mug/mL (range 0.68-1.90 mug/mL) and 0.42 mug/mL (range 0.26-0.51 mug/mL), respectively, whereas the mean peak and trough metformin concentrations in breast milk were 0.42 mug/mL (range 0.38-0.46 mug/mL) and 0.39 mug/mL (range 0.31-0.52 mug/mL), respectively. The mean milk:serum ratio was 0.63 (range 0.36-1.00) and the mean estimated infant dose as a percentage of the mother's weight-adjusted dose was 0.65% (range 0.43-1.08%). In 3 infants, the blood glucose concentrations 4 hours after a feeding were within the normal limit, ranging from 47-77 mg/dL. CONCLUSION: Metformin is excreted into breast milk, but the amounts seem to be clinically insignificant. No adverse effects on the blood glucose of the 3 nursing infants were measured.
Assuntos
Glicemia/análise , Aleitamento Materno , Hipoglicemiantes/farmacocinética , Metformina/farmacocinética , Leite Humano/química , Adulto , Cesárea , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/sangue , Lactente , Recém-Nascido/sangue , Metformina/administração & dosagem , Metformina/sangue , Estudos ProspectivosRESUMO
The study of sexual reproductive behavior supported by ultrastructural evidence is important in rotifers to describe differences among potential cryptic species. In this research, the morphology of the rotifer Brachionus bidentatus is described at the ultrastructural level, using electronic microscopy, together with a brief description and discussion of its sexual reproductive behavior. The characteristics of the (a) male,(b) the female, (c) the sexual egg or cyst, (d) the partenogenic egg, (e) the no-fecundated sexual egg (male egg), and (f) the trophi, were described. Another part of this research is dedicated to the ultrastructure of the sex cells of the male rotifer B. bidentatus. Samples were obtained from La Punta pond in Cosio, Aguascalientes, Mexico (22°08’ N - 102°24’ W), and a culture was maintained in the laboratory. Fifty organisms, from different stages of the rotifer Brachionus bidentatus, were fixed in Formol at 4% and then prepared; besides, for the trophi, 25 female rotifer Brachionus bidentatus were prepared for observation in a JEOL 5900 LV scanning electronic microscope. In addition, for the observation of male sex cells, 500 males of Brachionus bidentatus were isolated, fixed and observed in a JEOL 1010 transmission microscope. Females of B. bidentatus in laboratory cultures had a lifespan of five days (mean±one SD=4.69±0.48; N=13), and produced 4.5+3.67 (N=6) parthenogenetic eggs during such lifespan. In the case of non-fertilized sexual eggs, they produced up to 18 eggs (mean±one SD=13±4.93; N=7). Sexual females produced a single cyst on average (mean±one SD=1±0; N=20). For the sexual cycle, the time of copulation between male and female ranged from 10 to 40 seconds (mean±one SD=17.33±10.55, N=7). The spermatozoa are composed of a celular body and a flagellum, the size of the body is of 300nm while the flagellum measures 1 700nm. The rods have a double membrane. Their mean length is almost 2.45µm±0.74, N=6; and their mean wide is 0.773µm±0.241, N=11. The evidence on the specific ultrastructural characteristics of the rotifer B. bidentatus is notorious, even more in the male and in the cyst cell. Regarding the ultrastructure of the spermatozoa and the rods, compared to other species they only differ in size, despite their structural resemblance. Our study of the ultraestructure of this species adds useful information that along with molecular data will help clarify the taxonomy of brachionid rotifers. Rev. Biol. Trop. 61 (4): 1737-1745. Epub 2013 December 01.
El estudio del comportamiento reproductivo sexual apoyado en evidencias ultraestructurales en rotíferos, es importante para describir diferencias entre especies potencialmente crípticas. En este trabajo se describe a nivel ultraestructural la morfología del rotífero Brachionus bidentatus, usando microscopía electrónica, junto con una breve descripción y discusión de su comportamiento sexual reproductivo. Se presentan las características del: (a) macho, (b) hembra, (c) huevo partenogenético, (d) huevo sexual no fecundado, (e) trofos. También se muestra un apartado sobre la ultra estructura de las células sexuales de rotíferos macho B. bidentatus. Para el estudio se utilizó una cepa proveniente del bordo La Punta (22°08’ N - 102°24’ W), ubicado en Cosió, estado de Aguascalientes, México y se cultivaron en el laboratorio. Para el procesamiento de las muestras se tomaron 50 organismos fijados en Formol al 4%, de los diferentes estadios del rotífero Brachionus bidentatus, mientras que para el trofos se tomaron 25 organismos hembra del rotífero Brachionis bidentatus y se prepararon para observarse en un microscopio electrónico de barrido JEOL 5900 LV, mientras que para las observaciones de las células sexuales del macho se aislaron 500 organismos machos del rotífero Brachionus bidentatus, se fijaron e incluyeron en resina epóxica (EPON) para su observación en un microscopio electrónico de transmisión JEOL 1010 operado a 80kv. Los análisis obtenidos de hembras de B. bidentatus en cultivos de laboratorio demuestran un ciclo de vida de cinco días (4.69±0.48; N=13), y una producción de 4.5±3.67 (N=6) huevos partenogenéticos. En el caso de los huevos no fertilizados, la hembra produce más de 18 huevos (13±4.93; N=7). Para los huevos sexuales solo se produce uno solo por hembra (1±0; N=20). En el ciclo sexual, los tiempos de copula entre el macho y la hembra están en el rango de 10 a 40 segundos (17.33±10.55, N=7). Los espermatozoides se componen de un cuerpo celular y un flagelo, el tamaño del cuerpo celular es de 300nm mientras que el flagelo mide 1 700nm. Los bastones presentan una doble membrana y su tamaño a lo largo va de 2.45µm±0.74; N=6 mientras que el ancho es de 0.773µm±0.241; N=11. La evidencia sobre las características específicas ultraestructurales del rotífero B. bidentatus son notorias, más aun en el macho y en el quiste. En cuanto a la ultraestructura de los espermatozoides y los bastones respecto a otras especies sólo difieren en su tamaño, a pesar de las similitudes estructurales las estructuras de los bastones se evidencian con mayor claridad y dan más evidencias sobre su funcionalidad. Nuestro estudio de la ultraesturctura de esta especie añade información útil que junto con un análisis molecular ayudarán a clarificar la taxonomía de rotíferos brachionidos.
Assuntos
Animais , Feminino , Masculino , Óvulo/ultraestrutura , Rotíferos/ultraestrutura , Espermatozoides , México , Microscopia Eletrônica de Transmissão , Reprodução , Rotíferos/anatomia & histologiaRESUMO
BACKGROUND: Guidelines for diagnosis and therapy of meconium obstruction in extremely low birth weight neonates are still not well established. METHODS: All low-birth-weight infants presenting with meconium obstruction over a 5-year period were reviewed retrospectively. Patients with meconium plug syndrome or cystic fibrosis were excluded. RESULTS: Seven patients were identified. Average birth weight and gestational age were 874 g and 27.7 weeks, respectively. All were products of high-risk pregnancies, and 6 (86%) were delivered by cesarean section. All patients presented with distended abdomens without peritonitis. Abdominal films showed multiple distended intestinal loops without air-fluid levels in all cases. Three patients (43%) had contrast enemas, all showing microcolon. Nonoperative therapy, consisting of rectal irrigations and N-acetylcysteine per orogastric tube, succeeded in 3 patients who were obstructed for less than 10 days and failed in 1 patient obstructed for 12 days. Four patients, all obstructed for more than 10 days, underwent enterotomy and lavage (2), bowel resection and ileostomy (1), and initial peritoneal drainage for perforation followed by bowel resection (1). All patients survived with intact bowel function. CONCLUSIONS: Extremely low-birth-weight infants with meconium obstruction can be diagnosed based on their typical clinical and plain radiographic characteristics, without need for a contrast enema. Nonoperative treatment is successful early in the course of the obstruction. In the absence of an intestinal complication, simple enterotomy and meconium evacuation effectively treats long-standing obstruction.