Histomorphometry of preterm and term human placentas / Histomorfometría de placentas de pretérmino y término

The intra-uterine existence of foetus is dependent on placenta, a major organ of nutrition and homeostasis.The present study was carried out to compare morphometric and histological changes in preterm and term human placentas. Eighty placentas collected from Department of Obstetrics and Gynecology, JNMCH, AMU, Aligarh, were divided into group first of preterm placentas up to 36 weeks (n =30) and second group of full term placentas i.e. 37 to 40 weeks ( n = 50) respectively. The samples were fixed in 10 percent formol-saline solution. The gross morphological variables of placentas were studied. There was a significant increase in the placental weight, decidual area and umbilical cord diameter of term placenta as compared to that of the preterm ones. From each placenta whole thickness tissue blocks were taken and processed for paraffin sectioning. Five µ-thick sections were stained with Haematoxylin-eosin and Van Gieson stains and processed for light microscopy. A total of 200 villi were studied in each sample under high power field and occurrence of different features was expressed as percentages for each parameter. The appearance of microvilli and syncytial bud on the syncytium were almost absent in the villi of term placentas. It was concluded that with increasing gestational age there was a gradual increase in the number of capillaries in villi from preterm to term placenta.There was a significant increase in syncytial knot count, fibrinoid necrosis, vasculosyncytial membrane and decrease in the percentage of villi showing cytotrophoblastic cells and number of Hofbauer cells in term group as compared to preterm group.

La existencia intrauterina del feto depende de la placenta, el mayor órgano de nutrición y homeostasis. El estudio se llevó a cabo para comparar los cambios morfométricos e histológicos de la placenta humana de término y pretérmino. Ochenta placentas fueron obtenidas del Departamento de Obstetricia y Ginecología, JNMCH, AMU, Aligarh y se dividieron en grupos, el primer grupo de placentas de pretérmino hasta 36 semanas (n = 30) y el segundo grupo de placentas de término, de 37 a 40 semanas (n = 50 ). Las muestras fueron fijadas en solución de formol-salina al 10 por ciento. Se estudiaron las variables morfológicas macroscópicas de las placentas. Hubo un aumento significativo en el peso de la placenta, el área de decidua y el diámetro del cordón umbilical de la placenta a término en comparación con la de los prematuros. De cada placenta se tomaron y se procesaron bloques de tejido para incluirlos en parafina. Cortes de 5 µm fueron teñidos con HE y Van Gieson para microscopía óptica. De cada muestra fueron estudiadas 200 vellosidades, bajo campo de alta resolución y la aparición de diferentes características se expresó como porcentajes para cada parámetro. La aparición de las microvellosidades y brote sincitial en el sincitio estaban casi ausente en las vellosidades de las placentas de término. Se puede concluir que al aumentar la edad gestacional hubo un aumento gradual en el número de capilares en las vellosidades de la placenta de término. Existe un aumento significativo en el recuento de nudo sincitial, necrosis fibrinoide, membrana vasculosincisial y disminución en el porcentaje de las vellosidades que muestran células citotrofoblástica y número de células de Hofbauer en las placentas del término de grupo en comparación con el grupo de pretérmino.

Transformation of coffee (Coffea Arabica L. cv. Catimor) with the cry1ac gene by biolistic, without the use of markers.

The transformation of coffee plantlets with the cry1ac gene of Bacillus thuringiensis was achieved by biolistic using either the whole pUBC plasmid or only the ubi-cry1ac-nos genetic cassette. The cry1ac gene was inserted into coffee plants in order to confer resistance to the leaf miner Leucoptera coffeella, an insect responsible for considerable losses in coffee crops. Bearing in mind that the genetic cassettes used for this study lack reporter genes and/or selection marker genes, the parameters for the transformation procedure by biolistic were previously standardised with a plasmid carrying the gus reporter gene. The presence of the cry1ac gene in young plantlet tissues was determined by PCR, Southern blot and reverse transcription-PCR. Our results show that the obtainment of viable coffee plantlets, transformed by bombardment with the cry1ac gene and without selection markers nor reporter genes, is feasible.

Transformation of coffee (Coffea Arabica L. cv. Catimor) with the cry1ac gene by biolistic, without the use of markers

The transformation of coffee plantlets with the cry1ac gene of Bacillus thuringiensis was achieved by biolistic using either the whole pUBC plasmid or only the ubi-cry1ac-nos genetic cassette. The cry1ac gene was inserted into coffee plants in order to confer resistance to the leaf miner Leucoptera coffeella, an insect responsible for considerable losses in coffee crops. Bearing in mind that the genetic cassettes used for this study lack reporter genes and/or selection marker genes, the parameters for the transformation procedure by biolistic were previously standardised with a plasmid carrying the gus reporter gene. The presence of the cry1ac gene in young plantlet tissues was determined by PCR, Southern blot and reverse transcription-PCR. Our results show that the obtainment of viable coffee plantlets, transformed by bombardment with the cry1ac gene and without selection markers nor reporter genes, is feasible.

A transformação das plântulas de café com o gene cry1ac de Bacillus thuringiensis foi realizada por biobalística, utilizando todo o pUBC plasmídeo ou só o cassete genético UBI-cry1ac-nos. O gene cry1ac foi inserido no cafeeiro a fim de conferir resistência à folha mineiro Leucoptera coffeella, um inseto responsável por perdas consideráveis nas culturas de café. Tendo em conta que ao plasmídeo e ao cassete genético utilizados para este estudo faltam genes repórteres e/ou de seleção, os parâmetros para o processo de transformação por biobalística foram previamente padronizados com um plasmídeo transportando o gene repórter gus. A presença do gene cry1ac em tecidos de jovens plântulas foi determinada por PCR, Southern blot e transcrição reversa-PCR. Nossos resultados mostram que a obtenção de plântulas de café, transformado por bombardeamento com o gene cry1ac sem genes de seleção genética nem repórteres é viável.

[Epidemic of pneumonia associated with mechanical ventilation in Mérida, Yucatán]. / Epidemia de neumonía asociada a ventilación mecánica en Mérida, Yucatán.

OBJECTIVE: To determine the main epidemiological, clinical, and microbiologic characteristics of an outbreak of ventilator-associated pneumonia at an intensive care unit in Yucatan. MATERIAL AND METHODS: An 11-month prospective and observational study was designed to determine incidence, mortality, potential reservoirs, etiologic agents and antibiotic susceptibility patterns. RESULTS: The incidence of ventilator-associated pneumonia was 74%. The crude mortality rate was 88% compared to a 19.5% expected-mortality rate. Gram-negative bacteria were isolated from 98% of the cultures, of which 46% were susceptible to third generation cephalosporins, 59% to fourth generation cephalosporins, 70% to ciprofloxacin and 100% to imipenem. Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated from some of the ventilator circuits and the sink. CONCLUSIONS: The high incidence of pneumonia and associated mortality in our intensive care unit may be attributed to the absence of infection control measures and the high prevalence of multiresistant organisms which is related to antibiotic abuse.

Disinfection and sterilization practices in Mexico.

We evaluated antisepsis, disinfection, and sterilization procedures at 22 hospitals in the state of Yucatan, Mexico, which provide services for approximately 80% of the population. The percentage of hospitals that followed standard recommendations for diverse antisepsis procedures were as follows: surgical scrub, 41%; surgical site preparation, 68%; central intravenous (iv) catheters, 61%; peripheral iv catheters, 86%; urinary catheters, 41%, and umbilical cord care, 5%. Inappropriate procedures typically involved the use of benzalkonium chloride and mercury compounds. Adequate sterilization procedures were observed for sharp surgical instruments in 9%, for blunt surgical instruments in 81%; for linen in 90%; for surgical brushes in 9%; for metal prostheses in 100%; for plastic prostheses in 57%; and for disinfection of endoscopes in 30%. The most common errors were the use of too short an exposure time in steam sterilizers or dry heat sterilizers, and the use of benzalkonium chloride as a sterilizing agent. Only 14% of hospitals used commercial spore preparations for autoclave monitoring. For the reuse of disposables, 50-94% of hospitals employed inappropriate sterilization or low-level disinfection procedures. Similar conditions are likely to exist in hospitals throughout the country. The establishment of strict regulations and hospital training programmes for disinfection and sterilization procedures, and the reuse of disposable devices is urgently needed in Mexico.

[7 years' experience with neonatal tetanus in Yucatan]. / Experiencia de siete años con tétanos neonatal en Yucatán.

Forty-six patients with neonatal tetanus (NT) were studied to identify factors for poor prognosis which could be used to select a therapeutic regimen with intravenous diazepam or neuromuscular blockade (NB) with pancuronium. Nine patients with NT grades II and III were successfully treated with diazepam; none of them died. Of the patients with NT grades IV and V, 27 received diazepam and ten received pancuronium. The mortality rate in these groups was 55% and 100%, respectively. Factors predicting poor prognosis among patients treated with diazepam were apneas (P = 0.01), and an age on admission of seven days or less (P = 0.0002). Patients who received diazepam and died, generally presented a rapidly fatal course (mean of four days); in this group tetanus was the main cause of death (73%). Patients treated with pancuronium survived a comparably longer period (mean = 15.7 days, P = 0.05), but generally died from nosocomial infections (70%, P = 0.04). On the basis of our results we propose that NT grades IV and V with the aforementioned factors for poor prognosis be treated with NB. In hospitals with limited resources and high rates of nosocomial infection, we suggest that NT grades IV and V without such factors initially be managed with diazepam, reserving NB for therapeutic failures. Finally, NT grades I-III may be effectively treated with diazepam alone.

Hospital-acquired diarrhea in adults: a prospective case-controlled study in Mexico.

OBJECTIVE: To know the incidence, etiology, risk factors, morbidity, and mortality of nosocomial diarrhea in adults. DESIGN: Nested case-control study, matched by service, length of stay, date of admission, and presence of leukopenia and/or the acquired immunodeficiency syndrome (AIDS). Cases were those who developed nosocomial diarrhea. Controls were those who did not develop nosocomial diarrhea during a comparative period nor during the next ten days. Stool samples were processed in search for parasites, yeasts, bacteria, and rotavirus. SETTING: Third-level referral center, in Mexico City, Mexico, for general internal medicine and surgical problems. PATIENTS: Eligible subjects were all new admissions to the hospital from November 1987 to September 1988. Reasons for exclusion were presence of chronic diarrheal disease or melena. There were 115 cases and 111 controls. RESULTS: Overall risk of acquiring nosocomial diarrhea was 5.5%, or 1.8 episodes per 100 patient-weeks. A potential pathogen was found in 59%. Yeasts and Entamoeba histolytica were the most frequently isolated pathogens. Mortality in cases was 18%, as compared with 5% in controls (p less than .01). Multivariate analysis showed enteral feeding, recent enemas, presence of Candida species, use of antacids/H2-blockers, and presence of nasogastric tubes as significant risk factors for nosocomial diarrhea. CONCLUSIONS: Diarrhea is a common complication in hospitalized patients. It occurs more often than previously suspected and is linked with a substantial mortality. The spectrum of etiologic agents is different from that reported in pediatric hospitals. Given that nosocomial diarrhea may constitute, at least, a marker of severity of illness, it should receive more attention in general hospitals.

Epidemic of Serratia marcescens bacteremia and meningitis in a neonatal unit in Mexico City.

A case-control study was conducted on an epidemic of bacteremia and meningitis caused by Serratia marcescens in the neonatal intensive care unit and special care nursery of a general hospital in Mexico City, Mexico. A 19.9% incidence of bacteremia and meningitis was recorded in contrast to 1.4% and 3.7% during preepidemic and post-epidemic periods; a 69% mortality rate was observed. Peripheral IV catheters and the use of mixed IV fluids prepared in the wards were the major risk factors (P less than 0.001). Rectal and nasopharyngeal cultures were positive in 68% of asymptomatic neonates and hand cultures were positive in 16.7% of personnel. Strains were resistant to all aminoglycosides and broad-spectrum penicillins, and belonged to the A5/8 biogroup. Containment of this outbreak was difficult because of failure to identify colonized infants early in the epidemic and because of persistent carriage of S marcescens by personnel. Comparisons between this hospital and tertiary care centers in Mexico suggest that in developing countries nosocomial infections could be of greater magnitude in secondary than in tertiary level centers.