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Biomedical subjects

C M Mendes

Publications and source records attributed to C M Mendes.

At least 19 recordsLinked to original sources

Morphological changes in mouse embryos cryopreserved by different techniques.

Cryopreservation of mammalian embryos is an important tool for the application of reproductive biotechnologies. Subjective evaluation to determine embryo viability is often used. The determination of the best cryopreservation protocol depends on morphological and molecular analysis of cellular injuries. The main objective of this study was to compare two methods of cryopreservation by assessing morphological alterations of frozen embryos using light, fluorescence, and transmission electron microscope. Fresh (control), slow frozen, and vitrified mouse embryos were composed. To evaluate the viability of the embryos, the cell membrane integrity was assessed using Hoechst33342 and propidium iodide (H/PI) staining. Morphological analyses using hematoxylin and eosin (HE) staining were performed to test different techniques (in situ, paraffin, and historesin) by both light and fluorescence microscopy. Transmission electron microscope was used to detect ultrastructural alterations in Spurr- and Araldite-embedded samples. H/PI staining detected more membrane permeability in the vitrification (69.8%) than in the slow freezing (48.4%) or control (13.8%) groups (P < 0.001). Historesin-embedded samples showed to be more suitable for morphological analyses because cellular structures were better identified. Nuclear evaluation in historesin sections showed the induction of pycnosis in slow freezing and vitrification groups. Cytoplasm evaluation revealed a condensation and an increase in eosinophilic intensity (indicating apoptosis) in the slow freezing group, and weakly eosinophilic structures and degenerated cells (indicating oncosis) in the vitrification group (P < 0.05). Ultrastructural analyses confirmed HE morphological findings. It was concluded that both cryopreservation techniques resulted in oncosis and apoptosis injuries. However, vitrification caused more severe cellular alterations and reduced embryonic viability compared to slow freezing.

Animals↗

Production of a cloned calf from a fetal fibroblast cell line.

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

Animals↗

In vitro susceptibility of gram-positive cocci isolated from skin and respiratory tract to azithromycin and twelve other antimicrobial agents.

This study was conducted to evaluate the activity of azithromycin in comparison to 12 other antibacterial agents against recent isolates obtained consecutively from patients with respiratory tract or skin infections, from January to July, 2000. A total of 717 Gram-positive cocci were analyzed in this study and the following species were studied: Staphylococcus aureus (n=576), beta-hemolytic streptococci (n=115), and Streptococcus pneumoniae (n=26). Susceptibility testing was carried out by the disk diffusion method and interpreted according to NCCLS breakpoints. The activity of azithromycin was compared to erythromycin, clindamycin, chloramphenicol, ciprofloxacin, ofloxacin, oxacillin, penicillin, ceftriaxone, tetracycline, trimethoprim/sulfamethoxazole, teicoplanin, and vancomycin. Of the 26 S. pneumoniae isolates recovered from the respiratory tract, 5 (19.2%) were intermediate resistant to penicillin. All of these strains were susceptible to chloramphenicol, ofloxacin, and vancomycin, and 24 (92%) were also susceptible to azithromycin, clindamycin, and erythromycin. Among the 67 beta-hemolytic streptococci strains isolated from the respiratory tract, 66 (99%) were susceptible to azithromycin, erythromycin, clindamycin, and ofloxacin. All 48 beta-hemolytic streptococci strains isolated from skin were susceptible to azithromycin and clindamycin, 47 (98%) were susceptible to erythromycin, and 46 (96%) were susceptible to ofloxacin. Of the 576 strains of S. aureus, 253 (43.9%) were isolated from the respiratory tract and 323 (56.1%) from skin. Among S. aureus isolates from the respiratory tract and skin, 46 (18%) and 78 (24%), respectively were resistant to oxacillin. Isolates from the respiratory tract and skin showed the same percentage of resistance (36%) to azithromycin. These in vitro results suggest that azithromycin can be a therapeutic option for treatment of infections caused by these bacteria since the newer macrolides have several distinct advantages over erytromycin including improved oral bioavailability, longer half-life allowing once or twice daily administration, higher tissue concentrations and less gastrointestinal adverse effects.

Anti-Bacterial Agents↗

In vitro activity of fluoroquinolones (gatifloxacin, levofloxacin and trovafloxacin) and seven other antibiotics against Streptococcus pneumoniae.

In recent years, the level of resistance of S. pneumoniae to beta-lactam and/or macrolides has increased around the world including some countries in South America. Because of this resistance, it is necessary to test the therapeutic alternatives for treating this pathogen, including the newer quinolones. This study was carried out in order to compare the in vitro activity of fluoroquinolones gatifloxacin, levofloxacin and trovafloxacin, to penicillin G, amoxicillin, amoxicillin-clavulanate, cufuroxime sodium, ceftriaxone, azithromycin and clarithromycin, against 300 strains of S. pneumoniae. Of the 300 samples tested, 18.6% were not susceptible to penicillin (56 strains) and 7% (21 strains) were resistant to the second generation cephalosporin. Among the macrolides, resistance ranged from 6.7% for clarithromycin to 29.6% for azithromycin. Susceptibility to the newer quinolones was 100% including the 56 strains not susceptible to penicillin. Among the 10 antibiotics evaluated, the fluoroquinolones gatifloxacin, levofloxacin, and trovafloxacin displayed high levels of in vitro activity against S. pneumoniae.

Anti-Bacterial Agents↗

Left ventricular mass and function in young obese women.

OBJECTIVE: Obesity is associated with a high mortality rate due to cardiovascular disease. Left ventricular (LV) hypertrophy has been described in relation to obesity. The aim of this study was to evaluate echocardiographically the LV mass and function in young obese women as compared to lean women with similar characteristics. DESIGN: Prospective study. SUBJECTS: Eighty-two young women (< or =40 y), with obesity degree varying from I to III (BMI from 30 to 50 kg/m2) were compared to eighty young lean women. All of them were normotensive, none had cardiovascular complaints or any previous history of pulmonary disease, and none were taking any medication. The LV mass was calculated by the Devereux and Reichek formula. RESULTS: The LV mass was strongly increased in all obese groups (P<0.00003 to 0.000005) compared to lean subjects. LV mass adjusted indexes for height, BMI or volume were also increased compared to lean subjects and when adjusted for weight it was decreased. However when comparing LV mass/body surface area index this difference was not statistically significant. The linear regression analysis showed a strong association between the degree of obesity and LV mass, (r=0.52, P<0.001). Systolic and diastolic function in obese patients were similar to lean subjects, except for a lower E/A ratio in the obese group (P=0.005). CONCLUSION: In asymptomatic young obese women, there are some echocardiographic findings suggesting early cardiac involvement that seems to be related to the degree of obesity.

Adult↗

Environmental contamination by multidrug-resistant Acinetobacter baumannii in an intensive care unit.

Contamination in an intensive care unit caused by multidrug-resistant Acinetobacter baumannii complex (MRAB)-colonized patients was evaluated using environmental and patient cultures. MRAB occurred in 21% of patients' cultures, 2.1% of 513 areas surrounding MRAB-patients, and one of 372 common areas. No transmission to other patients occurred. Barrier precautions and ethanol disinfection may prevent dissemination.

Acinetobacter Infections↗

Nosocomial infections caused by multiresistant Pseudomonas aeruginosa.

A case-control study was done to evaluate factors associated with nosocomial infections by multiresistant Pseudomonas aeruginosa (MRPA). Results showed that MRPA was associated with the use of immunosuppressive and antimicrobial drugs. Five typing methods indicated that the MRPA infections were due to multiple strains rather than a single strain.

APACHE↗

Evaluation of the in vitro activity of cefepime compared to other broad-spectrum cephalosporins against clinical isolates from eighteen Brazilian hospitals by using the Etest.

The in vitro activity of cefepime was compared to that of ceftazidime, ceftriaxone, and cefotaxime in a multicenter study involving 10 clinical microbiology laboratories and clinical isolates from 18 Brazilian hospitals from 7 cities (4 states). A total of 982 isolates consecutively collected between December 1995 and March 1996 were susceptibility tested by using Etest and following the NCCLS procedures for agar diffusion tests. The cefepime spectrum was broader than that of the other broad-spectrum cephalosporins against both Gram-negative rods and Gram-positive cocci. Cefepime was particularly more active against Enterobacter sp. (MIC90, 2 micrograms/ml), Serratia sp. (MIC90, 2 micrograms/ml) and oxacillin-susceptible Staphylococcus aureus (MIC90, 3 micrograms/ml). Against Pseudomonas aeruginosa, cefepime (MIC90, 16 micrograms/ml) was slightly more active than ceftazidime (MIC90, 32 micrograms/ml) and 8- to 16-fold more active than ceftriaxone of cefotaxime (MIC90, > 256 micrograms/ml). Our results show that nosocomial bacteria, especially Gram-negative rods, have a high rate of cephalosporin resistance in Brazil. However, part of these resistant bacteria remains susceptible to cefepime. The Etest was shown to be an excellent method for multicenter studies of the in vitro evaluation of new antimicrobial agents.

Bacteria↗

In vitro activity of ampicillin-sulbactam against clinical multiresistant Acinetobacter baumannii isolates.

We evaluated the in vitro activity of ampicillin-sulbactam in comparison with that of broad-spectrum antimicrobial agents against Acinetobacter baumannii isolates. Two hundred and twelve clinical isolates collected between January 1993 and March 1995 from two tertiary hospitals located in São Paulo, Brazil were tested for susceptibility by the disk diffusion method against several broad-spectrum antimicrobial agents, including imipenem, ciprofloxacin, ceftazidime, aztreonam, amikacin, and polymyxin B. All strains were susceptible to polymyxin B. The second most active compound was the combination ampicillin-sulbactam (88% susceptibility). Only 79% of the isolates were susceptible to imipenem. Ciprofloxacin was active against 60 (28%) and amikacin against 34 (16%) isolates. Ceftazidime was the most active cephalosporin; however, only 9% of the isolates were susceptible to this compound. Both aztreonam and ampicillin alone showed very poor activity against this species (1% susceptibility). The prevalence of severe infections due to A. baumannii is increasing very rapidly in the tertiary hospitals of São Paulo and there are very few options for the treatment of these infections. Polymyxin B is invariably in vitro active against this species; however, this compound can cause severe side effects and is not commercially available for intravenous use in Brazil and in several other countries. Our results indicated that the combination ampicillin-sulbactam may be an alternative drug for the treatment of infections due to multiresistant A. baumannii; however, further studies are necessary to evaluate the clinical role of this compound for the treatment of severe infections.

Acinetobacter↗

Electric showers as a control measure for Legionella spp. in a renal transplant unit in São Paulo, Brazil. Legionellosis Study Team.

After an outbreak of legionnaires' disease Legionella pneumophila serogroup 1 in a renal transplant unit in São Paulo, Brazil, periodic hyperchlorination and flushing of pipes were instituted as control measures. These were only partially effective as every two to five months water cultures turned positive or new cases of the disease occurred. In November 1993 the hot water was disconnected from the unit and small, plastic electric showers were installed in each bathroom. Over a period of 12 months water from showers and taps was cultured for Legionella spp. every two weeks. On only one occasion was a water culture positive for L. pneumophila from a sink tap. No water sample obtained from showers was positive during the study period. No cases of legionnaires' disease occurred. We considered the use of electric showers an inexpensive and effective method of controlling the problem of Legionella spp. in the water system of our renal transplant unit.

Baths↗

Epidemiologic investigation of an outbreak of coagulase-negative Staphylococcus primary bacteremia in a newborn intensive care unit.

Infections due to coagulase-negative Staphylococcus (CNS) are an ever-increasing nosocomial problem, particularly in the pediatric population. The authors describe a cluster of three primary bloodstream infections due to CNS in a newborn intensive care unit that occurred between November 23 and December 2, 1992. Two children died as a direct consequence of the bacteremia; at autopsy, one had a large bacteria-containing thrombus extending from the insertion site of a central catheter to the superior vena cava. The children were placed in isolation, and the nursing and medical staff were given topical nasal mupirocin. Plasmid analysis performed later disclosed three different blood isolates that also were different from any of the staff's nasal isolates. The authors concluded that molecular methods such as plasmid analysis are important tools in identifying true outbreaks and can prevent needless interventions, such as those during this cluster.

Bacteremia↗

An outbreak of nosocomial Legionnaires' disease in a renal transplant unit in São Paulo, Brazil. Legionellosis Study Team.

From June 1989 to March 1990 there were eight cases of Legionnaires' disease caused by Legionella pneumophila serogroup 1 in a renal transplant unit. There were seven cases of pneumonia and one case of pleural effusion. A study was conducted to identify the source of the outbreak. Legionella anisa was cultured from tap water. Twenty-seven staff members of the unit were serologically tested and antibody titres were positive in two. The probable source of infection was the potable water system. Control measures were hyperchlorination and heating of the water, after which there were no further cases during 5 months' follow up. We believe this is the first reported Legionnaires' disease outbreak in Latin America.

Brazil↗

Campylobacter fetus endocarditis on a prosthetic heart valve.

Campylobacter fetus infection in man is rare. Few cases of Campylobacter endocarditis have been described, and none have been associated with a prosthetic heart valve. We have reported a case of Campylobacter fetus ssp fetus on a prosthetic mitral valve in a 48-year-old woman who was treated successfully with penicillin and streptomycin (and later with gentamicin). Replacement of the prosthetic valve was done because of stenosis and vegetations. Pathologic examination showed vegetations on a calcified prosthesis, but no bacteria were seen.

Campylobacter Infections↗

Rapid, automated identification of novobiocin-resistant, coagulase-negative staphylococci.

A modified automated method that uses the MS-2 system (Abbott Laboratories, Diagnostics Div., Irving, Tex.) to verify the reaction of coagulase-negative staphylococci to novobiocin is described. This technique permits the testing of a great number of specimens in an average time of 99 min and results in a 100% match with the traditional method of culturing.

Bacteriological Techniques↗