Efectividad de la profilaxis antibiótica en pacientes con colecistitis aguda sometidos a colecistectomía laparoscópica / Effectiveness of antibiotic prophylaxis in acute cholecystitis patients undergoing laparoscopic cholecystectomy

OBJETIVO: Determinar la efectividad de la profilaxis antibiótica con Cefazolina en pacientes sometidos a colecistectomía laparoscópica por Colecistitis Aguda MATERIAL Y MÉTODOS: Cohorte Prospectiva POBLACIÓN: Pacientes mayores de 18 años, con patología litiásica vesicular aguda, sometidos a colecistectomía laparoscópica. SEDE Y TEMPORALIDAD: Hospital Obrero Nº 1 de la Caja Nacional de Salud La Paz ­ Bolivia. Período comprendido entre el 1 de Julio de 2016 al el 31 de Diciembre de 2016. RESULTADOS: Se incluyeron 95 pacientes con Colecistitis Aguda divididos en dos grupos, el Grupo A (SIN profilaxis antibiótica) compuesto por 50 sujetos y el Grupo B (CON profilaxis antibiótica) de 45 sujetos. La edad promedio fue de 48 años, el peso de 70 kilos, la talla de 165 cm y el IMC de 27,4 km/ m2. El tiempo operatorio promedio fue de 50 (±22,815) minutos en el total del grupo, 45 min. (±18,460) en el grupo A y 60 min (±24,862) en el grupo B. La conversión a cirugía abierta fue de 9 sujetos (9,5%). La infección del sitio operatorio se presentó en 47 sujetos (49,5%), 30 sujetos (60%) EN EL GRUPO A y 18 en el grupo B (40%). El OR calculado es de 0,444 (IC 95% 0,195 ­ 1,011). CONCLUSIONES: La administración de Cefazolina en forma profiláctica, parece no disminuir la probabilidad de infección del sitio operatorio en colecistitis aguda abordada por laparoscopía

OBJECTIVE: To determine the effectiveness of antibiotic prophylaxis with Cefazolin in patients undergoing laparoscopic cholecystectomy due to Acute Cholecystitis. METHODS: Prospective Cohort POPULATION: Adult patients (older than 18 years), with acute lithiasic cholecystitis, who underwent laparoscopic cholecystectomy. PLACE AND TEMPORALITY: Hospital Obrero No. 1 of the Caja Nacional de Salud La Paz ­ Bolivia, from July to December 2016. RESULTS: A total of 95 patients with Acute Cholecystitis were enrolled and divided in to two groups, group A (without antibiotic prophylaxis) composed of 50 subjects and Group B, (with antibiotic prophylaxis) 45 subjects. The mean age was 48 years old, weight 70 Kg, hight 165 cm and a BMI of 27.4 kg/M2. The mean operating time was 50 minutes (+- 22.185), group A 45 minutes and group B 60 min. Conversion to open surgery happened in 9 patients (9,5%), all in group B. Surgical Site infection (SSI) occurred in 47 patients (49.5%), of whom 30 patients belong to group A (60%) and 18 patients to group B (40%). The calculated Odds ratio is 0.444 (IC 95% 0,195-1.011). There were no bile duct injuries or morality in this study. CONCLUSIONS: The prophylactic administration of Cefazolin does not seems to decrease the probability of SSI in acute cholecystitis treated laparoscopically.

Satoyoshi syndrome with unusual skeletal abnormalities and parental consanguinity.

Satoyoshi syndrome (SS) (OMIM 600705) is a rare multisystemic disorder of unknown etiology characterized by progressive painful intermittent muscle spasm, alopecia universalis, diarrhea, short stature, amenorrhea, and secondary skeletal abnormalities mimicking a metaphyseal chondrodysplasia. To date all reported cases have been sporadic. We describe a 26-year-old Mexican woman, a product of consanguineous parents with clinical characteristics of SS. Our patient, also showed skeletal anomalies not previously reported that seems to be a coincidental finding.

Advances in the development of molecular tools for the control of bovine babesiosis in Mexico.

The severe negative impact that bovine babesiosis has in the Mexican cattle industry has not been ameliorated basically due to the lack of safe and effective commercially available vaccines and sensitive and reliable diagnostic tests. In recent years, the Bovine Babesiosis Laboratory at the National Center for Disciplinary Research in Veterinary Parasitology-INIFAP in Morelos State, Mexico has been directing efforts towards three main research areas: (1) The development of in vitro culture-derived, improved and safer live vaccines. This has been done in two ways: using gamma-irradiated bovine serum and erythrocytes for the in vitro culture of vaccine strains, which reduces the risk of contaminating pathogens, and improving the immune response, by the addition of L. casei, a strong stimulant of the innate immune system. (2) The study of antigens considered as vaccine candidates with the goal of developing a recombinant vaccine that suits the country's needs. Knowing their degree of conservation or variation in Mexican isolates, their phylogenetic relationship and their protective, immuno-stimulatory properties, are first steps towards that goal. (3) The development of new tools for diagnosis, detection and discrimination of bovine babesiosis is the third area. Developing variants of ELISA, which are more reliable than the currently used IFAT, are a priority, and finally, taking advantage of the genomes of Babesia bigemina, and B. bovis, we are identifying genes than allow us to discriminate isolates using molecular tools.

Trisomy 3q25.1-qter and monosomy 8p23.1-pter in a patient: cytogenetic and molecular analysis with delineation of the phenotype.

We describe a 4-year-old boy with partial 3q trisomy and distal 8p monosomy. The patient presented with mental retardation, dysmorphic face, congenital heart defect, brain and genital anomalies, and behavioral problems. The conventional cytogenetic analysis showed a 46,XY,add(8p) karyotype. Reverse painting and microsatellite analysis demonstrated a partial monosomy of 8p23.1 --> pter and a partial trisomy of 3q25.1 --> qter. The data suggest that the chromosomal rearrangement originated from a de novo translocation in a paternal germinal cell. The phenotype observed in our patient resulted from the combination of those defects described in the isolated dup(3q) and distal del(8p) syndromes.

Bovine babesiosis and anaplasmosis follow-up on cattle relocated in an endemic area for hemoparasitic diseases.

A multiplex PCR/DNA probe assay was used to monitor Babesia bovis, B. bigemina and Anaplasma marginale infection in cattle introduced to a Boophilus microplus-infested area in Veracruz, Mexico. Eight intact, 18-month-old, cross-bred beef cattle (four naive, Group A; four Babesia species--premunized, Group B) were immediately exposed to ticks after arrival and were clinically monitored from day 6 to day 98 post-exposure (PE) to ticks. Blood sample analysis for DNA detection by the MPCR/DNA probe assay showed that Group A animals were infected with B. bovis from day 11 up to day 22 PE, requiring treatment on days 17-20. Group B animals were detected positive to B. bovis on days 17-20, did not require treatment and remained persistently infected from days 70 to 84 PE. Treatment of Group A animals delayed the infection with B. bigemina. These animals became positive to the parasite on days 63-77 PE. In contrast, Group B animals (untreated) showed B. bigemina infection on days 21-26 and 63-84 PE. One animal was positive for A. marginale infection on days 63-66 PE, the rest of the animals became so on days 80-98 PE. All infected animals required treatment with oxytetracycline. Monitoring the triple hemoparasite infection with the MPCR/DNA probe assay provided important epidemiological information. Thus, precautionary measures can be established when cattle are moved to a babesiosis/anaplasmosis risk area.

Use of a duplex PCR/DNA probe assay to monitor Babesia bovis and Babesia bigemina in cattle during a vaccination trial.

A Duplex Polymerase Chain Reaction (DPCR)/DNA probe assay was used to detect Babesia bovis and B. bigemina DNA in cattle undergoing immunization trials. Blood samples were collected from 15 non-splenectomized, 1-2 years old bulls, inoculated with 1 x 10(7) each of culture-derived B. bovis- and B. bigemina-infected erythrocytes. 15 bulls inoculated with normal erythrocytes served as a control group. All cattle were field exposed to tick-transmitted Babesia 21 days (20 animals, Group I) and 60 days (10 animals, Group II) post-inoculation (PI). After immunization, the DPCR/DNA probe assay detected B. bigemina and B. bovis parasite DNA in all inoculated animals from days 4 to 14 PI. At challenge, B. bovis DNA was detected in all control animals as early as day 8 (Group I), or day 11 (Group II) post-introduction to a tick-infested area. The immunized bulls showed B. bovis positive PCR/DNA probe signals from day 0 (Group II) and day 8 (group I), up to day 32 post-exposure to ticks. Positive B. bigemina signals were detected from day 0 (Group I) and day 8 (Group II), up to day 36 post-exposure to ticks. During challenge, it was not possible to clearly define whether the PCR/DNA probe signals detected in the blood from immunized cattle were a result of amplified DNA from the culture-derived parasites, from the tick-transmitted parasites, or both.

Molecular epidemiology of bovine anaplasmosis.

Bovine anaplasmosis presents a worldwide distribution. However, specific models for studying the epidemiology of the disease are not available. Epidemiological modeling encounters some difficulties due to a lack of culturing techniques for Anaplasma marginale, the causative agent, as well as for the lack of typing techniques to characterize strains. The chronic carrier state and the population dynamics of mechanical and biological vectors also create difficulties. In addition, conventional serology and blood smear diagnostic techniques fail to detect all chronic carriers. Fortunately the needs for the accurate typing of isolates and for detecting chronic carriers made it possible to encourage the development of new tools based on molecular epidemiology principles. A. marginale isolates can now be typed by using panels of monoclonal antibodies, and the genes coding for some major surface proteins can be expressed or analyzed by looking at the nucleotide arrangement level. In the same manner, the latest techniques for detecting A. marginale chronic infections use DNA and RNA probes, and PCR-based methods to detect A. marginale DNA from bovine blood samples with extremely low rickettsaemias. Currently all these new epidemiological tools are being incorporated to experimental models to analyze their applicability for epidemiological studies in the near future.

Use of multiplex polymerase chain reaction-based assay to conduct epidemiological studies on bovine hemoparasites in Mexico.

A study was conducted to test the applicability of a Polymerase Chain Reaction (PCR)-based approach for the simultaneous detection of the bovine hemoparasites Babesia bigemina, B. bovis and Anaplasma marginale. Bovine blood samples from cattle ranches of a previously determined enzootic zone in the Yucatan Peninsula of Mexico, were collected from peripheral blood and processed for PCR analysis. Blood samples were subjected to DNA amplification by placing an aliquot in a reaction tube containing oligonucleotide primers specific for DNA of each hemoparasite species. The PCR products were detected by Dot-Blot nucleic acid hybridization utilizing nonradioactive, species-specific, digoxigenin PCR-labeled DNA probes. Four hundred twenty one field samples analyzed by the multiplex PCR-DNA probe assay showed 66.7%, 60.1% and 59.6% prevalence rates for B. bigemina, B. bovis and A. marginale, respectively. The multiplex PCR analysis showed that animals with single, double or triple infection could be detected with the parasite specific DNA probes. The procedure is proposed as a valuable tool for the epidemiological analysis in regions where the hemoparasite species are concurrently infecting cattle.

Antibody response to Babesia bigemina infection in calves measured by ELISA and immunoblotting techniques.

To measure the antibody response to Babesia bigemina with an ELISA test, three groups of cattle were experimentally infected with two isolates of the parasite. It was possible to demonstrate specific antibody binding directed against the parasite as early as the 7 days postinfection (PI). The highest level of antibody was obtained around day 1 to 23 and remained detectable for 260 days. Challenge of the animals 260 days PI with a tick-induced B. bigemina infection depicted that homologous strain-challenged calves did not show an increase of IgG antibody levels, where as those challenged with the heterologous isolate did. In the latter groups the resulting level of antibodies was even higher than after the primary infection. The immunoblotting technique showed that the antibody response is probably directed against groups of B. bigemina components with a relative mobilities of 68-64 kDa, 62-54 kDa and 52-42 kDa, which appear to be major components of the protozoa. By observing the cross-reacting antigenicity among seven B. bigemina isolates, it was demonstrated that these components are not isolate-restricted.

Evaluation of a colorimetric Babesia bigemina-DNA probe within an epidemiological survey.

An epidemiological survey was conducted in southeast Mexico, in an effort to establish the serological reactivity and carrier status to Babesia bigemina of an indigenous cattle population. The prevalence was obtained through the Indirect Fluorescent Antibody Test (IFAT), using an in vitro culture-derived B. bigemina antigen. A specific, digoxigenin-coupled, approximately 6 Kb B. bigemina-DNA probe (BBDP), was used to indicate the presence of the parasite. Serum samples from 925 animals of all ages, were obtained within the three regions (I, II, III) of the state of Yucatan and tested by IFAT. In addition, whole blood samples drawn from 136 of the same animals of region II were analyzed using the BBDP. Positive IFAT (IFAT+) reactions were observed in 531 sera for a 57% overall prevalence. Regional values were: I = 157+ (56%), II = 266+ (68%) and III 108+ (42%). Only 32 (23%) of the blood samples tested with BBDP showed distinctive hybridization signal, in contrast with 100 (73%) IFAT+ animals. The response distribution for IFAT vs. BBDP was: +/+ 23, +/- 77, -/+ 9 and -/- 27 respectively. It was found that the analytical sensitivity of BBDP appears to be low for its utilization in widespread epidemiological surveys. It was considered, however, that the colorimetric probe might be useful to safely detect transmission prone carriers, since it is able to detect parasitemias as low as 0.001%.

Cloning of in vitro propagated Babesia bigemina.

The original Babesia bigemina culture conditions were modified with regard to infected bovine erythrocyte concentration and atmospheric environment. A procedure was designed which would yield a homogeneous parasite population, beginning with a single infected erythrocyte. Calculated dilutions were made in 96-well tissue culture plates to approach one infected erythrocyte per four wells. Growth of parasites in wells was detected between 16 and 28 days after cultures were initiated. Clones were transferred to 24-well tissue culture plates for regular maintenance. Three primary clones were selected for additional recloning. The probability that the parasites detected in one well are the progeny of a single infected erythrocyte approaches 0.99 for tertiary clones.

Enzymatic characterization of Babesia bovis.

Agarose gel electrophoresis was used to identify metabolic enzymes in Babesia bovis and B. bigemina. Glutamate dehydrogenase, lactate dehydrogenase, glucose phosphate isomerase, and hexokinase were identified in B. bovis- and B. bigemina-infected erythrocytes and B. bovis merozoite preparations. A specific electrophoretic mobility was observed for each enzyme. Malate dehydrogenase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and adenylate kinase were only detected in normal erythrocyte preparations. Inter-species, but not intra-species, variation was noted when comparing electrophoretograms of both species. Kinin-activating activity was not detected in B. bovis-infected erythrocyte or merozoite preparations at pH 4.2 or 7.6.

Concentration and enzyme content of in vitro-cultured Babesia bigemina-infected erythrocytes.

Clones of in vitro-cultured Babesia bigemina-infected erythrocytes were concentrated by several density gradient procedures. The density range of infected erythrocytes containing pairs of parasites was 1.077 to 1.089 g/ml, whereas the density range of infected erythrocytes containing single parasites was 1.092 to 1.100 g/ml. Three enzymes--lactate dehydrogenase, glucose-phosphate isomerase, and glutamate dehydrogenase--were found associated with infected erythrocytes. The parasite-specific enzyme and/or isoenzymes were shown to have different mobility patterns in starch gel electrophoresis from those found in the normal bovine erythrocytes. The enzyme 6-phosphogluconate dehydrogenase was not detected as a parasite-specific enzyme in B. bigemina-infected erythrocytes.

Babesia bovis: purification and concentration of merozoites and infected bovine erythrocytes.

Babesia bovis merozoites, externalized by removal of infected erythrocytes from ordinary culture conditions, were completely separated from red blood cells and stroma by centrifugation in a Percoll gradient. A merozoite band formed at a point corresponding to about 1.087 g/ml specific density. Infected red blood cells were concentrated approximately fourfold to obtain greater than 49.0% parasitemia after centrifugation in Percoll. Most highly enriched fractions positioned between 1.121 and 1.123 g/ml specific density. Full parasite viability was retained.

In vitro cultivation of Babesia bigemina.

A strain of Babesia bigemina was isolated from an infected calf and propagated in vitro. Culture conditions included washing of infected and normal bovine erythrocytes in a special solution, and the use of a 5% to 10% (v/v) erythrocyte suspension in medium 199 (with 20% to 50% fresh normal bovine serum) at a depth of 4 mm in a 5% CO2, 2% O2, 93% N2 atmosphere. After 36 days in vitro and 9 subcultures, the cultured organism was inoculated into a susceptible calf. This calf developed clinical signs of disease and recovered when treated with 1% trypan blue solution. The strain was also reisolated from the second calf. The original isolate had been maintained in continuous in vitro cultivation for more than 99 days.

Cryopreservation of Babesia bigemina for in vitro cultivation.

Babesia bigemina-infected RBC and merozoites were cryopreserved and used to initiate in vitro cultures in normal bovine RBC; the cryoprotectant was a final 10% polyvinylpyrrolidone in Vega y Martinez solution. A cooling rate of 20 C/min until -80 C and then rapid transfer to liquid N2 storage was satisfactory. Samples for culture initiation were rapidly thawed at 37 C, washed in Vega y Martinez solution and resuspended in complete culture media containing 10% normal bovine RBC. The optimum culture conditions to reestablish cultures were a 24-well plate (16 mm ID), 5 mm in depth, and an atmosphere of 2% to 5% O2, 5% CO2, and 93% to 90% N2.

Anodic reactions in microbial fuel cells.

Potentiometric and amperometric measurements were made with microbial fuel cells containing E. coli or yeast as the anodic reducing agent and glucose as the oxidizable substrate. The catalytic effects of thionine and resorufin on the anode reaction were investigated. Results on the potentiometry, polarization, and coulombic output of the cells support a mediator-coupled mechanism for the transfer of electrons from the organism to the electrode in preference to a mechanism of "direct" electrochemical oxidation of glucose or its degradation products. Experiments with (14)C-labeled glucose show that when a microbial fuel cell produces a current under load, exogenous glucose is metabolized to produce (14)CO(2). The Coulombic yields of the cells indicate a high degree of energy conversion in these systems.