Venezuelan researchers call for international help.
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Ecology↗
Biomedical subjects
C Mendoza
Publications and source records attributed to C Mendoza.
16K prolactin induces NF-kappaB activation in pulmonary fibroblasts.
The amino-terminal 16 kDa fragment of prolactin (16K PRL) promotes the expression of the inducible isoform of nitric oxide synthase (iNOS) accompanied by the production of nitric oxide (NO) by rat pulmonary fibroblasts. The present study was designed to elucidate whether the mechanism by which 16K PRL promotes iNOS expression involves the activation of nuclear factor-kappa B (NF-kappaB), a key transcription factor for iNOS induction. 16K PRL stimulated DNA-binding activity of NF-kappaB in pulmonary fibroblasts as demonstrated by gel shift assays. Likewise, fluorescence immunocytochemistry showed that 16K PRL promotes nuclear translocation of the p65 subunit of NF-kappaB. Finally, treatment with 16K PRL induced the degradation of the NF-kappaB inhibitor kappaB-beta (IkappaB-beta), and such degradation was prevented by blocking IkappaB-beta phosphorylation. Altogether, these results show that 16K PRL activates NF-kappaB nuclear translocation via the phosphorylation and degradation of IkappaB-beta. These findings are consistent with NF-kappaB being part of the signal transduction pathway activated by 16K PRL to induce iNOS expression.
Active Transport, Cell Nucleus↗
[Recurrent rheumatic fever following prosthetic valve replacement].
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Adult↗
Fertilizable oocytes reconstructed from patient's somatic cell nuclei and donor ooplasts.
The only assisted reproduction treatment now available for women with ovarian failure or irreparable oocyte defects is oocyte donation. However, some women experience psychological barriers to the recourse to donor oocytes, related to the lack of contribution of their proper genes to the progeny. A pilot study in humans suggests that this problem may be overcome by the development of techniques for haploidization of somatic cell nuclei, allowing the formation of new oocytes bearing the complete nuclear genome of the patient. Somatic cell nuclei were obtained from cumulus cells of a patient who failed to produce fertilizable oocytes and were transferred into enucleated oocytes (ooplasts) from a donor. Out of six ooplasts injected with the somatic cell nuclei and fertilized with spermatozoa from the patient's husband, signs of haploidization were detected in three oocytes, two of which subsequently started embryonic development and were cryopreserved for eventual future transfer to the genetic mother. These data show that human oocytes can be used for both reprogramming and haploidization of somatic cell nuclei, allowing reconstruction of genetically own oocytes for patients without, or with seriously disturbed, ovarian function.
Journal Article↗
A fast method to estimate kinetic constants for enzyme inhibitors.
We present a method to determine the reaction type and kinetic constants for enzyme inhibitors that decreases the number of experimental assays by at least a factor of five. It is based on a new theoretical formalism in terms of concentrations that dismisses the requirement of estimating initial velocities. Expressions for the time evolution of the concentrations of all the reactants are also given.
Algorithms↗
A new sterigmatocystin-producing Emericella variant from agricultural desert soils.
An unusual, sterigmatocystin-producing taxon with characteristics of both Emericella nidulans (anamorph Aspergillus nidulans) and Emericella rugulosa (anamorph Aspergillus rugulovalvus, formerly A. rugulosus) was isolated repeatedly during a mycofloral survey of desert cotton field soils where aflatoxin is a chronic problem. Members of this taxon had ascospores with smooth convex walls like E. nidulans but grew slowly like E. rugulosa; moreover, they were similar to an industrial echinocandin B-producing strain which had been classified as "Aspergillus nidulans var. roseus." These new desert isolates were compared with "A. nidulans var. roseus" and representative wild-type isolates of E. nidulans and E. rugulosa using traditional morphological characters, secondary metabolite profiles of mycelial extracts, and Southern blot analysis of genomic DNA. The desert isolates and "A. nidulans var. roseus shared morphological, physiological and molecular characters with E. rugulosa. These isolates constitute a new non-rugulose variant of E. rugulosa.
Agriculture↗
Absorption of iron from unmodified maize and genetically altered, low-phytate maize fortified with ferrous sulfate or sodium iron EDTA.
BACKGROUND: Reducing the phytate content in grains by genetic manipulation is a novel approach to increasing nonheme-iron absorption from mixed diets. Fractional iron absorption from a genetically modified strain of low-phytate maize (LPM) increased significantly, by 50%. OBJECTIVE: We assessed iron absorption from porridges prepared from the same LPM (lpa-1-1 mutant) and unmodified wild-type maize (WTM), both of which were fortified with either ferrous sulfate or sodium iron EDTA. DESIGN: Porridges providing 3.4 mg Fe were fortified with either ferrous sulfate or sodium iron EDTA to provide an additional 1 mg Fe/serving. In 14 nonanemic women, iron absorption was measured as the amount of radioiron incorporated into red blood cells (extrinsic tag method) 12 d after consumption of the study diets. RESULTS: No significant effect of phytate content on iron absorption was found when porridge was fortified with either sodium iron EDTA or ferrous sulfate. Fractional absorption of iron from WTM porridge fortified with sodium iron EDTA (5.73%) was 3.39 times greater than that from the same porridge fortified with ferrous sulfate (1.69%). Fractional absorption of iron from the sodium iron EDTA-fortified LPM porridge (5.40%) was 2.82 times greater than that from LPM porridge fortified with ferrous sulfate (1.91%) (P<0.0001 for both comparisons, repeated-measures analysis of variance). Thus, the previously identified benefit of LPM was no longer detectable when maize porridge was fortified with additional iron. CONCLUSION: Iron was absorbed more efficiently when the fortificant was sodium iron EDTA rather than ferrous sulfate, regardless of the type of maize.
Adult↗
Assisted reproduction with in-vitro-cultured testicular spermatozoa in cases of severe germ cell apoptosis: a pilot study.
BACKGROUND: Apoptosis-related cell damage is known to compromise success rates of assisted reproduction with ejaculated spermatozoa. This study was undertaken to determine whether the frequency of apoptosis-related cell damage and reproductive performance of testicular spermatozoa from men with non-obstructive azoospermia can be improved by in-vitro culture. METHODS: Testicular tissue samples were cultured for 2 days in the presence of 50 IU/l FSH and 1 micromol/l testosterone. The frequency of spermatozoa showing DNA strand breakage and plasma membrane phosphatidylserine externalization was compared in before-culture and after-culture samples. The after-culture samples were used in assisted reproduction attempts. RESULTS: In a group of 11 azoospermic patients with at least two previous intracytoplasmic sperm injection (ICSI) failures, the incidence of DNA strand breakage was high in living testicular spermatozoa from before-culture samples, but significantly lower in after-culture samples (96 versus 30%, P < 0.001). The same applied to the incidence of phosphatidylserine externalization in the motile sperm subpopulation from the before-culture and after-culture samples (83 versus 6%, P < 0.001). Seven ongoing clinical pregnancies (six with fresh embryos and one with cryopreserved embryos) were established. CONCLUSIONS: Severe testicular sperm apoptosis may become a new indication for testicular tissue in-vitro culture before ICSI.
Adult↗
[Trends of etiology of acute bacterial meningitis in Chilean children from 1989 to 1998. Impact of the anti-H influenzae type b vaccine].
BACKGROUND: Acute bacterial meningitis still has a high mortality and rate of complications. AIM: To assess the impact of anti H influenzae vaccination on the epidemiology of acute bacterial meningitis in Chilean children. MATERIAL AND METHODS: A retrospective study of hospital discharge records of patients with acute bacterial meningitis. Causative agents were studied globally, by hospital and by age group. The changes in etiology from 1989 to 1995 were also assessed. Between 1996 and 1998, only those patients with acute bacterial meningitis caused by H influenzae were recollected. RESULTS: In the period prior to vaccination (1989-1995), 1000 cases were registered. The main causative agents were N meningitidis in 33.8%, H influenzae type b in 21.9% and S pneumoniae in 15.4%. The incidence of H influenzae decreased in the period from 36.4 to 9.9% (p < 0.001) and the incidence of N meningitidis increased from 22.9 to 52.1% (p < 0.001). The incidence of S pneumoniae did not change significantly. H influenzae predominated in children between 4 and 24 months of age and N meningitidis predominated in children over 25 months of age. In the period after the introduction of vaccination (1995-1998), there was a further decrease in the incidence of H influenzae from 10 to 2% (p < 0.001). Until 1997, there was a considerable increase in the incidence of N meningitidis, specially in children over 25 months of age. It declined in 1998 to 38%. CONCLUSIONS: There was a reduction in the incidence of acute bacterial meningitis caused by H influenzae prior to the introduction of the vaccine against H influenzae type b. The decrease was more pronounced after the introduction of the vaccine.
Acute Disease↗
Absorption of unlabeled reduced iron of small particle size from a commercial source. A method to predict absorption of unlabeled iron compounds in humans.
The absorption of a commercial brand of small-particle reduced iron was evaluated in 10 normal subjects. For each subject, the hemoglobin incorporation method was used to measure the true absorption of 60 mg of iron from either ferrous sulfate or ferric ammonium citrate. The iron tolerance test (ITT) was also studied for these two compounds and for reduced iron. This procedure consisted of measuring the area under the curve of plasma iron elevations at specified times for 6 hours, or the peak plasma iron, corrected by the plasma iron disappearance rate obtained from measuring plasma iron at specified times for 4 hours after the slow intravenous injection of 0.4 mg of iron as ferric citrate. Only the ITT was used to measure the absorption of 60 mg of reduced iron. Reference dose iron ascorbate absorption was measured in each subject. The absorption of ferric ammonium citrate and reduced iron was expressed as percent of dose and also as absorption percent of that of ferrous sulfate. Mean % geometric "true absorptions" were 39.0 for reference dose, 10.4 for FeSO4 and 2.4 for ferric ammonium citrate. The later was 23% that of FeSO4. By ITT the mean geometric % absorptions were 7.9, 3.7 and 3.2 for FeSO4, ferric ammonium citrate and reduced iron respectively, or 47 and 41% of that of FeSO4. We propose that the true absorption of the commercial brand of reduced iron tested was 20% that of FeSO4 based on the relation between the ITT results of reduced iron and the ITT and true absorption values of ferric ammonium citrate in relation to FeSO4. The use of this method for measuring absorption of unlabeled iron compounds is discussed.
Absorption↗
[Total and fecal coliforms, some enterobacteria staphylococcus sp. and moulds in salads for hot dogs sold in Maracay, Venezuela].
Fast food sold on the streets are good vehicle to transmit gastrointestinal diseases to consumers because they are highly manipulated. The vegetables are a good media for pathogens growth. Raw vegetables salads for hot dog sold in Maracay were studied. Total and fecal Coliforms and Staphylococcus sp. were quantified by the Most Probable Number Technique, Coliforms and Staphylococcus isolated were partially identified. Also the mould population was quantified and identified. Microbial growth was correlated with pH and acidity measurements. MPN/g of Total Coliforms was 1.44 x 10(5) and 4.57 x 10(4) for fecal Coliforms. E. coli was no detected. 87 isolated from the Coliforms were partially identified as Citrobacter freundii variety 1 (45.09% of the isolated), C. freundii variety II (21.57%), Enterobacter aerogenes variety I (17.65%) and E. aerogenes variety II (15.69%); Staphylococcus sp. 3.93 x 10(6) MPN/g and all of its isolated (52) were coagulase negative. The mould population was 4.5 x 10(4) ufc/g, Penicillium sp., Aspergillus, sp., Fusarium sp. were partially identified. The salads presented a pH of 5.92 and an acidity of 0.78 ml of NaOH.
Analysis of Variance↗
In-vitro maturation of immature human male germ cells.
Both meiotic and postmeiotic maturation events have been observed to occur in human male germ cells during in-vitro culture. The temperature of 30 degrees C, medium supplementation with follicle-stimulating hormone and testosterone and the maintenance of the original cell-cell associations within explanted segments of the testicular seminiferous tubules are common features of the most efficient culture systems. The in-vitro maturation processes are markedly accelerated as compared to the in-vivo situation, probably due to the abrogation of a checkpoint controlling the full assembly of molecules needed for spermiogenesis. Moreover, both meiotic and postmeiotic maturation processes can be reactivated in vitro in some cases with a complete block of the same processes in vivo. Healthy babies were born after micromanipulation-assisted fertilization with in-vitro matured elongated spermatids from men with complete in-vivo maturation arrest at the round spermatid stage and at the primary spermatocyte stage.
Cell Differentiation↗
[Knowledge of care activity: another professional factor associated with utilization?].
OBJECTIVES: To analyse the doctors' level of understanding of the use of resources in primary care and the relationship of their understanding to the proper use of resources. SETTING: Primary care in the Autonomous Community of Murcia. DESIGN: Descriptive, crossover study. RESULTS: There was little concordance between the real data contributed by the unit of information and registration of primary care management and data calculated by the doctors themselves. The best-known parameters are frequency of attendance (kappa index = 0.11) and patient pressure (kappa = 0.39). The profile of the doctors did not in general relate to this degree of understanding; it was only observed in a tendency in older doctors (p < 0.001) to overvalue frequency of attendance. The doctors with less understanding used radiology resources more with 164 requests per 1000 inhabitants per year, made more specialist referrals (826 per thousand inhabitants per year) and prescribed more medicines (21 prescriptions per inhabitant per year). Likewise, doctors who used resources most tended to underestimate their use of resources (p < 0.001). CONCLUSIONS: The doctor's understanding of his/her health-care activity and of the resources it involves is scant and independent of the doctor's professional characteristics. This is a factor, which we could call the professional factor, clearly related to resource use.
Clinical Medicine↗
Time-dependent closed form solutions for fully competitive enzyme reactions.
An analytic formalism developed earlier to describe the time evolution of the basic enzyme reaction is extended to fully competitive systems. Time-dependent closed form solutions are derived for the three nominal cases of competition: even, slow and fast inhibitors, allowing for the first time the complete characterization of the reactions. In agreement with previous work, the time-independent Michaelis-Menten approach is shown to be inaccurate when a fast inhibitor is present. The validity of the quasi-steady-state approximation on which the present framework is based is also revised.
Enzyme Inhibitors↗
Birth of healthy twins after fertilization with in vitro cultured spermatids from a patient with massive in vivo apoptosis of postmeiotic germ cells.
OBJECTIVE: To examine whether the results of assisted reproduction with the use of elongated spermatids from a man with incomplete arrest of spermiogenesis and a high frequency of apoptosis among postmeiotic germ cells can be improved by germ cell in vitro culture. DESIGN: Case report. SETTING: Private assisted reproduction centers and a university department. PATIENT(S): Man with incomplete spermiogenesis failure. INTERVENTION(S): Testicular spermatid extraction, in vitro culture of testicular biopsy samples, intraoocyte injection of elongated spermatids, embryo culture, and transfer. MAIN OUTCOME MEASURE(S): Fertilization rate, embryo morphology, and pregnancy. RESULT(S): An assisted reproduction attempt with viability-selected elongated spermatids from a fresh testicular biopsy sample resulted in a poor fertilization result and embryo quality. Further analysis of the sample used in this attempt showed a high incidence of apoptosis among postmeiotic germ cells. A second attempt was then performed with in vitro culture of testicular cells for 24 hours before spermatid injection. This procedure led to a significant decrease in the frequency of apoptosis among viability-selected spermatids, to improvement of the fertilization rate and embryo quality, and to the birth of healthy twins. CONCLUSION(S): In vitro culture of testicular biopsy samples before assisted reproduction with elongated spermatids may improve the efficacy of treatment in cases of massive in vivo apoptosis of postmeiotic germ cells.
Adult↗
The activity (calcium oscillator?) responsible for human oocyte activation after injection with round spermatids is associated with spermatid nuclei.
OBJECTIVE: To compare the oocyte-activating ability of whole human round spermatids and their isolated nuclei. DESIGN: Prospective study using sibling oocytes from patients undergoing spermatid conception and intracytoplasmic sperm injection treatment cycles. SETTING: Private assisted reproduction laboratories and a university department. PATIENT(S): Couples with male infertility. INTERVENTION(S): Sibling oocytes were injected either with whole round spermatids or with their isolated nuclei, followed by artificial triggering of oocyte activation with calcium ionophore. Other sibling oocytes were injected either with isolated spermatid cytoplasm or with whole mature spermatozoa. MAIN OUTCOME MEASURE(S): Numbers of activated oocytes and cleaving embryos. RESULT(S): After oocyte activation was boosted with calcium ionophore, whole spermatids and isolated spermatid nuclei were equally effective in supporting oocyte activation and the formation of pronuclei, whereas no control oocyte was activated under the same conditions after injection of isolated spermatid cytoplasmic compartments. Cleavage rates were lower after the injection of isolated spermatid nuclei than after the injection of whole spermatids. CONCLUSION(S): The factor responsible for human oocyte activation after round spermatid injection is associated with spermatid nuclei. The requirement for the artificial trigger (calcium ionophore) suggests that this factor is identical to the male gamete activity previously characterized as calcium oscillator.
Calcium Signaling↗
Allelotype and loss of heterozygosity around the L-myc gene locus in primary lung cancers.
L-myc S-allele was reported to be associated with metastasis of lung cancer, indicating the existence of a putative tumor suppressor gene around the L-myc locus, in linkage disequilibrium. The relationship between the S-allele and inactivation of some tumor suppressor gene should be indicated by allelic loss. Therefore, we examined the association between the L-myc S-allele and loss of heterozygosity at 11 loci around the L-myc locus (1p34.3) in primary lesions or other biological characteristics in lung cancer. No associations between the S-allele and allelic loss around the L-myc locus or other characteristics were found. According to the deletion map, three shortest regions of overlap between D1S230 and D1S76 were identified. While loss of heterozygosity at SRO1, between D1S2797 and MYCL1, showed no relationship with the pathological stage, it was more frequently observed in squamous cell carcinoma than adenocarcinoma (P=0.019), and associated with high telomerase activity (P=0.046), an indicator of cellular immortality. In conclusion, we found three shortest regions of overlap (SROs) from D1S2797 to pter, and a tumor suppressor gene, which might be associated with suppression of lung cancer development but not with L-myc S-allele, may exist in SRO1.
Adult↗
Immature germ cell conception-in vitro germ cell manipulation.
Experimental studies in laboratory animals have shown that successful conception can be achieved by fertilizing oocytes with immature male germ cells. This gave rise to the concept that immature germ cells recovered from the testes of azoospermic men with maturation arrest may be used for assisted reproduction. However, in contrast to using germ cells recovered from healthy animals, clinical application to the treatment of male sterility is burdened by inherent defects in germ cells attributable to underlying testicular pathology. The recent introduction of in vitro germ cell culture/manipulation techniques makes it possible, in some cases, to overcome the in vivo maturation arrest by allowing an additional meiotic and post-meiotic differentiation and the selective harvesting of cells devoid of apoptosis-related nuclear and cytoplasmic damage. These techniques enabled the first births of normal infants fathered by azoospermic men with maturation arrest at the primary spermatocyte stage and improved the efficacy of assisted reproduction in men with maturation arrest at the round spermatid stage.
Animals↗