Ethnic variation of the C677T and A1298C polymorphisms in the methylenetetrahydrofolate-reductase (MTHFR) gene in southwestern Mexico.

In this study, we examined the distribution of genotype and allele frequencies of the C677T and A1298C polymorphisms in the methylenetetrahydrofolate-reductase gene (MTHFR) in two ethnic groups in the State of Guerrero, Mexico, which were compared with those of the Mestizo population of the region. A comparative study was conducted on 455 women from two ethnic groups and a group of Mestizo women of the State of Guerrero, Mexico: 135 Nahuas, 124 Mixtecas, and 196 Mestizas. Genotyping of both polymorphisms were performed by using polymerase chain reaction-restriction fragment length polymorphism methods. We found that the 677TT genotype was more frequent in Nahua and Mixteca women compared to Mestiza women (P = 0.008), and the most prevalent genotype in both ethnic groups was the 1298AA genotype (P < 0.001). We also compared the 677T allele frequency obtained from the groups studied with the frequencies reported in other ethnic groups of Mexico (Huichol, Tarahumara, and Purepecha). There were significant differences between the three ethnic groups compared to Nahuas (Huicholes, P = 0.004; Tarahumaras, P < 0.001; Purepechas, P = 0.042). Our results indicated significant differences in the frequencies of the C677T and A1298C polymorphisms between the two ethnic groups and the Mestizo population of the State of Guerrero. In addition, we found strong differences with other ethnic groups in Mexico. These results could be useful for future studies investigating diseases related to folate metabolism, and could help the government to design specific nutrition programs for different ethnic groups.

The integration of HR-HPV increases the expression of cyclins A and E in cytologies with and without low-grade lesions.

BACKGROUND: Cyclin-A and cyclin-E are regulators of G1-S phase of normal cell cycle. Integration of human papilloma virus high-risk (HR-HPV) could alter this mechanism, and its overexpression has been associated with poor prognosis in cervical cancer. AIM: To determine the expression of cyclin-A and cyclin-E, types of HR-HPV and physical state of DNA in cytologies with the diagnosis of low-grade squamous intraepithelial lesion (LSIL). MATERIALS AND METHODS: 115 cytological specimens in liquid base (liquid-PREP(™)) were analyzed. 25 specimens were with no signs of SIL (NSIL) and without HPV; 30 with NSIL with low-risk HPV (LR-HPV); 30 with NSIL with HR-HPV; and 30 with both LSIL and HR-HPV. The expression of cyclins was evaluated by immunocytochemistry; and the detection of viral DNA was done by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLPs) for genotyping or sequencing of HPV. The physical state of HPV was evaluated by in situ hybridization with amplification with tyramide. RESULTS: In the cytologies NSIL with LR-HPV, the expression of cyclin-A and cyclin-E was found respectively in 23.3% and 33.3% of the specimens. Among the specimens of NSIL with HR-HPV, 33.3% expressed cyclin-A and 40% cyclin-E, while 100% of the LSILs expressed the 2 cyclins. On the other hand, 100% of the samples NSIL with LR-HPV presented an episomal pattern. Of the specimens of NSIL with HR-HPV, 56.6% exhibited an episomal pattern, 23.3% integrated and 20%, mixed. Among the LSILs, 90% were mixed and 10% integrated. CONCLUSIONS: The cyclins A and E are present in the LSILs that occur predominantly in mixed state in the presence of HR-HPV.

Temporary morphological changes in plus disease induced during contact digital imaging.

OBJECTIVE: To compare and quantify the retinal vascular changes induced by non-intentional pressure contact by digital handheld camera during retinopathy of prematurity (ROP) imaging by means of a computer-based image analysis system, Retinal Image multiScale Analysis. METHODS: A set of 10 wide-angle retinal pairs of photographs per patient, who underwent routine ROP examinations, was measured. Vascular trees were matched between 'compression artifact' (absence of the vascular column at the optic nerve) and 'not compression artifact' conditions. Parameters were analyzed using a two-level linear model for each individual parameter for arterioles and venules separately: integrated curvature (IC), diameter (d), and tortuosity index (TI). RESULTS: Images affected with compression artifact showed significant vascular d (P<0.01) changes in both arteries and veins, as well as in artery IC (P<0.05). Vascular TI remained unchanged in both groups. CONCLUSIONS: Non-adverted corneal pressure with the RetCam lens could compress and decrease intra-arterial diameter or even collapse retinal vessels. Careful attention to technique is essential to avoid absence of the arterial blood column at the optic nerve head that is indicative of increased pressure during imaging.

Retinopathy of prematurity as a major cause of severe visual impairment and blindness in children in schools for the blind in Guadalajara city, Mexico.

AIM: To determine the causes of blindness in students attending schools for the blind in Guadalajara city, Mexico and to assess the availability of screening for retinopathy of prematurity (ROP) in local neonatal intensive care units. METHODS: Information on causes of blindness was obtained by interview with parents and teachers, review of records and examination. Causes of visual loss in children with a distance visual acuity of <6/60 (ie, severely visually impaired or blind) were determined and classified according to the WHO's classification system for children. RESULTS: Of 153 children in the two participating schools, 144 were severely visual impaired or blind. Their ages ranged from 4 months to 15 years and 58% were female. ROP was the most common cause of visual loss (34.7%), followed by optic nerve lesions (17.4%) and glaucoma (14.6%). 25/59 (42.3%) children aged 0-4 years were blind from ROP compared with 6/32 (18.8%) children aged 10-15 years. 78% of children blind from ROP had psychomotor delay and less than half (46%) had not received treatment for ROP. All five privately funded neonatal intensive care units in the city regularly screen for ROP compared with only four of the 12 units in the public sector. CONCLUSIONS: ROP is the leading cause of blindness in children in Mexico despite national guidelines being in place. Health policies promoting primary prevention through improved neonatal care need to be implemented. Advocacy is required so that the time ophthalmologists spend screening and treating ROP is included in their job description and hence salaried.

Evaluation of the metal phytoextraction potential of crop legumes. Regulation of the expression of O-acetylserine (thiol)lyase under metal stress.

The metal phytoextraction potential of three legumes belonging to different genera has been studied under greenhouse conditions. Legumes accumulate As and metals mainly in roots, although translocation to shoot is observed. Alfalfa did accumulate the highest concentrations of As and metals in shoots and aerial biomass was less affected by the toxic elements, indicating its good behaviour in phytoextraction. Clover accumulated less metal, but showed larger biomass. EDTA addition enhanced Pb phytoextraction up to levels similar to those described for plants proposed in phytoremediation. The regulation of O-acetylserine (thiol)lyase from legumes under metal stress has been analysed to test the possibility of establishing a possible correlation between the expression of OASTL in the presence of the metals and the metal accumulation in legume plant tissues. Cd and Pb(EDTA) produce the strongest increases of OASTL activity, with the higher enhancement seen in roots, in parallel with the higher metal accumulation. Arsenic produced an increase of root enzyme activity, whereas Cu produced a decrease, mainly in shoots. Western blots using antibodies against an A. THALIANA cytosolic OAS-TL recognised up to five protein bands in crude extracts from LOTUS and clover. A low molecular weight isoform of 32 kDa was induced in the presence of Cd and Pb. A partial RT-PCR sequence from clover has been obtained, showing 86 - 97 % identity with other described OASTLs. The PCR fragment has been used to analyse OASTL mRNA levels of legumes under metal stress. OASTL transcripts were increased by As, Cd, and Pb, especially in roots, where metal accumulation was maximal, while Cu produced a decrease in the transcript levels.

Orbital angular momentum exchange in the interaction of twisted light with molecules.

In the interaction of molecules with light endowed with orbital angular momentum, an exchange of orbital angular momentum in an electric dipole transition occurs only between the light and the center of mass motion; i.e., internal "electronic-type" motion does not participate in any exchange of orbital angular momentum in a dipole transition. A quadrupole transition is the lowest electric multipolar process in which an exchange of orbital angular momentum can occur between the light, the internal motion, and the center of mass motion. This rules out experiments seeking to observe exchange of orbital angular momentum between light beams and the internal motion in electric dipole transitions.

The cytosolic O-acetylserine(thiol)lyase gene is regulated by heavy metals and can function in cadmium tolerance.

Regulation of the expression of the cytosolic O-acetylserine(thiol)lyase gene (Atcys-3A) from Arabidopsis thaliana under heavy metal stress conditions has been investigated. Northern blot analysis of Atcys-3A expression shows a 7-fold induction after 18 h of cadmium treatment. Addition of 50 microm CdCl(2) to the irrigation medium of mature Arabidopsis plants induces a rapid accumulation of the mRNA throughout the leaf lamina, the root and stem cortex, and stem vascular tissues when compared with untreated plants, as observed by in situ hybridization. High pressure liquid chromatography analysis of GSH content shows a transient increase after 18 h of metal treatment. Our results are compatible with a high cysteine biosynthesis rate under heavy metal stress required for the synthesis of GSH and phytochelatins, which are involved in the plant detoxification mechanism. Arabidopsis-transformed plants overexpressing the Atcys-3A gene by up to 9-fold show increased tolerance to cadmium when grown in medium containing 250 microm CdCl(2), suggesting that increased cysteine availability is responsible for cadmium tolerance. In agreement with these results, exogenous addition of cystine can, to some extent, also favor the growth of wild-type plants in cadmium-containing medium. Cadmium accumulates to higher levels in leaves of tolerant transformed lines than in wild-type plants.

Glutathione biosynthesis in Arabidopsis trichome cells.

In Arabidopsis thaliana, trichome cells are specialized unicellular structures with uncertain functions. Based on earlier observations that one of the genes involved in cysteine biosynthesis (Atcys-3A) is highly expressed in trichomes, we have extended our studies in trichome cells to determine their capacity for glutathione (GSH) biosynthesis. First, we have analyzed by in situ hybridization the tissue-specific expression of the genes Atcys-3A and sat5, which encode O-acetylserine(thio)lyase (OASTL) and serine acetyltransferase (SAT), respectively, as well as gsh1 and gsh2, which encode gamma-glutamylcysteine synthetase and glutathione synthetase, respectively. The four genes are highly expressed in leaf trichomes of Arabidopsis, and their mRNA accumulate to high levels. Second, we have directly measured cytoplasmic GSH concentration in intact cells by laser-scanning microscopy after labeling with monochlorobimane as a GSH-specific probe. From these measurements, cytosolic GSH concentrations of 238+/-25, 80+/-2, and 144+/-19 microM were estimated for trichome, basement, and epidermal cells, respectively. Taking into account the volume of the cells measured using stereological techniques, the trichomes have a total GSH content more than 300-fold higher than the basement and epidermal cells. Third, after NaCl treatment, GSH biosynthesis is markedly decreased in trichomes. Atcys-3A, sat5, gsh1, and gsh2 mRNA levels show a decrease in transcript abundance, and [GSH](cyt) is reduced to 47+/-5 microM. These results suggest the important physiological significance of trichome cells related to GSH biosynthesis and their possible role as a sink during detoxification processes.

[Activities of the Brazilian Senate in the area of public health, 1995 and 1996]. / Atividades do Senado Federal brasileiro na área de saúde pública, 1995 e 1996.

The objective of this study was to identify and describe the actions of Brazilian senators in the field of public health in 1995 and 1996. We also sought to determine if profession, regional background, or political party influenced the senators' actions. The actions were divided into three types: legislative (proposal and review of bills and petitions); supervisory (information requests to the executive branch and the establishment of inquiry committees and other special committees); and parliamentary (speeches). The data were collected from two databases maintained by the Senate, namely MATE and DISC. Of the 89 senators who were in office during the study period, 76 were involved with public health issues. Of the total of 667 actions studied, there was a predominance of speeches (43% of all actions), most of them responding to news reported by regional or national media. Supervisory activities were limited (5% of all actions were information requests). The subjects dealt with most frequently were health policies (30%), drugs (9%), regulation of health professions (8%), disease control (7%), and worker health (6%). Concerning the professions of the senators, the most frequent categories were physicians, teachers, and journalists. The senators representing the North and Northeast regions performed 62% of the actions and were involved with almost all the health subjects. Although 43% of the actions were carried out by liberal and right-wing senators, the senators from socialist and labor parties had a stronger proportional participation (both in terms of senators involved and actions performed). It is interesting to note that socialist and labor senators showed minimal involvement in the issue of worker health. The predominance of speeches as a prevalent type of action, the limited and disjointed scope of legislative actions, and, especially, the poor monitoring and control show the need for deep changes in the work process of the Senate's technical committees and, therefore, of the Senate's technical support unit.

Large epidemics of hemorrhagic fevers in Mexico 1545-1815.

In 1545, twenty-four years after the Spanish conquest of the Aztec empire, an epidemic of a malignant form of a hemorrhagic fever appeared in the highlands of Mexico. The illness was characterized by high fever, headache, and bleeding from the nose, ears, and mouth, accompanied by jaundice, severe abdominal and thoracic pain as well as acute neurological manifestations. The disease was highly lethal and lasted three to four days. It attacked primarily the native population, leaving the Spaniards almost unaffected. The hemorrhagic fevers remained in the area for three centuries and the etiologic agent is still unknown. In this report we describe, and now that more information is available, analyze four epidemics that occurred in Mexico during the colonial period with a focus on the epidemic of 1576 which killed 45% of the entire population of Mexico. It is important to retrieve such diseases and the epidemics they caused from their purely historical context and consider the reality that if they were to reemerge, they are potentially dangerous.

Salt-specific regulation of the cytosolic O-acetylserine(thiol)lyase gene from Arabidopsis thaliana is dependent on abscisic acid.

The expression of Atcys-3A gene coding for cytosolic O-acetylserine(thiol)lyase, a key enzyme in cysteine biosynthesis, from Arabidopsis thaliana is significantly induced by exposure to salt and heavy-metal stresses. Addition of NaCl to mature plants induced a rapid accumulation of the mRNA throughout the leaf lamina and roots, and later on in stems, being mainly restricted to vascular tissues. The salt-specific regulation of Atcys-3A was also mediated by abscisic acid (ABA) since: (1) exogenous addition of ABA to the culture medium mimicked the salt-induced plant response by raising the level of Atcys-3A transcript, and (2) Arabidopsis mutants aba-1 and abi2-1 were not able to respond to NaCl. Our results suggest that a high rate of cysteine biosynthesis is required in Arabidopsis under salt stress necessary for a plant protection or adaptation mechanism. This hypothesis was supported by the observation that intracellular levels of cysteine and glutathione increased up to 3-fold after salt treatment.

Isolation and analysis of the soybean SGA2 gene (cDNA), encoding a new member of the plant G-protein family of signal transducers.

We have isolated a cDNA clone from Glycine max, named SGA2, coding for a G alpha-subunit protein. The encoded polypeptide, SG alpha2, shows a molecular mass of 45 kDa and contains most of the conserved regions involved in guanine nucleotide binding and hydrolysis. Comparison at the nucleotide and amino acid sequence levels with the other plant G alpha's shows a high degree of conservation (>85% similarity). Phylogenetic analysis of these plant genes with the other G alpha's from different species clearly indicate that those proteins represent a new member of the heterotrimeric G-protein family, named Gp. Tissue localization of SGA2 transcripts in root, stem and leaf organs shows that this gene is widely expressed throughout the plant although it is most abundant in the vascular tissues of all these organs. Furthermore, the transcript is more abundant in young tissues and organ primordia than mature tissues. The high degree of sequence conservation among the plant G alpha's and the differences to other species of other kingdoms, suggest that plant G proteins may function in specialized signalling processes.

A novel endo-beta-1,3-glucanase, BGN13.1, involved in the mycoparasitism of Trichoderma harzianum.

The mycoparasitic fungus Trichoderma harzianum CECT 2413 produces at least three extracellular beta-1,3-glucanases. The most basic of these extracellular enzymes, named BGN13.1, was expressed when either fungal cell wall polymers or autoclaved mycelia from different fungi were used as the carbon source. BGN13.1 was purified to electrophoretic homogeneity and was biochemically characterized. The enzyme was specific for beta-1,3 linkages and has an endolytic mode of action. A synthetic oligonucleotide primer based on the sequence of an internal peptide was designed to clone the cDNA corresponding to BGN13.1. The deduced amino acid sequence predicted a molecular mass of 78 kDa for the mature protein. Analysis of the amino acid sequence indicates that the enzyme contains three regions, one N-terminal leader sequence; another, nondefined sequence; and one cysteine-rich C-terminal sequence. Sequence comparison shows that this beta-1,3-glucanase, first described for filamentous fungi, belongs to a family different from that of its previously described bacterial, yeast, and plant counterparts. Enzymatic-activity, protein, and mRNA data indicated that bgn13.1 is repressed by glucose and induced by either fungal cell wall polymers or autoclaved yeast cells and mycelia. Finally, experimental evidence showed that the enzyme hydrolyzes yeast and fungal cell walls.

Analysis of three tissue-specific elements from the wheat Cab-1 enhancer.

The genes encoding the major light-harvesting antenna chlorophyll a/b-binding protein (Cab) of higher plants are regulated by light at the transcriptional level. In addition, their expression is largely restricted to photosynthetically competent organs such as leaves. A 268 bp fragment of the Cab-1 promoter from wheat functions as a light-responsive and organ-specific enhancer in transgenic tobacco. Using DNase I footprinting, four different regions have been mapped (Cab1-A, Cab1-B, Cab1-C and Cab1-D) in this fragment that bind to protein factors in tobacco nuclear extracts. Three of these regions (A, B and C) coincide with sequences that have been found to be functionally important from previous cis-element analyses. Synthetic tetramers of these three sites interact with different proteins in gel retardation assays. In addition, cross-competition analyses demonstrate that Cab1-C is likely to interact with ASF-2, a tobacco DNA-binding activity that binds to a conserved GATA element found in many dicot Cab promoters. In transgenic tobacco, a 95 bp fragment of the Cab-1 enhancer containing the A, B and C regions can confer leaf expression when fused upstream of a truncated derivative of the cauliflower mosaic virus (CaMV) 35S promoter. However, expression observed with this enhancer fragment in the promoter context of these studies does not appear to be significantly dependent on light. Similar results were obtained with synthetic tetramers of Cab1-A, -B or -C. These data thus suggest that the wheat Cab-1 enhancer contains at least three distinct elements that contribute to leaf-specific expression in transgenic tobacco. Interaction between factors binding to these positive elements and those that bind elsewhere in the Cab-1 enhancer may be necessary for light-responsive transcriptional activation.

Guanine nucleotide binding protein involvement in early steps of phytochrome-regulated gene expression.

The transmission process of light signals from plant photoreceptors to target cellular events is largely unknown. In the present work, we show that treatment of dark-adapted soybean cells (SB-P) with cholera toxin or pertussis toxin uncouples phytochrome-dependent gene expression. Addition of as little as 10 ng of toxin per ml is sufficient to activate expression of genes encoding the major chlorophyll a/b-binding protein (cab) in the dark. Significant levels of cab transcript accumulation are detected within 0.5 h after addition of the toxins and expression of these genes is desensitized to further light treatments. Treatment of SB-P cells with the calmodulin antagonist N-(6-aminohexyl)-5-chloro-1-naphtha-lenesulfonamide hydrochloride (W-7) prevents induction of the photoregulated gene by phytochrome or bacterial toxins. These results indicate the involvement of guanine nucleotide binding protein(s) in phytochrome-mediated cab gene activation. A likely site of action for this step is between the photoreceptor and a downstream W-7-sensitive effector.