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Mitochondrial DNA (mtDNA) methylation has the potential to be used as a biomarker of human development or disease. However, mtDNA methylation procedures are costly and time-consuming. Therefore, we developed a new approach based on an RT-PCR assay for the base site identification of methylated cytosine in the control region of mtDNA through a simple, fast, specific, and low-cost strategy. Total DNA was purified, and methylation was determined by RT-PCR bisulfite sequencing. This procedure included the DNA purification, bisulfite treatment and RT-PCR amplification of the control region divided into three subregions with specific primers. Sequences obtained with and without the bisulfite treatment were compared to identify the methylated cytosine dinucleotides. Furthermore, the efficiency of C to U conversion of cytosines was assessed by including a negative control. Interestingly, mtDNA methylation was observed mainly within non-Cphosphate- G (non-CpG) dinucleotides and mostly in the regions containing regulatory elements, such as OH or CSBI, CSBII, and CSBIII. This new approach will promote the generation of new information regarding mtDNA methylation patterns in samples from patients with different pathologies or that are exposed to a toxic environment in diverse human populations.

Sex identification of ancient individuals is important to understand aspects of the culture, demographic structure, religious practices, disease association, and the history of the ancient civilizations. Sex identification is performed using anthropometric measurements and molecular genetics techniques, including quantification of the X and Y chromosomes. These approaches are not always reliable in subadult, or fragmented, incomplete skeletons or when the DNA is highly degraded. Most of the methods include the identification of the male and female sexes, but the absence of a specific marker for the males does not mean that the sample obtained was from a female. This study aims (1) to identify new male-specific regions that allow male identification; (2) to contrast the effectiveness of these markers against AMELX/AMELY and anthropometric measurement procedures; and (3) to test the efficacy of these markers in archaeological samples. For the first two aims, we used known sex samples, and for the third aim, we used samples from different archaeological sites. A novel molecular technique to identify male-specific regions by amplification of TTTY7, TSPY3, TTTY2, and TTTY22 genes of the human Y chromosome was developed. The results showed amplification of the specific DNA regions of Y chromosome in male individuals, with no amplification being observed in any of the female samples, confirming their specificity for male individuals. This approach complements the current procedures, such as the AMELX/AMELY test and anthropometric principle.

Previous data have established direct archaeological evidence that semisedentary forager-farmers in the Mesoamerican region existed during the millennia preceding the onset of sedentary agrarian life in the Maya area. The present study concerned the identification of the maternal genetic origin of skeletal remains discovered in the Puyil cave through sequence analysis of the mitochondrial DNA. The individuals pertained to the Archaic and Classic periods and the samples were found inside the Puyil cave located in Tabasco, Mexico, an area occupied by Zoque and Maya populations. The technologies used were next-generation sequencing and Sanger sequencing. The analysis was was made possible by the development of haplotype networks for comparing mitochondrial DNA sequences from ancient and contemporary civilizations from the American continent and Asia. The sequences displayed haplogroups A, A2, C1, C1c, and D4, while haplogroup B was absent. The haplotype networks showed a close genetic relationship with Maya populations from Mexico and Guatemala and with contemporary native populations from Bolivia, Brazil, Dominican Republic, China, and Asia. The analysis with ancient mitochondrial DNA study sequences showed evidence of Maya ancestors’ migration from North to South America and through the Caribbean islands. The results of this study confirm that a Maya ancestor of the ancient remains from the Puyil cave is shared with contemporary Maya and that expansion of the founding haplogroups A, C, and D came from Asia.

Mercury (Hg) is a highly toxic element that causes bone defects and malformations. Structure and surface analyses using quantitative x-ray diffraction using the Rietveld method, High-Resolution Transmission Electron Microscopy and nanodiffraction analyses, and Fourier-Transformed Infrared spectroscopy showed that bone enriched naturally with Hg (≤ 2.3 %) contained Hg3PO4 [(Hg2)3(PO4)2] and HgO. Bone [mostly as apatite, verified as carboxyapatite Ca10(PO4)4(CO3)3(OH)2(s)] and cinnabar (HgS) dissolved releasing Hg+ (existing as dimer Hg22+) and PO43−, both of which became immobilized as (Hg2)3(PO4)2. Besides, released Hg2+ became oxidized to form HgO. The outcome of this work is novel, provided that only a handful of stable compounds of Hg22+ are found in nature.

Dengue virus (DENV) evolution has had a significant impact on disease pathogenesis, virulence, and epidemiology in Mexico. Novel genotypic variation in DENV serotypes and genotypes may influence the magnitude and severity of dengue epidemics, as evidenced by 2009 data from Veracruz State. The data presented herein is related to the publication entitled "Epidemiological Implications of the Genetic Diversification of Dengue Virus (DENV) Serotypes and Genotypes in Mexico" [1]. Raw data and trees provide epidemiological data on DENV prevalence and a comprehensive phylogeny of both representative sequences collected from an NCBI repository, and 28 additional isolates from acute-phase plasma samples diagnosed with dengue fever or severe dengue (Raw sequencing data is hosted in the public repository Mendeley Data (http://dx.doi.org/10.17632/bf2kdhhf6x.2). Phylogenetic trees for each DENV serotype (DENV-1, -2, -3 and -4) were constructed using these sequences by a maximum likelihood methodology as well as a Bayesian Markov chain Monte Carlo (MCMC) integration approach. Phylogenetic trees exhibited: (1) DENV-1, genotype V, (2) the DENV-2 Asian/American and Asian II genotypes, (3) DENV-3, genotype III, and (4) DENV-4, genotype I. This data can be beneficial for future analyses on DENV serotype and genotype structure and the introduction of novel DENV genotype sequences in the Americas, for the further elucidation of dengue etiology.

Introduction Genetic variants have been replicated for association with type 2 diabetes mellitus (T2D) and many of them with diabetes-related traits. Because T2D is highly prevalent in Mexico, this study aimed to test the association of CDKN2A/B, PPARGC1A, VEGFA, SIRT1 and UCP2 gene polymorphisms (rs10811661, rs8192678, rs2010963, rs7896005 and rs659366 respectively) with metabolic traits in 415 unrelated Mexican mestizos with T2D under three models of inheritance. Material and methods A total of 415 unrelated Mexican mestizos were genotyped by TaqMan assays. Triglycerides, cholesterol, glucose, high-density lipoprotein cholesterol (HDL-C), insulin and anthropometric measurements were determined and the HOMA-IR was calculated. Association studies were tested by the Kruskal-Wallis test, linear regression, statistical power analysis, Bonferroni correction, paired SNP analysis, and physical interaction by GeneMANIA. Results All polymorphisms were in Hardy-Weinberg equilibrium, and the association by genotype with T2D-related traits displayed nominal significance for rs8192678 with glucose (p = 0.023) and triglycerides (p = 0.013); rs2010963 with diastolic blood pressure (DBP) (p = 0.012) and cholesterol (p = 0.013); rs7896005 with DBP (p = 0.012) and insulin (p = 0.011); and rs659366 with cholesterol (p = 0.034), glucose (p = 0.031) and triglycerides (p = 0.028); and the association of rs2010963 with HDL-C (p = 0.0007) was significant. Linear regression performed with three models of inheritance, adjusted by age + sex + BMI and corrected with Bonferroni, showed a significant association of rs2010963 with HDL-C in an additive model (p = 0.007); and rs7896005 was significantly associated with DBP in the recessive model (p = 0.006). Conclusions Rigorous analysis evidenced the association of VEGFA rs2010963 and SIRT1 rs7896005 with HDL-C and DBP respectively; these traits are known predictors of cardiovascular complications, which increase the risk of cardiovascular diseases in this population.

Archaeological bone remains are exposed to depositional conditions that profoundly affect their preservation. This manuscript reports on the mineral transformation, chemical composition, and preservation of organic material in pre-Hispanic archaeological Hg-enriched bone remains exposed to different depositional conditions, namely, open and confined environments. Here, we studied 2 vertebrae specimens collected inside Temple XIII of the archaeological site of Palenque, Mexico. Specifically, the 2 vertebrae were collected inside and outside a sarcophagus (VIS and VOS) located in a burial chamber and belonged to a 30-yr-old woman and an 11-yr-old boy, respectively. Subsections of the vertebrae specimens were characterized by radiocarbon dating, high-resolution electron microscopy (HRSEM), and energy-dispersive X-ray spectroscopy analyses (EDS). VIS and VOS showed conventional radiocarbon ages dating of 5050 ± 30 BP and 5440 ± 30 BP respectively. HRSEM-EDS analysis for VIS showed a high content of Hg ( ~ 30%) in the upper layer and below the detection limit in the lower layer. In contrast, HRSEM-EDS analysis for VOS confirmed the lack of a layered structure, for which the accumulation of Hg was as high as 30%. Furthermore, VIS was the only vertebrae that showed the presence of S. This was because VIS belonged to a pre-Hispanic character exposed to HgS, typically used in Mayan royal rituals; VOS did not contain S because the Hg source in this case was probably from cooking utensils, the elaboration of paintings or murals, or the ingestion of contaminated food or water. In addition, VOS contained a particularly low content of P, which was attributed to hydroxyapatite undergoing higher rates of transformation in the open environment. Moreover, the incorporation of Hg in hydroxyapatite via Hg-for-Ca substitution further accelerated the mineral transformation pathway(s). Finally, the pre-Hispanic archaeological Hg-enriched bones preserved the genetic material, regardless of the depositional environments (exposed or unexposed). Therefore, the prevailing preservation mechanisms differ from those underpinned by physical protection by collagen fibrils or natural antibacterial mechanisms.

Variation and clade shifts in dengue virus (DENV) genotypes are responsible for numerous dengue fever outbreaks throughout Latin America in the past decade. Molecular analyses of dengue serotypes have revealed extensive genetic diversification and the emergence of new genotypes in Brazil (DENV-4 genotype I) and elsewhere in tropical and subtropical America. The goal of the present study is to assess the extent to which the adventitious introduction of DENV genotypes and their increasing genetic diversity affects dengue epidemiology in Mexico. A nuanced sequence inspection and phylogenetic analysis of the C-prM nucleotide region of DENV was performed for specimens collecting in 2009 from the Veracruz State, Mexico. Findings were contrasted with specimens collected in adjacent years and analysed based on the epidemiological patterns reported between 1990 and 2019. Additionally, the identification process of various DENV genotypes was assessed, including: (1) DENV-1, genotype V, (2) the DENV-2 Asian/American and Asian II genotypes (3) DENV-3, genotype III, and (4) DENV-4, genotype I. This resulted in the discovery of a distinct genetic cladistic pattern for serotype DENV-2. Lastly, study findings suggest that a correlation exists between the emergence of novel genotypes and genetic diversification, with the increasing incidence of DENV infections in Mexico in 2009.

Mexican Maya populations have a notably high prevalence of type 2 diabetes (T2D) as a consequence of the interaction between environmental factors and a genetic component. To assess the impact of 24 single nucleotide variants (SNVs) located in 18 T2D risk genes, we conducted a family-based association evaluation in samples from Maya communities with a high incidence of the disease. A total of four hundred individuals were recruited from three Maya communities with a high T2D incidence. Family pedigrees (100) and 49 nuclear families were included. Genotyping was performed by allelic discrimination with TaqMan probes. This study also included the family-based association test (FBAT) statistic U to assess the genetic associations with T2D, and the multivariate statistical and haplotype analyses. A positive association with TD2 risk was found for WFS1 rs6446482 (p = 0.046, Z = 1.994) under an additive model, and SIRT1 rs7896005 (p = 0.038, Z = 2.073) under the dominant model. Multivariate model analysis, including T2D status, age, and body mass index (BMI), displayed significant covariance in PPARGC-1α rs8192678; SIRT1 rs7896005; TCF7L2 rs7903146 and rs122243326; UCP3 rs3781907; and HHEX rs1111875 with a P 0.05. This study revealed an association of SIRT1 and WFS1 with T2D risk.

Mitochondrial DNA (mtDNA) copy number and mitochondrial DNA haplogroups have been associated with different types of cancer, including breast cancer, because they alter cellular energy metabolism. However, whether mtDNA copy number or haplogroups are predictors of oxidative stress-related risks in human breast cancer tissue in Mexican patients remains to be determined. Using quantitative real-time PCR assays and sequencing of the mtDNA hypervariable region, analysis of mtDNA copy numbers in 82 breast cancer tissues (BCT) and matched normal adjacent tissues (NAT) was performed to determine if copy number correlated with clinical features and Amerindian haplogroups (A2, B2, B4, C1 and D1) . The results showed that the mtDNA copy number was significantly decreased in BCT compared with NAT (p = 0.010); it was significantly decreased in BCT and NAT in women 50 years of age, compared with NAT in women 50 years of age (p = 0.032 and p = 0.037, respectively); it was significantly decreased in NAT and BCT in the postmenopausal group and in BCT in the premenopausal group compared with NAT in the premenopausal group (p = 0.011, p = 0.010 and, p = 0.018; respectively); and it was also significantly decrease in members of the BCT group classified as having invasive ductal carcinoma I-III (IDC-I, IDC-II and IDC-III) and IDC-II for NAT compared to IDC-I of NAT (p = 0.025, p = 0.022 and p = 0.031 and p = 0.020; respectively). The mtDNA copy number for BCT from patients with haplogroup B2 was decreased compared to patients with haplogroup D1 (p = 0.01); for BCT from patients with haplogroup C1 was also decreased compare with their NAT counterpart (p = 0.006) and with BCT patients belonging to haplogroups A2 and D1 (p = 0.01 and p = 0.03; respectively). In addition, the mtDNA copy number was decrease in the sequences with three deletions relative to the rCRS at nucleotide positions A249del, A290del and A291del, or C16327T polymorphism with the same p = 0.019 for all four variants. Contrary, the copy number increased in sequences containing C16111T, G16319A or T16362C polymorphisms (p = 0.021, =0.048, and = 0.001; respectively). In conclusion, a decrease in the copy number of mtDNA in BCT compared with NAT was shown by the results, which suggests an imbalance in oxidative phosphorylation (OXPHOS) that can affect the apoptosis pathway and cancer progression. It was also observed an increase of the copy number in samples with specific polymorphisms, which may be a good sign of favourable prognosis.

Mitochondria both produce the energy of the cell as ATP via respiration and regulate cellular metabolism. Accordingly, any deletion or mutation in the mitochondrial DNA (mtDNA) may result in a disease. One of these diseases is Kearns Sayre syndrome (KSS), described for the first time in 1958, where different large-scale deletions of different sizes and at different positions have been reported in the mitochondrial genome of patients with similar clinical symptoms. In this study, sequences of the mitochondrial genome of three patients with clinic features of KSS were analyzed. Our results revealed the position, heteroplasmy percentage, size of deletions, and their haplogroups. Two patients contained deletions reported previously and one patient showed a new deletion not reported previously. These results display for the first time a systematic analysis of mtDNA variants in the whole mtDNA genome of patients with KSS to help to understand their association with the disease.

Type 2 diabetes mellitus (T2D) is one of the two leading causes of mortality in Mexico. However, most studies have focused on Caucasians or Asians, and there are a small number of studies investigating Maya populations. Furthermore, to the best of our knowledge, there is no information on isolated Maya communities with T2D frequencies of 20% that are primarily attributed to ethnicity. Consequently, this study focused on assessing which genetic risk variants could be involved in the high rates of T2D in 92 individuals with Maya ancestry; 47 were diagnosed with T2D, and 45 were classified as healthy individuals. A pilot genome-wide association study was performed using the Affymetrix Axiom Genome-wide LAT1 array. The population structure was determined with the ADMIXTURE software using 1289 Latin American selected polymorphisms, and 39 polymorphisms associated with T2D were included for replication. Association tests were performed using the Statistical Analysis System (SAS) using the allelic, genotype and Armitage trend tests. The results indicated that population structure analysis displayed no differences between T2D patients and healthy individuals; 24 loci located were identified for probable association with T2D (p > 1.288 × 10−7 and p

Type 2 diabetes (T2D) is a disease with a prevalence of 9.4% in Mexicans. Its etiology is complex involving environmental and genetic factors. The aim of this study was to analyse the association between PPARG rs1801282, PPARGC1A rs8192678, VEGFA rs2010963, ADRA2A rs553668, KCNQ1 rs2237892, SIRT1 rs7896005, IGF2BP2 rs4402960, and UCP3 rs3781907 single nucleotide variants (SNVs) with T2D and metabolic traits in a case-control study of a population from Mexico City. A total of 831 blood samples of non-diabetic, with healthy control participants (416) and individuals with T2D (415) were collected over a five-year period. After DNA extraction, genotyping was performed with TaqMan probes using real-time PCR. The genotypes were analysed for association with T2D in linear and logistic regressions adjusting for age, sex, and body mass index using the dominant, recessive, and additive models with a Bonferroni correction for multiple comparisons p 0.001 and for association with related T2D traits fixed with a p 2.3 × 10

Maya communities have been shown to exhibit type 2 diabetes (T2D) with high prevalence compared with Mexican mestizo populations. Furthermore, some variants associated with the risk for T2D have been described. In this study, we describe the results of a pilot genome wide association study (GWAS) using 817,823 single nucleotide polymorphisms (SNPs) to identify candidate variants for replication in future studies. Herein, we present the GWAS study data, which were divided into three parts: first, 1289 ancestry informative markers (AIMs) were selected for Latino populations containing European, African, and Native American SNPs obtained from the literature; second, a GWAS hypothesis free to select candidate genes associated with T2D was performed, which identified 24 candidate genes; and third, 39 SNPs previously associated with T2D or related traits were replicated. This article is associated with the original article published in “Gene” under the title “Pilot genome-wide association study identifying novel risk loci for type 2 diabetes in a Maya population”.

Here; we have described and tested a microarray based-method for the screening of dengue virus (DENV) serotypes. This DNA microarray assay is specific and sensitive and can detect dual infections with two dengue virus serotypes and single-serotype infections. Other methodologies may underestimate samples containing more than one serotype. This technology can be used to discriminate between the four DENV serotypes. Single-stranded DNA targets were covalently attached to glass slides and hybridised with specific labelled probes. DENV isolates and dengue samples were used to evaluate microarray performance. Our results demonstrate that the probes hybridized specifically to DENV serotypes; with no detection of unspecific signals. This finding provides evidence that specific probes can effectively identify single and double infections in DENV samples.

The Red Queen remains (700 A.C.) found at Palenque, Mexico, are examples of cinnabar (HgS) application to royal remains during pre-Hispanic times. The Red Queen remains are those of a ca. 30–35-yr-old female and present a striking similarity to the remains of another Mayan woman found at Copan, Honduras. Thus, covering the remains of royal women with HgS may have been a common practice in the Mayan civilization. High resolution microdiffraction and microscopic analysis of the Red Queen remains showed the presence of nanotubular organic minerals comparable in composition and molecular dimensions to collagen fibrils, and in spatial ordering to collagen fiber networks. Fungal structures are rare in the geological record because of poor preservation potential. Micrographs revealed the preservation of fungal signatures, with morphology comparable to parasitic fungal-coral matrix associations, consistent with the idea that fungal remains can be preserved in environments which contain high Hg concentrations. The...

In this paper we use Scanning and Transmission Electron Microscopy, and Energy Dispersive Spectroscopy to characterize the surface of the skin of a 2300 YO, naturally-preserved mummy, belonging to a two-year and eight month girl (“ Pepita ”), found inside a cave located in Altamira, Queretaro, Mexico (21°40′–20°01′; 99°03′–100°36′). The cave was found in Sierra Gorda, an orographic region with a relief of sedimentary origin from the Gulf of Mexico, composed by high mountains with altitude values surpassing 3000 m above sea level, with ample and steep canyons, and a prominent role on the exploitation and distribution of cinnabar (HgS). The skin showed the presence of small-sized spherules, containing Al (≤43%) in the most exposed region (5–10 μm depth). Thin layers and structural microdomains covered small and large spheres. Structures conformed by stacked, nano-sized particles located far-from-the bunches contained C (≤45%), Zn and Si (≤10%), and minor amounts of Ca (≤2.6%). By contrast, regions between spheres contained high amounts of Ca (≤23%) and Al (≤15%), but lacked Zn and Si. Carbon spheres showed two distinctive composition, a signature that their formation might have occurred in a least two different stages via concentric growth mechanisms, with the incorporation of Zn and Al at a later stage. [Zn][Al]Carbon spheres showed morphology and growth patterns that compared to those resulting from the hydrothermal carbonization by Fe 2+ ions under mild conditions, suggesting a common mechanism of formation. Textural changes of thin films found between [Zn][Al]Carbon spheres were attributed to differences in viscosity, which might have contributed to increases in functionality and specific surface area (by means of decreases in size) and, in turn, facilitating the sequestration of biomolecules. We propose that the presence of [Zn][Al]Carbon spheres provides protection against bacterial and UV attack. The physical properties of these spheres helped entrap biomolecules. Taken together, these factors contributed to the preservation of Pepita .

Patterns of gene flow vary greatly among Aedes aegypti populations throughout Mexico. The populations are panmictic along the Pacific coast, isolated by distance in northeast Mexico, and exhibit moderate gene flow across the Yucatan peninsula. Nine Ae. aegypti collections from 6 cities in Oaxaca, Mexico, were taken to examine the local patterns of gene flow. Genetic variation was examined in a 387-bp region of the nicotinamide adenine dinucleotide dehydrogenase subunit 4 mitochondrial gene (ND4) using single-strand conformation polymorphism analysis, and 3 haplotypes were detected. Cluster analysis on the linearized FST genetic distances failed to group collections in geographic proximity. Regression analysis of linear or road distances on linearized F(ST) indicated that proximal collections were as diverse as distant collections across an approximately 800-km range. The geographical distribution of the Mexican mosquito haplotype frequencies was determined for the ND4 sequences from 524 individuals from Oaxaca (this study) and 2,043 individuals from our previous studies. Herein, we report on yet another pattern dominated by genetic drift among 9 Ae. aegypti collections from 6 cities in Oaxaca, Mexico, and compare it to those reported in other regions of Mexico. Molecular analysis of variance showed that there was as much genetic variation among collections 4 km apart as there was among all collections. The numbers of haplotypes and the amount of genetic diversity among the collections from Oaxaca were much lower than detected in previous studies in other regions of Mexico and may reflect the effects of control efforts or adaptations to the altitudinal limits (1,500 m) of the species in Mexico. The geographical distribution of mosquito haplotypes in Mexico is also reported. Furthermore, based on the distribution of the mosquito haplotypes in America, we suggest that mosquito dispersion is very efficient, most likely due to commercial transportation.

Mitochondrial DNA mutations have been associated with different illnesses in humans, such as Kearns-Sayre syndrome (KSS), which is related to deletions of different sizes and positions among patients. Here, we report a Mexican patient with typical features of KSS containing a novel deletion of 7629 bp in size with 85% heteroplasmy, which has not been previously reported. Sequence analysis revealed 3-bp perfect short direct repeats flanking the deletion region, in addition to 7-bp imperfect direct repeats within 9-10 bp. Furthermore, sequencing, alignment and phylogenetic analysis of the hypervariable region revealed that the patient may belong to a founder Native American haplogroup C4c.

The main vector of dengue in America is the mosquitoAedes aegypti, which is infected by dengue virus (DENV) through receptors of midgut epithelial cells. The envelope protein (E) of dengue virus binds to receptors present on the host cells through its domain III that has been primarily recognized to bind cell receptors. In order to identify potential receptors, proteins from mosquito midgut tissue and C6/36 cells were purified by affinity using columns with the recombinant E protein domain III (rE-DIII) or DENV particles bound covalently to Sepharose 4B to compare and evaluate their performance to bind proteins including putative receptors from female mosquitoes ofAe. aegypti. To determine their identity mass spectrometric analysis of purified proteins separated by polyacrylamide gel electrophoresis was performed. Our results indicate that both viral particles and rE-DIII bound proteins with the same apparent molecular weights of 57 and 67 kDa. In addition, viral particles bound high molecular weight proteins. Purified proteins identified were enolase, beta-adrenergic receptor kinase (beta-ARK), translation elongation factor EF-1 alpha/Tu, and cadherin.

This study aims to portray the complex diversity of the Mexican Mestizo population, which represents 98.8% of the entire population of Mexico. We compiled extended haplotype data of the Y chromosome from populations in the Central Valley of Mexico (CVM), which we compared with other Mestizo and parental (Amerindian, European, and African) populations. A complex ancestral relationship was found in the CVM population, suggesting cosmopolitan origins. Nevertheless, the most preeminent lineages point toward a European ancestry, where the R1b lineage was most frequent. In addition, important frequencies of Amerindian lineages were also found in the Mestizo sample studied. Interestingly, the Amerindian ancestry showed a remarkable substructure, which was represented by the two main founding lineages: QL54 (× M3) and M3. However, even within each lineage a high diversity was found despite the small number of sample bearers of these lineages. Further, we detected important genetic differences between the CVM populations and the Mexican Mestizo populations from the north and south. This result points to the fact that Mestizo populations present different ancestral proportions, which are related to the demographic events that gave origin to each population. Finally, we provide additional forensic statistical parameters that are useful in the interpretation of genetic analysis where autosomal loci are limited. Our findings illustrate the complex genetic background of the Mexican Mestizo population and reinforce the need to encompass more geographic regions to generate more robust data for forensic applications.

Aedes aegypti from 24 collections in Mexico and the United States were challenged orally with dengue 2 virus JAM1409 (DEN-2 JAM1409). The vector competence (VC) of the populations ranged from 24% to 83%. Mosquito populations from the Yucatan exhibited greater VC than those from other areas of Mexico. The presence or absence of a midgut infection barrier (MIB) and a midgut escape barrier (MEB) was determined for mosquitoes in each population. The percentage of mosquitoes exhibiting an MIB ranged from 14% to 59%, and those exhibiting an MEB ranged from 4% to 43% in the collections. The MIB and MEB were not completely independent as determined by regression analysis. Midgut infection rates were dose dependent.

Aedes aegypti is the primary vector of yellow fever (YF) and dengue fever (DF) flaviviruses worldwide. In this review we focus on past and present research on genetic components and environmental factors in Aedes aegypti that appear to control flavivirus transmission. We review genetic relationships among Ae. aegypti populations throughout the world and discuss how variation in vector competence is correlated with overall genetic differences among populations. We describe current research into how genetic and environmental factors jointly affect distribution of vector competence in natural populations. Based on this information, we propose a population genetic model for vector competence and discuss our recent progress in testing this model. We end with a discussion of approaches being taken to identify the genes that may control flavivirus susceptibility in Ae. aegypti.

A population genetic analysis was conducted among 20 Aedes aegypti collections from 19 cities along the south Pacific coast in the Mexican states of Guerrero, Oaxaca, and Chiapas and in Coatepeque, Guatemala. Genetic variation was scored at 131 random amplified polymorphic DNA loci. The amount of genetic differentiation among collections was approximately 3 times as great as detected among collections in an earlier study in northeastern Mexico. Regression analysis of linear or road distances on linearized F(ST) indicated that collections are genetically isolated by distance. Cluster analysis failed to group collections in geographic proximity, and there was as much genetic variation among collections 60 km apart as there was among all collections (approximately 900-km range). The large genetic differentiation in southern Mexico reflects reduced gene flow among mosquitoes arising in a greater diversity of habitats and altitudes than exists among northeastern collections. It is likely that dispersal via human commerce in the northeast confounds patterns of natural gene flow.

A peculiar characteristic of Entamoeba histolytica trophozoites is their capacity to invade human tissues. One of the cellular determinants of invasion may include adhesion to extracellular matrix components such as collagen, induction, and secretion of electron-dense granules (EDG) and tissue digestion. The mechanism and receptors involved in this process are not well understood. Previous results suggested that cytoskeleton plays a very important role during EDG secretion. We present evidence suggesting that adhesion to collagen and activation of EDG secretion are integrin-dependent events, since β1 subunits detected by antibodies are concentrated at membrane sites where collagen and actin were colocalized. Furthermore, the involvement of actin, vimentin, and tubulin in restructuring cytoskeleton during EDG secretion was evident, since cytoskeleton isolation was possible exclusively in activated cells. Studies of immunolocalization of tubulin, actin, and vimentin by immunofluorescence and transmission electron microscopy suggest a role for cytoskeleton in EDG secretion. Cell Motil. Cytoskeleton 50:45–54, 2001. © 2001 Wiley-Liss, Inc.

A population genetic analysis of gene flow was conducted among 10 Aedes aegypti collections from seven cities along the northeastern coast of Mexico. Four collections were made from Monterrey to examine local patterns of gene flow. Markers included 60 random amplified polymorphic DNA (RAPD) loci amplified by the polymerase chain reaction and single strand conformation polymorphism analysis of variation in a 387-basepair region of the NADH dehydrogenase subunit 4 from the mitochondrial DNA (mtDNA). Seven mitochondrial haplotypes were detected and phylogenetic analysis identified two well-supported clades. Regression analysis of geographic distances and pairwise FST estimated from RAPD markers indicated that populations are isolated by distance and that free gene flow occurs among collections within 90-250 km. Isolation by distance was not detected using mtDNA haplotypes. The Nuevo Laredo collection had unique RAPD and mtDNA haplotype frequencies and reduced heterozygosity suggesting that few mosquitoes established this population.

The possible role of calmodulin (CaM) in functions such as cytotoxicity, phagocytosis, and amoebic growth was studied. These are important factors for the in vitro pathogenicity of Entamoeba histolytica HM 1 strain. The CaM inhibitors trifluoperazine (TFP), N-(6-aminohexyl)-5-chloro-1 -naphthalene-sulfonamide (W-7) and N-(6-aminohexyl)-naphthalenesulfonamide (W-5) were used for these purposes. Cytotoxicity was determined as the ability of amoebae to kill and/or lyse nucleated mouse spleen cells. Phagocytosis by amoebae was assessed by calculating the number of endocytosed human red blood cells. Cytotoxicity by E. histolytica showed a decrease dependent upon the concentration of W-7 and TFP, whereas W-5 did not show a significant inhibiting effect. Phagocytosis was roughly sensitive to W-7; TFP and W-5 did not have any significant effect. Amoebic growth was sensitive to TFP and W-7 CaM inhibitors. These results suggest that CaM participates in the expression of in vitro cytotoxicity of E. histolytica.

The protozoan Giardia lamblia initiates infection when trophozoites emerge from a cyst in the hosts by the excystation process. Although this process is crucial to the initiation of infection by G. lamblia, little is known about its regulation. To study the possible involvement of calmodulin (CaM) in excystation we tested the effect of several CaM antagonists (TFP, W-7, and W-5) on this cellular function. Except for W-5 the rest of these compounds inhibited excystation. The protein kinase C inhibitor H-7 had no effect on excystation, suggesting that CaM antagonists acted by selectively inhibiting CaM. Furthermore, CaM was redistributed after the induction of excystation and there was an increase in its fluorescence and activity. These results suggest that a CaM-dependent process is involved in G. lamblia excystation.

The interaction of Entamoeba histolytica with collagen induces the intracellular formation and release of electron-dense granules (EDGs) containing collagenase activity which are important in the pathogenicity of this parasite. Purified EDGs contain at least 25 polypeptides with acidic pIs, nine gelatinase activities, small molecules, including inorganic phosphate (Pi), pyrophosphate (PP) and other elements, including Na, Mg, S, Cl, K, Ca and Fe as measured by scanning transmission electron microscopy. Six of these polypeptides with apparent molecular weights of 108, 106, 104, 97, 68 and 59 kDa and two protease activities with apparent molecular weights of 40 and 85 kDa were detected exclusively in the EDGs and were not observed in total trophozoite extracts. Actin was also detected in the EDGs. Therefore, EDGs are a complex of mainly cationic proteins, which contains numerous proteolytic activities, actin and small molecules such as P(i), PP and cations.

Background Aedes aegypti is the primary global vector to humans of yellow fever and dengue flaviviruses. Over the past 50 years, many population genetic studies have documented large genetic differences among global populations of this species. These studies initially used morphological polymorphisms, followed later by allozymes, and most recently various molecular genetic markers including microsatellites and mitochondrial markers. In particular, since 2000, fourteen publications and four unpublished datasets have used sequence data from the NADH dehydrogenase subunit 4 mitochondrial gene to compare Ae. aegypti collections and collectively 95 unique mtDNA haplotypes have been found. Phylogenetic analyses in these many studies consistently resolved two clades but no comprehensive study of mtDNA haplotypes have been made in Africa, the continent in which the species originated. Methods and Findings ND4 haplotypes were sequenced in 426 Ae. aegypti s.l. from Senegal, West Africa and Kenya, East Africa. In Senegal 15 and in Kenya 7 new haplotypes were discovered. When added to the 95 published haplotypes and including 6 African Aedes species as outgroups, phylogenetic analyses showed that all but one Senegal haplotype occurred in a basal clade while most East African haplotypes occurred in a second clade arising from the basal clade. Globally distributed haplotypes occurred in both clades demonstrating that populations outside Africa consist of mixtures of mosquitoes from both clades. Conclusions Populations of Ae. aegypti outside Africa consist of mosquitoes arising from one of two ancestral clades. One clade is basal and primarily associated with West Africa while the second arises from the first and contains primarily mosquitoes from East Africa, Author Summary The authors are all medical entomologists who have worked in the field for more than 30 years. Over the past 20 years we have primarily worked on Aedes aegypti, the primary mosquito vector of Dengue and Yellow Fever Viruses. Twelve years ago, we began using mitochondrial markers to study relationships among Ae. aegypti populations. Since that time, 14 publications and 4 datasets have used the same markers and collectively 95 unique mtDNA haplotypes have been found. Haplotype phylogenies have consistently identified two clades. However, it wasn't until we combined efforts with our African colleagues that we realized that the two clades largely correspond with West and East Africa. Aedes aegypti populations from throughout the world are “mixtures” of mosquitoes from these two original clades. We plan to continue this effort to determine whether the composition of Ae. aegypti populations affects their ability to transmit arboviruses and also if mitochondrial haplotypes differ between mosquitoes with or without the newly discovered chromosomal inversions.

Glycine inhibits the formation of advanced glycation end products that may cause central and peripheral neuronal damage, affecting also the auditory nerve.To evaluate the effect of glycine on auditory nerve conduction and hearing level among patients with type 2 diabetes mellitus and auditory neuropathy.Twenty grams of oral glycine per day were administered during 6 months to 28 type 2 diabetic patients aged 58 ± 6 years, with auditory pathway neuropathy. Hearing tests and evoked otoacustic potentials were performed regularly. Fifteen diabetic patients aged 49 ± 8 years, without auditory nerve neuropathy did not receive glycine and were followed as a control group.Among patients receiving glycine, a significant improvement in left ear audiometry at 125, 250 and 500 Hz and right ear audiometry at 500 Hz, was observed. Waves I, III and V (p= 0.02) of evoked otoacustic potentials improved significantly in the left ear and wave I in the right ear. Among controls, waves V and III of evoked otoacoustic potentials had a significant impairment in the left ear.There was an improvement in auditory evoked potentials in patients receiving glycine and an impairment in untreated control patients.

Introduction : Glycine inhibits the formation of final products of advanced glication that occurs in hyperglycaemia causing central and peripherical neuronal damage affecting also the auditory nerve. Objective : Evaluate the effect of glycine on auditory nerve conduction and the hearing level in patients with diabetes mellitus type 2 and auditory neuropathy. Material and methods : Quasi-experimental study of pre-post test with control group nonequivalente design. Diabetic type 2 patients with neuropathy in the auditory pathway were included. Laboratory tests, hearing tests and evoked otoacoustic emissions were performed. Twenty grams of oral glycine per day were administered during 6 months. Results : In glycine group 28 patients were included and 15 in the control group, their average age was 57 and 48 years respectively. In the control group, left ear showed improvement at 125 Hz ( p = 0.02), 250 Hz ( p = 0.01) and 500 Hz ( p = 0.02), for right ear at 500 Hz ( p = 0.04). In the evoked otoacoustic emissions there was improvement in the left ear in the wave I ( p =0.01), III ( p =0.03) and V ( p = 0.02); in the right ear in the wave I ( p =0.03). The control group showed changes in the evoked otoacoustic emissions with significant damages in amplitud latent of the wave V (p=0.05) and in the wave III (p=0.03) for the left hear. Conclusions : G lycine partially improves nerve auditory conduction and hearing level, independently of the level of glicemic control.

1. 1. Calmodulin (CaM) was detected during secretion of electron-dense granules by Entamoeba histolytica trophozoites with immunofluorescence. 2. 2. It was purified to apparent homogeneity by chromatography with a yield of 2.26μg of calmodulin/mg of protozoan protein. Purity was established by gel electrophoresis. 3. 3. The parasite calmodulin has properties characteristic of calmodulin isolated from other eukaryotes: an apparent molecular weight of 19 or 17 kDa in presence of EGTA or CaCl 2 , respectively, activation in a calcium dependent manner of bovine heart cyclic nucleotide phosphodiesterase, and its UV spectrum.

Shoulder ultrasonography is revised in the evaluation of complete rotator cuff tears. Also another technique known as artro-ultrasonography, which consist of a 10 mL solution injection inside the articular capsule to observe its escape to the subacromial-subdeltoidea bursae which implicates rotator cuff tear. It is compared against multiple standards like surgery, magnetic resonance, arthrography and rehabilitation to include positive and negative cases.181 consecutive cases were studied with ultrasonography, 27 with artro-ultrasonography. The criteria for tear is the absence of the hyper-ecogenicity in the tendon, hypo-ecogenic gaps inside the rotator cuff and distention of the bursae due to the presence of liquid.69% sensibility and 71% specificity for ultrasonography. 92% sensibility and specificity for artro-ultrasonography. The area under the COR curve is 70% for ultrasonography and 93% for artro-ultrasonography. The artro-ultrasonography modified the diagnosis in nine cases, the majority from normal to tear. Three Radiologists proved the agreement with good results.The study contributes to evaluate the performance of ultrasound against multiple standards to correct the great variability in results observed in literature. The contribution of artro-ultrasonography to diagnosis is evaluated. It can be useful in places where accuracy is important and more sophisticated methods like magnetic resonance are not available.