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Vinculin is a protein found in both focal adhesions (FAs) and adherens junctions (AJs) which regulates actin connectivity to these structures. Many studies have demonstrated that mechanical perturbations of cells result in enhanced recruitment of vinculin to FAs and/or AJs. Likewise, many other studies have shown "cross-talk" between FAs and AJs. Vinculin itself has been suggested to be a probable regulator of this adhesion cross-talk. In this study we used MDCK as a model system of epithelia, developing cell lines in which vinculin recruitment was reduced or enhanced at AJs. Careful analysis of these cells revealed that perturbing vinculin recruitment to AJs resulted in a reduction of detectable FAs. Interestingly the cross-talk between these two structures was not due to a limited pool of vinculin, as increasing expression of vinculin did not rescue FA formation. Instead, we demonstrate that vinculin translocation between AJs and FAs is necessary for actin cytoskeleton rearrangements that occur during cell migration, which is necessary for large, well-formed FAs. Last, we show using a wound assay that collective cell migration is similarly hindered when vinculin recruitment is reduced or enhanced at AJs, highlighting that vinculin translocation between each compartment is necessary for efficient collective migration.

Bovine tuberculosis (TB) is a chronic disease caused mainly by Mycobacterium bovis, a zoonotic pathogen that has a worldwide distribution causing serious economic losses for milk and meat producers. In Chile, the disease in dairy cattle has a heterogeneous distribution, where the Metropolitan Region concentrates the highest animal prevalence and the main challenge for the national control and eradication programme. In this epidemiological context, vaccination with the M. bovis Bacillus Calmette-Guerin (BCG) vaccine might be a useful strategy for disease prevention and control. The objective of this study was to assess the efficacy and impacts on productivity and fertility of vaccination with the BCG Russia strain in 11 month-old heifers from a dairy farm, under a natural transmission condition. Sixty-two animals were vaccinated via the subcutaneous route with the equivalent of one human dose of BCG, and 60 control animals received saline. Subsequently, blood sampling was performed at 3, 6, 9, 12, 15 and 18 months post-inoculation, and infection status was determined using the IFNγ release assay (IGRA) with the DIVA (differentiate infected from vaccinated animals) antigens ESAT-6, CFP-10 and Rv3615c. Efficacy was calculated as the percentage of reduction in the incidence of infection attributable to vaccination, which showed a statistically significant level of overall protection of 66.5%. No adverse effects on fertility and production were recorded. In contrast, we observed beneficial effects of vaccination on several milk production parameters, with the milk yield in the first 100 days after calving in the BCG group significantly higher compared to unvaccinated heifers (p < .05). These results suggest that BCG vaccination of heifers in a natural transmission setting might result in both sanitary and productive benefits, supporting its implementation as a new strategy for TB prevention in a high prevalence area of Chile., (© 2021 Wiley-VCH GmbH.)

Bioinformatics analysis of the complete transcriptome of Fasciola hepatica, identified a total of ten putative carboxylesterase transcripts, including a 3146 bp mRNA transcript coding a 2205 bp open reading frame that translates into a protein of 735 amino acids, resulting in a predicted protein mass of 83.5 kDa and a putative carboxylesterase B enzyme. The gene coding for this enzyme was found in two reported F. hepatica complete genomes stretching 23,230 bp, containing two exons of 1282 and 1864 bp, respectively, as well as a 20,084 bp intron between the exons. The enzymatic activity was experimentally assayed on F. hepatica protein extracts by SDS-PAGE zymograms using synthetic chromogenic substrates, confirming both the theoretical molecular weight and carboxylesterase enzymatic activity. Further bioinformatics predicted that this enzyme is an integral component of the cellular membrane that should be active as a 167 kDa homodimer complex and polyacrylamide gel electrophoresis (PAGE) zymograms experiments confirmed the analysis. Additional bioinformatics analysis showed that DNA sequences that code for this particular enzyme are highly conserved in other parasitic trematodes, although they are labeled hypothetical proteins.

Genetic variants in the apolipoprotein E ( APOE ) gene are associated with lipid metabolism and lipid-related traits in the non-Hispanic population. There have been limited studies regarding the association between the APOE gene and hypercholesterolemia in the Hispanic population; therefore, our aim for this study is to examine the APOE gene's associations with cholesterol level and its related phenotypes. The APOE gene consists of three different alleles, ε2, ε3, and ε4, with ε4 being associated with dementia and cardiovascular diseases. A total of 1,382 subjects were collected from the Texas Alzheimer's Research and Care Consortium (TARCC, N = 1320) and the Initial Study of Longevity and Dementia from the Rio Grande Valley (ISLD-RGV, N = 62). Questionnaires on demographics, medical history, and blood/saliva samples were collected and APOE genotypes were performed. We observed allele frequencies of the APOE ε3 (96.7%), ε4 (22.6%) and ε2 (6.8%) alleles, respectively. Multivariable logistic regression revealed a significant association between the APOE ε4 allele and hypercholesteremia ( p = 1.8 × 10 -4 ) in our studied Hispanic population. We prove for the first time, that the APOE ε4 allele increases the risk for hypercholesterol in Hispanics. Further research is needed to confirm and supports our current findings.

It has been observed that there is a lower Parkinson's disease (PD) incidence in tobacco users. Nicotine is a cholinergic agonist and is the principal psychoactive compound in tobacco linked to cigarette addiction. Different studies have shown that nicotine has beneficial effects on sporadic and genetic models of PD. In this work we evaluate nicotine's protective effect in a Drosophila melanogaster model for PD where Synphilin-1 (Sph-1) is expressed in dopaminergic neurons. Nicotine has a moderate effect on dopaminergic neuron survival that becomes more evident as flies age. Nicotine is beneficial on fly survival and motility increasing tyrosine hydroxylase and dopamine levels, suggesting that cholinergic agonists may promote survival and metabolic function of the dopaminergic neurons that express Sph-1. The Sph-1 expressing fly is a good model for the study of early-onset phenotypes such as olfaction loss one of the main non-motor symptom related to PD. Our data suggest that nicotine is an interesting therapeutic molecule whose properties should be explored in future research on the phenotypic modulators of the disease and for the development of new treatments.

Fasciola hepatica is a worldwide distributed zoonotic parasitic trematode, which causes a severe liver disease clinically known as fasciolasis in a large number of wild animals, several livestock species as well as humans, prevention and control of fasciolasis is made by massive use of anthelmintic compounds on livestock and inevitably this practice has led to the emergence of anthelmintic resistant Fasciola hepatica and there is a great scientific effort to elucidate the molecular basis of anthelmintic resistance of parasitic helminths in general and of Fasciola hepatica in particular that may lead to improved anthelmintic compounds . In our project, we sequenced the transcriptomes obtained from the anthelmintic response to Triclabendazole and Albendazole on four samples from sensitive and resistant strains of Fasciola hepatica on Illumina HiSeq 4000 Platform and generated about 10.03 Gb per sample. The average genome-mapping rate is 81.29% and the average gene-mapping rate is 62.81%. 30,105 genes were identified in which 28,669 of them are known genes and 1,237 of them are novel genes from novel coding transcripts without any known features, 20,743 novel RNA transcripts were identified of which 14,293 of them are previously unknown splicing event for known genes but no alternative splicing was detected, the remaining 5,213 transcripts were found to be long noncoding RNA., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships, which have, or could be perceived to have, influenced the work reported in this article., (© 2021 The Author(s).)

Liberibacter is a bacterial group causing different diseases and disorders in plants. Among liberibacters, Candidatus Liberibacter solanaceraum (CLso) produces disorders in several species mainly within Apiaceae and Solanaceae families. CLso isolates are usually grouped in defined haplotypes according to single nucleotide polymorphisms in genes associated with ribosomal elements. In order to characterize more precisely isolates of CLso identified in potato in Spain, a Multilocus Sequence Analysis (MLSA) was applied. This methodology was validated by a complete analysis of ten housekeeping genes that showed an absence of positive selection and a nearly neutral mechanism for their evolution. Most of the analysis performed with single housekeeping genes, as well as MLSA, grouped together isolates of CLso detected in potato crops in Spain within the haplotype E, undistinguishable from those infecting carrots, parsnips or celery. Moreover, the information from these housekeeping genes was used to estimate the evolutionary divergence among the different CLso by using the concatenated sequences of the genes assayed. Data obtained on the divergence among CLso haplotypes support the hypothesis of evolutionary events connected with different hosts, in different geographic areas, and possibly associated with different vectors. Our results demonstrate the absence in Spain of CLso isolates molecularly classified as haplotypes A and B, traditionally considered causal agents of zebra chip in potato, as well as the uncertain possibility of the present haplotype to produce major disease outbreaks in potato that may depend on many factors that should be further evaluated in future works.

Background: Cervical Cancer (CC) is a worldwide public health concern associated with genetic alterations, among these the gain of the 19q chromosome harboring the Pregnancy Specific Glycoproteins (PSG) gene family. These proteins play a critical role in pregnancy, with participation in immunotolerance, angiogenesis, and invasion processes, which are also observed in carcinogenesis. The aim of this study was to determine the molecular alterations of PSG1 and its relationship with CC., Methods: PSG1 Copy Number Variation (CNV) was evaluated in 31 CC and eight normal cervical tissues by qPCR. PSG1 expression was correlated with HPV detection and IL-10 and TGF-β expression in CC samples. Finally, PSG1 protein expression was evaluated by immunofluorescence in CC cell lines, by immunohistochemistry in a tissue microarray, and by immunoblotting in the sera of women with normal cervix, pre-invasive lesions, and CC., Results: PSG1 showed a gain of 25.6% in CNV and gene expression in CC. There was a lack of PSG1 expression in normal cervical epithelium and positive immunostaining in 57% of CC tissues, while all CC cell lines expressed PSG1. Finally, PSG1 was immunodetected in 90% of pre-invasive lesions and in all CC serum samples, but not in healthy women. PSG1 expression correlates with the expression of IL-10 and TGF-β in CC tissues, but not with the presence of HPV., Conclusion: These data show evidence of the differential expression of PSG1 in CC that could explain its participation in tumor-biology and immunotolerance mechanisms. Further, its immunodetection could provide early detection of this cancer., (Copyright © 2020 IMSS. Published by Elsevier Inc. All rights reserved.)

Background: Acaricide resistance is a central problem for the control of the cattle tick Rhipicephalus microplus. The physiological effects and phenotypes of the mutations that cause acaricide resistance are not always well understood or characterized. Single nucleotide polymorphisms (SNPs) that confer cypermethrin knockdown resistance (kdr) have been reported in R. microplus. These SNPs have been associated and correlated with pyrethroid resistance although there is no direct physiological evidence that their presence does confer kdr in this organism., Methods: Resistant and susceptible strain resistance profiles were obtained using the larval packet discriminating dose assay. The relevant genomic regions of the para-sodium channel were amplified using standard PCR; SNPs were detected by sequencing the corresponding amplicons. Ovary response to cypermethrin exposure/treatment was evaluated using videometrical analysis., Results: We found that the pyrethroid resistance trait is stable in a resistant reference strain after years without selection, suggesting that the resistance conferring mutations are fixed in the population. In this strain, a change in the structure of the pre-synaptic para-sodium channel caused by the G184C, the C190A and the T2134A SNPs appears to confer resistance. These mutations are absent in the susceptible strain used as control. We demonstrate that cypermethrin blocks ovary contraction in cypermethrin-susceptible ticks. We also show that ovaries from organisms that carry the kdr associated SNPs still contract at cypermethrin concentrations that completely block ovary contraction in the susceptible strain. The configuration of the experimental system excludes a xenobiotic detoxification mechanism., Conclusions: This is the first report that presents physiological evidence that the presence of the G184C, the C190A, and the T2134A mutations in the para-sodium channel correlates with maintaining muscle contractility in R. microplus exposed to cypermethrin. These SNPs may confer cypermethrin resistance in this organism by avoiding presynaptic blockage, inhibiting the flaccid muscle paralysis characteristic of this acaricide. The videometric assay that we previously validated can be used to detect more rapidly than other assays that involve larval mortality kdr-like cypermethrin resistant tick strains, permitting to directly assay adult pre-engorged females after they are collected on the field without waiting until eggs are laid and larvae eclose.

The dysfunction of the proteasome-ubiquitin system is commonly reported in several neurodegenerative diseases. Post mortem samples of brains of patients with Parkinson´s disease present cytoplasmic inclusions that are rich in proteins such as ubiquitin, Tau, and α-synuclein. In Parkinson´s disease, a specific reduction of some of the proteasome subunits has also been reported. However, the specific role of the different proteasome subunits in dopaminergic neuron degeneration has not been thoroughly explored. In this work, we used the Gal4/UAS system to test fourteen Drosophila melanogaster RNAi lines from the Bloomington Drosophila Stock Center. Each of these lines targets a different proteasome subunit. To identify the strains that were able to induce neurodegeneration, we drove the expression of these lines to the eye and cataloged them as a function of the extent of neurodegeneration that they induced. The targeted proteasomal subunits are conserved in mammals and therefore may be relevant to study proteasome related diseases. The RNAi line among the regulatory subunits with the most penetrant phenotype targeted the proteasomal subunit Rpt2 and we decided to further characterize its phenotypes. Rpt2 knockdown in the Drosophila central nervous system reduced the activity of the proteasome, augmented the amount of insoluble ubiquitinated protein, and elicited motor and non-motor phenotypes that were similar to the ones found in Drosophila and other models for Parkinson's disease. When Rpt2 is silenced pan-neurally, third instar larvae have locomotion dysfunctions and die during pupation. Larval lethality was avoided using the Gal80-Gal4 system to induce the expression of the Rpt2 RNAi to dopaminergic neurons only after pupation. The reduction of Rpt2 in adult dopaminergic neurons causes reduced survival, hyperactivity, neurodegeneration, and sleep loss; probably recapitulating some of the sleep disorders that Parkinson's disease patients have before the appearance of locomotion disorders., Competing Interests: We also declare no conflict of interests., (© 2020 The Author(s).)

Isolates of the Tomato yellow leaf curl virus (TYLCV) species (genus Begomovirus , family Geminiviridae ) infect tomato crops worldwide, causing severe economic damage. Members of the whitefly Bemisia tabaci sibling species group are the vector of begomoviruses, including TYLCV. However, transmission of isolates of the type strain (Israel [IL]) of TYLCV (TYLCV-IL) by tomato seed has recently been reported based on infections occurring in Korea. Because of the consequences of this finding on the epidemiology and control of the disease caused by TYLCV and on the seed market, it was considered essential to revisit and expand those results to other tomato-growing areas. TYLCV DNA content was detected in tomato and Nicotiana benthamiana seed collected from plants naturally or experimentally infected with TYLCV-IL, supporting its seedborne nature. The TYLCV-IL replication detected in tomato and N. benthamiana flower reproductive organs demonstrated close association of this virus with the seed during maturation. However, the significant reduction of TYLCV DNA load after surface disinfections of tomato seed suggests that most of the virus is located externally, as contaminant of the seed coat. Transmission assays, carried out with seven tomato genotypes and more than 3,000 tomato plants, revealed no evidence of seed transmission from "surface-disinfected" or untreated seed for two Mediterranean isolates of TYLCV-IL. Similar results were also obtained for seed collected from TYLCV-IL-infected N. benthamiana plants. The results support the conclusion that TYLCV-IL is seedborne but is not seed transmitted in tomato or N. benthamiana , suggesting that transmission through seed is not a general property of TYLCV.

In the present study, titanium (IV) sulfide (TiS 2 ) was synthesized and investigated for the removal of Cu 2+ and Pb 2+ ions from aqueous solutions. TiS 2 nanoparticles synthesized through a solvothermal synthesis were characterized using x-ray diffraction (XRD) and scanning electron microscopy (SEM). The average particle size for the TiS 2 material was determined to be 8.03 ± 0.98 nm from the diffraction pattern. Studies were performed to examine the effects of pH, temperature, time, and interfering ions on the binding of Cu 2+ and Pb 2+ to the TiS 2 . As well isotherm studies were performed to determine the binding capacity of TiS 2 for both Cu 2+ and Pb 2+ ions. The pH profile studies showed optimal binding occurred at pH 2 for the sorption of both Cu 2+ and Pb 2+ to the TiS 2 . The isotherm studies showed the adsorption capacities at temperatures of 4, 22, and 45°C for Cu 2+ were 243, 222, and 153 mg/g, respectively. An opposite trend in the adsorption was observed for Pb 2+ binding to the TiS 2 . The observed binding capacities for Pb 2+ were 32, 166, and 357 mg/g, at temperatures of 4, 22, and 45°C, respectively. The thermodynamic parameters for binding showed a non-spontaneous process for the sorption of Cu 2+ whereas a spontaneous binding process was observed for the sorption of Pb 2+ . Additionally, the binding of Cu 2+ on TiS 2 in the presence of interfering ions (Na + , K + , Mg 2+ , and/or Ca 2+ ) was observed to decrease at high concentrations; however, the binding of Pb 2+ was unaffected by the presence of the same cations.

Aims: Gynaecological malignancies such as breast, ovarian and cervical cancers have become an important public health problem. Detection of molecular alterations in cancer research is fundamental since it can reveal specific pathogenic patterns and genes that could serve as markers. Our aim was to characterize common genomic and transcriptomic signatures for the three gynaecologic malignancies with the highest incidence and mortality to try to identify new molecular markers, therapeutic targets and molecular signatures., Methods: Here we analysed a total of 723 microarray libraries corresponding to equal number of breast, ovary and cervical cancer and non-cancer patient samples. Copy number variation (CNV) was carried out using 428 libraries and transcriptomic analysis using the 295 remaining samples., Results: Our results showed that breast, ovary and cervical malignancies are characterized by gain of 1q chromosome. At transcriptomic level, they share 351 coding and non-coding genes, which could represent core transcriptome of gynaecological malignancies. Pathway analysis from the resulting gene lists from CNV and expression showed participation in cell cycle, metabolism, and cell adhesion molecules among others., Conclusions: Chromosome 1q characterize the gynaecological malignancies, which could harbour a richness of genetic repertoire to mine for molecular markers and targets, particular gynaecologic expression profile, containing FANCI, FH and MIR155HG among others, could represent part of the transcriptomic core for diagnostic test and attractive therapeutic targets. It may not be long before every human cancer sample is profiled for a detections test to ascertain a molecular diagnosis and prognosis and to define an optimal and precise treatment strategy., (Copyright © 2019 IMSS. Published by Elsevier Inc. All rights reserved.)

Cervical cancer (CC) is associated with alterations in immune system balance, which is primarily due to a shift from Th1 to Th2 and the unbalance of Th17/Treg cells. Using in silico DNA copy number analysis, we have demonstrated that ~20% of CC samples exhibit gain of 8q22.3 and 19q13.31; the regions of the genome that encodes the KLF10 and PSG genes, respectively. Gene expression studies demonstrated that there were no alterations in KLF10 mRNA expression, whilst the PSG2 and -5 genes were up-regulated by 1.76 and 3.97-fold respectively in CC compared to normal tissue controls. siRNA and ChIP experiments in SiHa cells have demonstrated that KLF10 participates in immune response through regulation of IL6, IL25 and PSG2 and PSG5 genes. Using cervical tissues from KLF10 -/- mice, we have identified down-regulation of PSG17, -21 and -23 and IL11. These results suggest that KLF10 may regulate immune system response genes in cervical cancer among other functions. KLF10 and PSG copy number variations and alterations in mRNA expression levels could represent novel molecular markers in CC.

Background: Magnesium (Mg) deficiency contributes to the pathophysiology of numerous diseases. The therapeutic use of Mg has steadily increased over time. The increased in-hospital use of intravenous (IV) magnesium sulfate (MgSO4) warrants more extensive investigation regarding the safety of the therapy. The aim of this study was to determine the safety of IV MgSO4 infusion on cardiovascular, liver, kidney, and metabolic markers in adults., Methods: Twelve volunteers were randomized to one of two cross-over conditions: (a) IV infusion of MgSO4 in 5% dextrose followed by IV infusion of 5% dextrose 1 week later or (b) IV infusion of 5% dextrose followed by IV infusion of MgSO4 in 5% dextrose 1 week later. An electrocardiogram was recorded continuously during the infusions. Blood was drawn pre- and post-infusion for blood count (high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and triglycerides). Results: Serum Mg increased from pre- to post-infusion in the MgSO4 + 5% dextrose group (p < 0.0001). The QRS interval length increased from pre- to post-infusion in the MgSO4 + 5% dextrose group (p < 0.04). Additionally, serum glucose concentration increased in the MgSO4 + 5% dextrose group (p = 0.04). These significant findings were modeled with gender and age as covariates. No other significant differences were found., Conclusions: The administration of IV infusion of MgSO4 (4 g/100 mL) in 5% dextrose over a 4-hour treatment period poses no significant deleterious effects on cardiovascular, liver, kidney, or metabolic function., Relevance for Patients: IV infusion of MgSO4 may be used for certain treatment indications without significant concern for systemic or organ toxicity.

The effects of the immune system response in the malignant transformation process have been described. Molecules such as interferons are involved in such process. Interferons are small single-chained glycoproteins, involved in the first line of defense against pathogens such as viruses, bacteria, and parasites. Interferon epsilon (IFNε) is located in the 9p21.3 cytogenetic region, transcribes into a single exon mRNA. Contrary to other family members, IFNε exerts low antiviral activity. In the present work molecular alterations such as copy number variation (CNV) and expression were analyzed by available microarrays and fifty-nine cervical tissues ranging from normal to cancer and three cell lines were assessed for IFNε expression by RT-PCR, immunohistochemistry, and immunocytofluorescence. No significant CNV alterations were observed. Positive immunosignal was primarily present in the proliferative basal strata cells in the normal tissue, whereas in cervical cancer, all epithelial transformed cells were positive. The cell lines analyzed were HPV16, -18, and negative, all three cell-lines were positive for cytoplasmic protein presence. Interestingly, at the mRNA level, increased band intensity was observed, as the lesions were higher, and IFNε up-regulation in CC ( P=0.0001 ) is reported here. Our results suggest that up-regulation is present as an independent event from single or multiple HPV infection ( P=0.90 ). In conclusion, we suggest that IFNε mRNA up-regulation could represent a potential molecular marker in CC. Expression of IFNε might not be related to HPV infection or CNV, which could have an important role in cellular homeostasis and could influence immune related events in cervical carcinogenesis., Competing Interests: None., (IJCEP Copyright © 2018.)

Background: Similarities between the pathologic progression of cancer and the physiologic process of placentation have been recognized for many years proposing that both present similar mechanisms and processes. Cervical cancer (CC) is one of the most frequent neoplasia among Mexican women turning it into an important health problem., Objective: The aim of this study was to determine the degree of the involvement of pregnancy related genes and in cancer progression by in-silico analysis and validated in CC samples., Results: The data mining analysis resulted in the identification of genes expressed in term placenta, first trimester placenta and normal cervical tissues. Finally, we selected KISS1 for the involvement of pregnancy related gene and also in cancer process. In order to explore KISS1 in CC, we analyzed Copy Number Variation (CNV) and gene expression using microarray experiments. KISS1 showed 20% genomic gain in 1q32.1 on CC samples. Furthermore, microarray analysis showed KISS1 as up-regulated genes. Results were validated showing an overexpression of 85% of KISS1 in CC samples., Conclusions: Data suggest KISS1 as a great candidate for CC molecular markers or as a therapeutic target for CC. Also, HPV presence does not seem to alter the KISS1 expression in CC.

Background and Aims: Krüppel Like Factors (KLF) refers to a family of seventeen members of transcription factors. Involved in several cellular processes. As other cancer types, Cervical Cancer (CC) presents molecular deregulations in transcription factors, but especially Human Papilloma Virus (HPV) sequences. Here in this work we analyzed the mRNA expression of all KLF family members in CC-derived cell lines and CC tissues., Methods: The cell lines used were HeLa, INBL, RoVa, C4-I, Ms751, ViPa, CaLo, SiHa, CaSki, C33a and ViBo and the non-tumorigenic HaCaT. mRNA expression was analyzed by means of expression microarray and RT-PCR, and KLF5 protein by immunofluorescence., Results: The cell lines were grouped according to HPV genotype as HPV16, HPV18 positive or HPV negative cells. Heterogeneous expression was observed among the cell lines. Despite the heterogeneous expression profile, KLF3, -5, -12, -15 and -16 transcripts were present in all cell lines, KLF4 and -10 which were not expressed in CaSki; KLF11 and 13 were not expressed by Vipa and C4-I, and KLF7 was not expressed by C4-I and Rova. The CC tissue analysis shows expression of most of the KLF members, such as KLF5. KLF5 immunosignal was positive in the three cell lines analyzed., Conclusions: We suggest that KLF expression could not be related to HPV presence/genotype, at least at transcriptional level, and the expression of KLF family members may be necessary in the biology of the CC cells., (Copyright © 2017 IMSS. Published by Elsevier Inc. All rights reserved.)

Aim: Treatment-resistant depression patients are more likely to suffer from comorbid physical and mental disorders, experience marked and protracted functional impairment, and incur higher health-care costs than non-affected individuals. Magnesium sulfate is a treatment option that may offer great potential for patients with treatment-resistant depression based on prior work in animals and humans., Methods: Twelve subjects with mild or moderate treatment-resistant depression were randomized into a double-blind crossover trial to receive an infusion of 4 g of magnesium sulfate in 5% dextrose or placebo infusion of 5% dextrose with a 5-day washout in between the 8-day intervention period. Subjects were assessed before and after the intervention for serum and urine magnesium, lipid panel, the Hamilton Rating Scale for Depression, and the Patient Health Questionnaire-9., Results: We found a difference in serum magnesium from day 2 to 8 (pre-infusion) (P < 0.002) and from baseline to day 8 (P < 0.02). No changes were noted on the Hamilton Rating Scale for Depression or the Patient Health Questionnaire-9 24 h post-treatment, but as serum magnesium increased from baseline to day 7, the Patient Health Questionnaire-9 decreased from baseline to day 7 (P = 0.02)., Conclusion: Magnesium sulfate did not significantly affect depression 24 h post-infusion, but other results were consistent with the literature. The association between changes in serum magnesium and the Patient Health Questionnaire-9 supports the idea that magnesium sulfate may be used to address treatment-resistant depression, an ongoing medical challenge., (© 2016 The Authors Psychiatry and Clinical Neurosciences © 2016 Japanese Society of Psychiatry and Neurology.)

Background: Cervical Cancer (CC) has become a public health concern of alarming proportions in many developing countries such as Mexico, particularly in low income sectors and marginalized regions. As such, an early detection is a key medical factor in improving not only their population's quality of life but also its life expectancy. Interestingly, there has been an increase in the number of reports describing successful attempts at detecting cancer cells in human tissues or fluids using trained (sniffer) dogs. The great odor detection threshold exhibited by dogs is not unheard of. However, this represented a potential opportunity to develop an affordable, accessible, and non-invasive method for detection of CC., Methods: Using clicker training, a male beagle was trained to recognize CC odor. During training, fresh CC biopsies were used as a reference point. Other samples used included cervical smears on glass slides and medical surgical bandages used as intimate sanitary pads by CC patients. A double-blind procedure was exercised when testing the beagle's ability to discriminate CC from control samples., Results: The beagle was proven able to detect CC-specific volatile organic compounds (VOC) contained in both fresh cervical smear samples and adsorbent material samples. Beagle's success rate at detecting and discriminating CC and non-CC odors, as indicated by specificity and sensitivity values recorded during the experiment, stood at an overall high (>90%). CC-related VOC in adsorbent materials were detectable after only eight hours of use by CC patients., Conclusion: Present data suggests different applications for VOC from the uterine cervix to be used in the detection and diagnosis of CC. Furthermore, data supports the use of trained dogs as a viable, affordable, non-invasive and, therefore, highly relevant alternative method for detection of CC lesions. Additional benefits of this method include its quick turnaround time and ease of use while remaining highly accurate and robust.

There are more people in the world in the youth stage of development than in any other stage of development. The youth in Puerto Rico faces many situations that affect their development and readiness for adulthood. Therefore, it's imperative to identify evidence-based positive youth development interventions to develop practices that help young people prevent adverse situations, promote positive experiences and encourage children and young people to be involved and committed. One hundred and forty seven interventions were identified through a traditional scientific literature review. The results reflect that the interventions mainly focused on reducing risk factors and increasing of protection factors. However, no intervention focus on fully engaging children and young people to be involved and committed to their optimal development and to their communities. Nevertheless, all identified interventions providDe tools that could be useful to foster such practices in the context of Puerto Rico. Of the 147 interventions identified, six are designed for the Puerto Rican population residing on the island. In order to make information accessible to professionals and the community, the collection of the Archivo de Programas y Prácticas Basadas en Evidencia para la Prevención has been expanded.

The tyraminergic/octopaminergic system is central for the control of arthropod oviposition. Previous works demonstrated that the pharmacological perturbation of this system inhibits oviposition in the cattle tick Rhipicephalus microplus. In this work, we describe a physiologically active whole-mount preparation of the contractile tick ovary that allows the quantitative videometrical analysis of ovary contraction in response to different compounds. Eight adrenergic ligands known to inhibit oviposition, including octopamine and tyramine were tested. These compounds exhibited antagonistic effects; octopamine relaxes the ovary preparation while tyramine induces a very strong contraction. The other adrenergic compounds tested were classified as able to contract or relax ovary muscle tissue. Isoprotenerol has a stronger relaxative effect than octopamine. Tyramine induces the biggest contraction observed of all the compounds tested, followed, in descending amount of contraction, by salbutamol, prazosin, epinastine, clonidine and the acaricide amitraz. The effect of these adrenergic ligands on the ovary preparation, explains why these molecules inhibit tick oviposition and suggest a regulatory mechanism for ovary contraction and relaxation during oviposition. Our results also provide a physiological explanation of the egg-laying inhibition effect of amitraz when used on the cattle tick.

Thymopoietin (TMPO) is an inner nuclear membrane protein, the coding gene named equally, can give arise to six isoforms by alternative splicing. This gene has been found up regulated in several types of cancer. At present work, we evaluated the TMPO isoforms generated by alternative splicing as well as the protein signal detection in breast cancer samples. TMPO expression was analyzed by immunohistochemistry in tissue microarray containing 46 breast tissue samples including normal (n = 6), benign lesions (n = 18) (fibroadenomas (n = 6), fibrocystic changes (n = 6), ductal hyperplasias (n = 6)) and breast carcinoma (n = 22). Isoforms -α, -β and -γ of TMPO were evaluated using RT-PCR; clinical-pathological correlation analysis were done by mean of X(2). Neither the normal nor the benign lesions of the breast showed positive TMPO immunodetection, whilst 45 % of the breast carcinomas were immunopositive (p = 0.000), nine of ten positives carcinomas correspond to the Luminal A subtype. Further, alpha isoform was present in all breast samples analyzed; however, beta and gamma isoforms were only present in ten (p = 0.003) and 17 (p = 0.000), respectively, in the breast cancer samples. According with the present data, we suggest that TMPOβ and -γ isoforms could provide a potential reliable diagnostic marker for breast cancer.

In humans, there is a strong correlation between sensitivity to substances of abuse and addiction risk. This differential tolerance to drugs has a strong genetic component. The identification of human genetic factors that alter drug tolerance has been a difficult task. For this reason and taking advantage of the fact that Drosophila responds similarly to humans to many drugs, and that genetically it has a high degree of homology (sharing at least 70% of genes known to be involved in human genetic diseases), we looked for genes in Drosophila that altered their nicotine sensitivity. We developed an instantaneous nicotine vaporization technique that exposed flies in a reproducible way. The amount of nicotine sufficient to "knock out" half of control flies for 30 minutes was determined and this parameter was defined as Half Recovery Time (HRT). Two fly lines, L4 and L70, whose HRT was significantly longer than control´s were identified. The L4 insertion is a loss of function allele of the transcriptional factor escargot (esg), whereas L70 insertion causes miss-expression of the microRNA cluster miR-310-311-312-313 (miR-310c). In this work, we demonstrate that esg loss of function induces nicotine sensitivity possibly by altering development of sensory organs and neurons in the medial section of the thoracoabdominal ganglion. The ectopic expression of the miR-310c also induces nicotine sensitivity by lowering Esg levels thus disrupting sensory organs and possibly to the modulation of other miR-310c targets.

Unlabelled: The intra-abdominal umbilical vein varix is very uncommon, characterized by a dilation of the vein, usually located between its entry into the abdomen and completion in the portal system. However, the location of this anomaly to extra-abdominal level is very unusual and more difficult to diagnose by ultrasound., Clinical Case: We present a case undiagnosed during pregnancy, demonstrating cord abnormality in its extra-abdominal portion and confirmed later by the pathology study. It was a gestation of 36 + 6 weeks, ending by performing emergency caesarean section for abnormal fetal heart rate, with good perinatal outcome. The diagnosis was made after placental delivery, by the existence of fibrotic cystic structure in the middle third of the umbilical cord.

Cervical cancer (CC) is a multifactorial disease associated to genetic, environmental and epigenetic factors, being the infection by human papillomavirus the main etiologic agent. Additionally, the alteration in the expression of transcription factors has been considered of importance for the development of this tumor. HOX genes encode a group of transcription factors involved in cellular proliferation and differentiation processes during the development of embryonic structures in vertebrates; their aberrant expression is associated with tumorigenesis and metastasis. A range of evidence suggests a role for HOX genes in the development of cervical neoplastic cell. Studies in CC cell lines, primary tumors and premalignant lesions have suggested the involvement of HOXA1, HOXC5, C6, C8 and C10, HOXD9 and HOXD13 in the process of cervical carcinogenesis. Also, the de novo expression of genes HOXB2, B4, B13 and HOXC11-C13 appears to be involved in the process of malignant transformation of cervical epithelial cell. These data would allow to open a field in search of new molecular markers in cervical cancer and the development of new therapeutic strategies for this malignancy.

Chagas disease, or American trypanosomiasis, which is caused by the protozoan parasite Trypanosoma cruzi, is primarily a vector disease endemic in 21 Latin American countries, including Mexico. Although many vector control programs have been implemented, T. cruzi has not been eradicated. The development of an anti-T. cruzi vaccine for prophylactic and therapeutic purposes may significantly contribute to the transmission control of Chagas disease. Immune protection against experimental infection with T. cruzi has been studied since the second decade of the last century, and many types of immunogens have been used subsequently, such as killed or attenuated parasites and new DNA vaccines. This primary prevention strategy appears feasible, effective, safe, and inexpensive, although problems remain. The objective of this review is to summarize the research efforts about the development of vaccines against Chagas disease worldwide. A thorough literature review was conducted by searching PubMed with the terms "Chagas disease" and "American trypanosomiasis" together with "vaccines" or "immunization". In addition, reports and journals not cited in PubMed were identified. Publications in English, Spanish, and Portuguese were reviewed.

Dogs and coyotes are the most frequently reported canids responsible for scavenging human remains. We present the case of a 90-year-old woman whose mummified body was found in her home showing partial destruction of the thorax and extremities and absence of the cranium. The victim lived with a beagle dog whose dead body was also found, along with abundant scats throughout the house. Scavenging by the decedent's pet was the proposed hypothesis for the partial dismemberment and consumption of her body. Forensic analysis revealed that the victim died as a result of an accidental fracture of the proximal femoral epiphysis. Bone exam showed signs of canine scavenging on certain bones. Macroscopic and histological analyses of the dog feces revealed the presence of small bone fragments within scats. All the collected data supported the hypothesis that the decedent's pet fed on the victim following her death. The current case illustrates that forensic anthropology has much more to offer than personal identification and determining the manner of death. Systematic search and examination of scat deposits recovered from the scene may be very useful in the medicolegal investigation, identifying the origin of body mutilation and particularly the animal responsible for any scavenging., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The cattle tick Rhipicephalus microplus, is one of the most damaging livestock ectoparasites. Tropical tick infestation limits the introduction of high-yield bovine varieties because they do not have immunity to the diseases transmitted by these ectoparasites. This tick is usually controlled with chemical acaricides but their indiscriminate use has created resistant populations. The discovery of new molecules that can be used for tick control is urgent. Based on the knowledge that octopamine, a biogenic amine analog to epinephrine, is central to the regulation of oviposition in several studied arthropods and that an imbalance in octopamine release causes sterility in a Drosophila model. Tyramine, octopamine and epinastine and 83 adrenergic compounds classified by their effect in the vertebrate systems were screened for their ability to block oviposition in Rhipicephalus microplus. Of these molecules, we found that 10 alpha-agonists, 3 alpha-antagonists, 5 beta-adrenergic agonists, 7 beta-antagonists and Norepinephrine were able to inhibit oviposition in this tick at pharmacological concentrations. Surprisingly, tyramine appears to be more potent than octopamine. The probable physiological causes of this inhibition are discussed. Our results suggest that although there are alpha adrenergic-like receptors in the tick, they do not behave in a manner completely analogous to their vertebrate counterparts., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Purpose: The aim of this study is to investigate the relationship between invasive brain tissue oxygen pressure (PbrO(2)) and noninvasive regional transcranial oxygen saturation (rSO(2)) in 22 stable patients with severe traumatic brain injury (TBI) during a 16 h period., Methods: This was a prospective, observational study carried out in the Neurocritical Care Unit of a level 1 trauma center in a teaching hospital. A total of 41,809 paired records for neuromonitoring variables were analyzed and compared., Results: A direct and independent correlation between rSO(2) and PbrO(2) was confirmed through adjusted [beta coefficient and (95% confidence interval, CI) = 0.36 (0.35-0.37)] and logistic [PbrO(2) >or=15 mmHg, as a dependent variable; adjusted odds ratio (AOR) and (95% CI) = 1.11 (1.10-1.12)] regression analyses. A receiver-operating characteristic (ROC) curve demonstrated that rSO(2) had low accuracy for detecting moderate (PbrO(2)

Background: Neuroblastoma (NB) is the principal tumor of the sympathetic nervous system in children., Objective: to describe the epidemiological and clinical characteristics of Mexican children with NB., Methods: A population-based, prolective study, with data obtained from the Childhood Cancer Registry of the Instituto Mexicano del Seguro Social. The frequencies and incidence of the variables of the study were obtained by age and sex. The trend was calculated with the annual percentage of change., Results: Of a total of 2758 children with cancer, 72 (2.6%) were identified in the Group IV, according to the International Classification of Childhood Cancer. The average incidence for NB was 3.8, the highest incidence was found in Guerrero. NB was highest in the group under one year of age (18.5). The male/female ratio was 1.1 and there was no trend toward an increase. Stages III and IV were presented in 88% of the cases. There was no association between the stages, the age at the TD, or the histological pattern., Conclusions: It is possible that the low incidence of NB in Mexican children is due to the difficulty in early diagnosis due the majority of the cases was diagnosed in the advanced stages.

Objective: to present the frequency of different types of cancer in adolescents with 15 and 16-year-old treated at the Instituto Mexicano del Seguro Social (IMSS) in Mexico City (MC) hospitals., Methods: it was an observational and prolective study. During January 1 of 1996 to December 31 of 2005, were analyzed new cases of adolescents of 15 and 16-year-old with malignant neoplasms in adolescents at IMSS. The neo-plasms were grouped according with the International Classification of Childhood Cancer; they were stratified by age, sex, and place of residence, and the frequency of these variables were obtained., Results: a total of 207 cases were registered. Leukemias were more frequent with 55 cases (26.6%); the second place was for bone tumors with 36 cases (17.4%) and the third place were lymphomas with 32 cases (15.5%). There were not cases of retinoblastoma and renal tumors; only 8 cases of carcinomas (3.9%) were registered. The greatest frequency was observed in males with 116 cases (56.0%), with male/female ratio of 1.3:1. Mexico City and State of Mexico were more frequent places of residence with 70.5% of cases. Near of forty nine percent of cases (48.8%) with solid tumors were diagnosed in III or IV stages., Conclusions: the pattern of cancer was different from older adolescents (15 to 19-year-old), it is necessary to conduct studies of survey in order to determine epidemiology of cancer in the Mexican adolescents.

Objective: To determine the epidemiology of cancer among children from eleven Mexican states affiliated to Mexican Social Security Institute (Instituto Mexicano del Seguro Social, IMSS)., Methods: 2663 Cases of children with cancer were prospectively registered between 1996 and 2002 in the main IMSS medical centers. The cases were classified according to the International Classification for Childhood Cancer. The general and specific cancer incidences (rate per million children/year, age standardized) were obtained., Results: The principal groups of cancer in nine states were leukemias, lymphomas and central nervous system tumors (CNST). In eight states, the incidence was similar than reported worldwide (100 to 180); Chiapas had the highest incidence (203.5). Leukemias had higher incidence in seven States (>50). The highest incidences of CNST were observed in Chiapas and Guerrero (31.9 and 30.3); The incidence of neuroblastoma was low. Chiapas showed the highest incidence of retinoblastoma, bone tumors and carcinomas (21.8, 12.2 and 5.9 respectively); Puebla had the highest incidence of liver tumors (11.3) in which stages III and IV were the most frequent (67.8 %)., Conclusions: The main groups of cancer in Mexican children were leukemias, CNST, and lymphomas. Some states had high incidence; more than 50 % of children with solid tumors were diagnosed in advanced stages.

Smoking has a history almost as old as the own leaf of the tobacco, although in fact the act to smoke was restored in the general society after the discovery of America, considering itself even beneficial for the health, but fundamentally like a social and conventional conduct of the high nobility. In XVI century the first detractors began to appear but it was necessary many years to arrive the XX century when it was begun to relate the habit to tobacco and cancer, some years after the commercialization at industrial level of this habit. At the present the social awareness is tried towards the sanitary prevention. The declaration of Warsaw, the Marco Agreement and other sanitary measures forehead the habit to tobacco, are clear examples of this medical-social restlessness that tries to exile the smoking as a social, conventional and good seen conduct to try a prevention and a greater quality of life of the individuals.

Background: In 1996, Mexico started to register cases of childhood cancer. Here, we describe the incidence of cancer in children, residing in ten Mexican jurisdictions, who were treated by the Instituto Mexicano del Seguro Social (IMSS)., Methods: New cases of childhood cancer, which were registered prospectively in nine principal Medical Centers of IMSS during the periods 1998-2000 (five jurisdictions) and 1996-2002 (five jurisdictions), were analyzed. Personnel were specifically trained to register, capture, and encode information. For each of these jurisdictions, the frequency, average annual age-standardized incidence (AAS) and average annual incidence per period by sex and, age, were calculated (rates per 1,000,000 children/years)., Results: In total 2,615 new cases of cancer were registered, with the male/female ratio generally >1, but in some tumors there were more cases in females (retinoblastoma, germ cells tumors). The principal groups of neoplasms in seven jurisdictions were leukemias, central nervous system tumors (CNS tumors), and lymphomas, and the combined frequency for these three groups was 62.6 to 77.2%. Most frequently found (five jurisdictions) was the North American-European pattern (leukemias-CNS tumors-lymphomas). Eight jurisdictions had AAS within the range reported in the world literature. The highest incidence was found for children underless than five year of age. In eight jurisdictions, leukemia had high incidence (>50). The AAS of lymphomas was between 1.9 to 28.6. Chiapas and Guerrero had the highest AAS of CNS tumors (31.9 and 30.3, respectively). The frequency and incidence of neuroblastoma was low. Chiapas had the highest incidence of retinoblastoma (21.8). Germ-cell tumors had high incidence., Conclusion: The North American-European pattern of cancers was the principal one found; the overall incidence was within the range reported worldwide. In general but particularly in two jurisdictions (Yucatán and Chiapas), it will be necessary to carry out studies concerning the causes of cancer in children. Due to the little that is known about the incidence of cancer in Mexican children, it will be necessary to develop a national program to establish a cancer registry for the whole of the country.

The frequency of chromosomal alterations was compared among four children groups: those with Down syndrome and acute leukemia (DS/AL), those with acute leukemia (AL), those with only Down syndrome (DS) and healthy children (NC). The frequency of acquired chromosome abnormalities was larger in the AL group, followed by the DS/AL. The gaps and isogaps were more frequent in children with only DS. The polymorphisms of the constitutive heterochromatin were larger in the DS/AL group. These findings appear to imply that more genetic changes are necessary to develop AL in the case of healthy children compared to children with DS.

Unlabelled: Chagas disease is caused by the flagellate protozoan T: cruzi. Seroepidemiological surveys in Chiapas, Mexico have shown seropositive individuals, therefore, we searched for people affected by the chronic form of Chagas disease which involves the heart, causing a chronic, progressive and fatal disease called Chronic Chagasic Cardiopathy (CCC)., Material and Methods: To establish the frequency of CCC we studied 28 patients seen at the Hospital General Regional "Dr. Rafael Pascacio Gamboa" during October 2002 through October 2003 in Tuxtla Gutierrez, Chiapas, the State capital city, with diagnosis of dilated cardiomyopathy (DC), a serological survey for antibodies against T. cruzi was done. This hospital cares for people from all parts of Chiapas, Mexico. Clinical diagnosis of DC was established there and blind serological studies were performed in Mexico City., Results: Fifteen out of 28 DC patients (54%) had anti T. cruzi antibodies. All of them came from poor rural villages and they had heart failure and/or arrhythmia or heart blockade on EKG., Conclusions: This observation suggest that in Chiapas were Chagas disease is endemic, there are CCC patients. Any case with a clinical diagnosis of DC should be tested for antibodies against T. cruzi. The low socioeconomic status, culture and environment in this Mexican State favour the presence and transmission of this parasitic disease.

Sialolithiasis rarely occurs in children and usually affects the submandibular salivary gland. Because of its rarity, its clinical expression is frequently missed and diagnosis is delayed. We describe the case of a 9-year-old boy with submandibular sialolithiasis who presented spontaneous passage of a large salivary stone. We review the clinical findings and etiopathogenesis of sialolithiasis in children, as well as diagnostic imaging techniques and several therapeutic approaches to stone extraction.

Rickets usually occurs in the first two years of life and in puberty since metabolic demand is increased due to rapid growth in these two critical periods of life, when peak bone mass is achieved. Rickets remains one of the most prevalent pediatric diseases in developing countries. Although it is considered to have disappeared in developed countries, there is increasing evidence of widespread vitamin D deficiency among immigrants. There are many reports on rickets and osteomalacia in Asian infants, adolescents and pregnant women moving from India, Pakistan and Bangladesh to developed countries with a cooler climate. We describe three teenagers of Pakistani origin. Clinical presentation included limb pains, muscular weakness, knock-knees and seizures. In all three patients, biochemical findings included hypocalcemia, raised serum parathormone and alkaline phosphatase, and reduced 25-hydroxy vitamin D concentrations. After vitamin D treatment and dietary counseling, biochemical findings returned to normal and their symptoms improved. Given the recent increase in the number of immigrants to Spain, this forgotten disease will probably reappear.

Background: Exercise-induced asthma is defined as the transient broncho-spasm, that occurs after 3 to 8 minutes of continuous exercise; one of two mechanisms are implicated: the first is given by a hyper-osmolar environment interchange with the warm respiratory air and the water loss, the second due to reactive hyperemia or bronchial blood vessels edema., Objectives: To determine the effectiveness and safety of Zafirlukast treatment in exercise induce asthma, and in mild and moderated persistent bronchial asthma. Evaluate the security with laboratory test IL-2, IL-4, INFg, and CD69, to determinate TH1 and Th2 cells, laboratory and thorax x-ray determinations before and after zafirlukast treatment in exercise induce asthma, plus the functional respiratory test, and assert the clinical and adverse reaction with Zafirlukast., Material and Methods: A open, prospective, longitudinal study. Challenge test on a treadmill for 8 minutes. Twenty patients from the Allergy Service at Lic. Adolfo López Mateos Hospital, ISSSTE, in México City, fifteen female and five males. Aged 15 to 35 years. There was a control group of ten healthy subjects with similar age and sex. The drug Zafirlukast was administered 20 mg twice a day for eight weeks, with patient's informed and signed consent. Laboratory test: Blood Cell count, transaminases, bilirubins A, G, M and E immunoglobulins thorax X-ray, electrocardiogram, functional respiratory test before and after treatment., Results: Zafirlukast blocked exercise induced asthma in the early and late phases. There was a statistically significant improvement of a VEF-1 after exercise with a p > 0.001; furthermore, there was significant improvement in the mid-spiratory speed before the exercise with a p > 0.05. The mid-spiratory speed after the exercise, improved (p > 0.01). There were no collateral reactions, such as Churg-Strauss, only transitory headache in six and nauseas in two. There were no statistically significant changes in the cytokines assessment., Conclusions: There were no statistically significant changes in the cytokines assessments, in four cases the IL-4 decreased after the treatment. The anti-leukotriene improved the bronchospastic answer during the early and late phases, reducing the FEV-1, decreasing the recovering phase.

Background: American trypanosomiasis (Chagas' disease), an anthropozoonosis fairly common in rural Latin America, has become an urban disease due to continuous migration, intra- and internationally. Blood transfusion, the second important pathway for transmission, increases its impact. Recognition of seropositive subjects among blood donors is now recommended, and clinical and serological screening enforced. Maneuvers to inactivate or remove Trypanosoma cruzi present in collected blood are recommended., Methods: We surveyed voluntary donors at the National Institute of Cardiology in Mexico City in search of anti-T. cruzi by indirect immunofluorescence, ELISA, and Western blot analysis. Seropositive donors were identified and tested for immunoglobulin. We used types and fractions of donated blood to extract DNA and perform the PCR technique using kinetoplast primers seeking parasite DNA in blood., Results: After 3,300 donors were screened, we identified 10 seropositive subjects (0.3%). These subjects were considered as indeterminate chagasic patients, came mainly from rural areas, and had IgG (100%) and IgA (30%) antibodies against a crude extract as well as a recombinant T. cruzi antigen. Identification of parasite DNA in red cell and platelet fraction was achieved from eight blood units., Conclusions: The present data provide evidence that blood donors at an urban hospital are seropositive for T. cruzi and at least 50% of donors carry the parasite potentially able to transmit T. cruzi in their cellular blood products. Serological screening should be included in routine blood-making. It is also necessary to adopt measures to inactivate or eliminate organisms in donated blood.

Seven patients with diagnosis of dilated myocardiopathy, who fulfilled epidemiological, clinical, and serological criteria for chronic American Trypanosomosis, were submitted to hemoculture. Parasites were isolated in two patients (29%) confirming diagnosis and indicating an infection. Antibody titers did not show any relationship with the presence or absence of parasitemia nor were clinical differences noticed. Isolated Trypanosoma cruzi parasites belonged to biodeme III.

Chagas' disease (American Trypanosomiasis) affects more than 20 million people in Latin America. Almost 30% of those people may develop a chronic disease, which is expressed mainly as a chronic chagasic cardiopathy (CCC). Recent studies in Mexico have shown that 40% of patients suffering dilated cardiomyopathy do have serum antibodies against Trypanosoma cruzi. It is well known the difficulties of parasitologic diagnosis of CCC, which in less extent does exist for serologic diagnosis. Here we report a diagnostic method based on a molecular approach. It is able to recognize parasite DNA, and may have a clinical application. Two oligonucleotides (KNS1 and KNS2) designed from kinetoplast minicircle DNA, were used to amplify the hypervariable region by PCR technology. The method allowed an amplification of 0.8 to 1.5 minicircle DNA molecules, which equals 1/12,000 of parasite. When tissue DNA samples from mice infected with T. cruzi were subjected to amplification, a product was obtained that was recognized by a DNA probe specific for minicircle. These results correlate with immunohistochemical studies showing tissular parasites. Molecular diagnosis of American Trypanosomiasis, could be applied in human studies.