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Background Peripheral neuropathy is one of the most common late complications of diabetes. Vascular endothelial growth factor (VEGF) gene polymorphisms have been associated with the development of peripheral neuropathy in different populations of patients with type 2 diabetes mellitus (DM2). Objective To analyze the prevalence of the +936 C/T VEGF gene polymorphism among patients with DM2 with and without peripheral neuropathy. Study design and methodology 218 unrelated DM2 patients, 90 with and 128 without peripheral neuropathy were genotyped for the +936 C/T VEGF gene polymorphism using PCR amplification followed by restriction length polymorphism analysis. Results The CC homozygous VEGF+936 C/T (rs3025039) was the predominant genotype in DM2 patients with peripheral neuropathy, whereas the predominant genotype in patients without neuropathy was the heterozygous C/T. No statistical association was found between genotype distribution and the presence of neuropathy (p = 0.063). The distribution of the genotypes according to the dominant (CC vs. CT + TT) and recessive (TT vs. CT + CC) models showed that the homozygous CC and TT genotypes, respectively, are not risk factors for neuropathy. The CT genotype conferred a protective effect as seen in the over-dominant model (CT vs. CC + TT) (OR = 0.52; 95% CI = 0.300–0.90; p = 0.019). Conclusion We conclude that the VEGF+936 C/T (rs3025039) gene polymorphisms are related to peripheral neuropathy in Mexican DM2 patients, with the heterozygous genotype potentially conferring a protective effect.

Background The rambutan (Nephelium lappaceum L) is an exotic fruit native to the Southeast Asia and currently cultivated in different tropical areas, among them in southern Mexico. It is characterized by its nutritionals and functionals properties and has been relevant for its commercialization and acceptance in different countries and Mexico represents one of the main producers and exporter of rambutan in the world. Scope and approach In this review, we summarize information about of the bioactive compounds present in the rambutan fruit, together with the nutritionals properties that it presents as a functional food, with a focus on its by-products (pulp, seed and peel). The rambutan fruit contains important bioactive compounds, mainly the peel has a high content of antioxidants which are of interest in the food, cosmetic and pharmaceutical industries. Key findings and conclusions Extracts of rambutan fruit, mainly from the peel, have been shown to possess phytochemical compounds that exhibit antioxidant, antimicrobial, antidiabetic, antiviral, anti-inflammatory, anti-hypoglycemic and anti-proliferative effects in various in vitro and in vivo tests. However, it is necessary to further analyze the nutritional and functional potential of this fruit, the therapeutic mechanisms involved and to develop its industrial process as a functional or nutraceutical food product.

Obesity is a complex, chronic, and multifactorial disease that has become a major, and worldwide, public health problem contributing to an increased number of pathologies, including type 2 diabetes, cardiovascular disease, hyperlipidemia, and metabolic syndrome, thus suggesting a commolon origin. A diet high in sugar and fats coupled with a sedentary lifestyle has a major role in the development of obesity. However, the genetic background has also been associated with body fat accumulation. The aim of this study was to assess the effect ofACE-rs4646994, APOA5-rs662799, and MTP-rs1800591 gene polymorphisms on clinical and biochemical parameters and to evaluate the association with body phenotypes in children and adolescent population of Saltillo, Coahuila, Mexico.Anthropometric, clinical, biochemical parameters and BMI were obtained from 405 children and adolescents. The BMI was used to determine the body phenotype. The rs4646994 gene polymorphism was determined by PCR, whereas rs662799 and rs1800591 were determined by PCR-RFLP. The obtained results were analyzed to determine their association of these single nucleotide polymorphisms with body phenotype and biochemical parameters.TT genotype for APOA5-rs662799 was associated with increased levels of HDL-C in the analyzed population (p0.05). The ACErs4646994gene polymorphism is associated with high Insulin levels, HOMAIR index, and triglyceride levels, mainly when presenting a I/I genotype (p0.05).The polymorphic allele of the ACE gene is capable of modulating triglyceride levels, insulin levels and HOMA-IR index in the evaluated population; it must be highlighted that this has not been reported in other studied populations elsewhere.

Different strategies were assessed for the production of ethanol from Agave lechuguilla that was pretreated by autohydrolysis. Separate hydrolysis and fermentation (SHF) was compared against simultaneous processes including simultaneous saccharification and fermentation (SSF) and prehydrolysis and simultaneous saccharification and fermentation (PSSF) using different solids (15%, 20%, and 25% w/w) and enzyme loadings (15 FPU/g, 20 FPU/g, and 25 FPU/g glucan). The results showed that the maximum ethanol concentration (53.7 g/L) and productivity (1.49 g/L h-1) was obtained at 36 h in the SHF configuration at the highest solids and enzyme loadings (25% w/v and 25 FPU/g glucan, respectively). The ethanol concentration and productivity obtained in the PSSF configuration at the same time were 45 g/L and 1.25 g/L h-1, respectively. The SSF configuration exhibited the lowest ethanol concentration and productivity (10.4 g/L and 0.29 g/L h-1, respectively) at 36 h. The enzyme used, Cellic CTec3, allowed for high glucose yields at the lower enzyme dosage assessed. The SHF configuration exhibited the best results. However, the PSSF configuration can be considered an attractive alternative because it eliminated the need for solid-liquid separation devices, which simplifies the industrial implementation of the process.

Pectin methylesterase is an enzyme with an important in vivo role in plants as well as in food industry. This enzyme catalyzes the hydrolysis of methyl ester bounds in pectin which is one of the main components of cell wall in plants, producing methanol and free carboxylic groups. The effect of pectin methylesterase in food quality has been extensively studied, producing desirable effects in texture improvement as well as undesirable effects in some beverages. Likewise, the low methoxyl pectin produced by this enzyme has characteristics that contribute to formulate best quality food products. Pectin methylesterase is a ubiquitously enzyme that presents multiple isoforms, but is not only present in plants; it is also found in fungi, bacteria, and yeast, which have specific chemical and physical characteristics. The latter makes the task of analyzing the wide variety of these enzymes with its specific characteristics difficult. Based on this enzyme relevance and the aforementioned, multiple methods have been developed in order to evaluate pectin methylesterase activity with different research objectives. In this paper, the importance of the enzyme as well as advantages and drawbacks of the different methods will be discussed besides applications and evolution of these will be mentioned. Additionally, this paper will improve the understanding of the systems used in pectin methylesterase activity analysis.

The aim of this study was to evaluate the fermentation of dietary fiber from green bean (Phaseolus vulgaris) and prickly pear shell (Opuntia ficus-indica) by Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum 450B growing as mono-culture and co-culture, the fermentation products, and proteins expressed during this process. The analysis of the fermentation profile showed a major growth of bacteria in the culture media of each dietary fiber supplemented with glucose, and particularly B. bifidum 450B at 48 h showed the highest growth. In the case of the co-culture, the growth was lower indicating the possible negative interaction between L. acidophilus LA-5 and B. bifidum 450B and may be due to the less amount of carbohydrates and the high content of non-soluble fiber that affected the nutrients availability for the bacterial strains. The pH changes indicated the presence of short-chain fatty acids (SCFAs), being acetate (46-100%) the main SCFA. Changes in the proteome concerned proteins that are involved in carbohydrate and other carbohydrate pathways. The characterization of the bacteria according to the growth, metabolites, and proteins expressed allows understanding the response to the change of environmental conditions and could be useful to understand L. acidophilus LA-5 and B. bifidum 450B strains' adaptation to specific applications.

Anaerobic digestion (AD) is a cost-effective treatment for management of lignocellulosic substrates, viz., agricultural wastes and animal manures, which also aids in generation of methane as biofuel. Although the application of AD technology is increasing, one of the major limitations of the process is that the rate of fermentation is higher than the rate of methanogenesis, which significantly affects process stability and methane yield. Normally, the souring of digesters can be observed after 2–4 weeks after the initiation of the volatile fatty acids accumulation, which makes it difficult for early detection and consequently resulting in acidification of digesters. Of late, metagenomic approaches are gaining importance due to their ability to reveal the microbial diversity and their dynamics in a relatively short time. However, their functional nature could not be clearly explained due to the lack of data on their activity. Recent advances in proteomic studies show its potential as a complementary technology to metagenomic studies for efficient management of digesters. Metaproteomic analyses aid in identifying a shift in metabolic paths and in metabolic networks under stress conditions. This provides insights on functionality, microbial interactions, and provides data on spatiotemporal variations and their dynamics of proteins crucial for efficient performance of the digester. Besides, this technique has led to identify novel phylotypes with novel functions among the microbial communities of the anaerobic digesters, which suggest the potential of proteomics in bioprospection of novel enzymes for industrial purposes. How proteomics along with metagenomics and transcriptomics data could aid in early detection of disturbances in the digesters helps in formulating recovery strategies as well as to increase the methane content of biogas will be discussed in this chapter.

Moringa oleifera is a rich source of polyphenols, but its content and profile may vary according to environmental conditions, harvest season, and plant tissue. This study was carried out in order to determine the phenolic compounds by HPLC/MS, as well as, the in vitro antioxidant activity of extracts from leaflets and petioles of Moringa oleifera planted in three locations. The content of total polyphenolic compounds was estimated. In addition, hydrolyzable polyphenols were determined using the Folin-Ciocalteu technique, while, condensed polyphenols were estimated employing the butanol-HCl method. These chemical compounds were identified by high-performance liquid chromatography coupled to masses spectrometry (HPLC/MS). In vitro antioxidant activity of the extracts was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylenebenzothiozoline-6-sulfonic acid (ABTS) and the ferric reducing antioxidant power (FRAP) methods. The polyphenolic compounds were found mainly in the leaflets. The HPLC/MS allowed the identification of 15 compounds of the hydroxycinnamic acid, anthocyanin, flavonols, methoxyflavone, tyrosol, dihydrochalcones, flavones, and methoxyflavone families. The ethanolic extracts showed high antioxidant activity for DPPH, ABTS, and FRAP, especially, those obtained from the leaflets produced at Ramos Arizpe, Coahuila. The amounts of condensed were higher than those of hydrolyzable polyphenols in the moringa grew at three environments. While the highest amount of condensed polyphenols was observed in moringa grew at Ramos Arizpe. Thus, the presence of these polyphenol compounds may explain the great antioxidant activity (98.45% inhibition) showed with the ABTS radical, in the moringa ethanolic extracts from Ramos Arizpe. These results indicate that moringa may have great potential as a natural antioxidant agent.

The rambutan peel (RP) is a relevant source of bioactive molecules, which could be used for application in cosmetics, food, and pharmaceutical areas. Total soluble polyphenol content was extracted from Mexican variety rambutan peels using an emergent ultrasound/microwave-assisted extraction (U/M-AE) technology. Five extractions were performed using different mass/volume and ethanol/water ratios, 1:16-0, 1:16-70, 1:8-0, 1:8-70, 1:12-30. Condition 1:16-0 was defined as the best extraction condition with 0% ethanol percentage (only water). The content of total soluble polyphenols was 307.57 mg/g. The total bound polyphenol content was 26.53 mg/g. Besides, two separation processes were made with the soluble fraction, the first one was performed using Amberlite XAD-16 (Sigma-Aldrich, Saint Louis, MO, USA), and seven polyphenolic compounds were obtained. The second one was performed using a preparative HPLC (Varian, Palo Alto, CA, USA) equipment obtained fraction where three compounds were obtained: geraniin (main compound), ellagic acid, and ellagic acid pentoside. The major compound isolated in the two separations was geraniin, according to HPLC/ESI/MS (High Performance Liquid Chromatography/ElectroSpray Ionization/Mass) analysis.

Fructose and fructo-oligosaccharides (FOS) are important ingredients in the food industry. Fructose is considered an alternative sweetener to sucrose because it has higher sweetening capacity and increases iron absorption in children, and FOS's are a source of dietary fiber with a bifidogenic effect. Both compounds can be obtained by enzymatic hydrolysis of inulin. However, inulin presents limited solubility at room temperature, thus, fructose and FOS production is carried out at 60°C. Therefore, there is a growing interest to isolate and characterize thermostable inulinases. The aim of this work was to evaluate the capacity of different fungal strains to produce potential thermostable inulinases. A total of 27 fungal strains belonging to the genera Aspergillus, Penicillium, Rhizopus, Rhizomucor and Thermomyces were evaluated for production of inulinase under submerged culture using Czapek Dox medium with inulin as a sole carbon source. Strains were incubated at 37°C and 200 rpm for 96 h. Crude enzyme extract was obtained to evaluate inulinase and invertase activity. In order to select the fungal strain with the highest thermostable inulinase production, a selection criterion was established. It was possible to determine the highest inulinase activity for Rhizopus microsporus 13aIV (10.71 U/mL) at 36 h with an optimum temperature of inulinase of 70°C. After 6 h at 60°C, the enzyme did not show any significant loss of activity and retained about 87% activity, while it only retains 57% activity at 70°C. According to hydrolysis products, R. microsporus produced endo and exo-inulinase.

Hepatitis C virus (HCV) infection causes high level of oxidative stress and hepatic inflammation primarily, and derives in different pathologies as liver cirrhosis, hepatic failure, and hepatocellular carcinoma (HCC). Currently available treatment is a combination of pegylated interferon-α and ribavirin, surgical resection, and liver transplantation. However, more than 50% of patients remain diseased and the treatment prior mentioned causes serious adverse effects and there is still no preventive vaccine available. Considering the limited treatment options, it remains the urgent need to find additional therapeutic agents. Chemoprevention is defined as the use of nontoxic substances and it has been considered as the alternative strategy to reduce the cancer development and carcinogenesis before invasion and metastasis occur. Polyphenols, such as ellagic acid, ellagitannins, and punicalagin and phenolic acids, such as gallic acid are abundant plant phenolic compounds and they are extracted from different fruit peels and seed. Those kinds of compounds are known to reduce the risk of disease and have shown to induce cytotoxicity in several cancer cell lines, including liver cell lines. In vitro and in vivo studies have shown that the GA has a positive effect on various liver diseases, such as hepatitis by HCV infection, hepatocellular carcinoma, steatosis, and fatty liver. Histopathology and serum biochemical studies in mice fed with a diet supplemented with GA, observed that GA protects against hepatic steatosis, obesity, hypercholesterolemia, and insulin resistance. This effect occurs in part through a reversing of the common high fat diet that caused disturbances to a range of metabolic pathways, including lipid metabolism, glucose metabolism (glycolysis and gluconeogenesis), and amino acids metabolism. On the other hand, GA also has in vitro effect in hepatocarcinoma cells that express nonstructural proteins of HCV reducing viral replication. Viral proteins expression and the levels of oxidative stress decreases the viability of liver cancer cells in cell culture. Some of the main action mechanisms are the activation of protein kinase ATM, dehydrogenase UDP-glucose, and NFκB pathway. In addition, when glutathione is depleted, induction of apoptotic, antiangiogenic effect, and inhibition of tumor cell proliferation is activated. For these reasons, the use of antioxidant compounds, particularly gallic acid, has been studied recently because it is a naturally abundant plant phenolic compound, it may be considered a tool as a biomarker, or alternative therapy in the prevention and treatment of liver diseases.

Pseudomonas putida strain (HM346961) was isolated from a consortium of bacteria acclimatized to unleaded gasoline-contaminated water. The consortium can efficiently remove benzene, toluene, ethylbenzene and xylene (BTEX) isomers, and a similar capability was observed with the P. putida strain. Proteome of this strain showed certain similarities with that of other strains exposed to the hydrocarbon compounds. Furthermore, the toluene di-oxygenase (tod) gene was up-regulated in P. putida strain when exposed to toluene, ethylbenzene, xylene, and BTEX. In contrast, the tod gene of P. putida F1 (ATCC 700007) was up-regulated only in the presence of toluene and BTEX. Several differences in the nucleotide and protein sequences of these two tod genes were observed. This suggests that tod up-regulation in P. putida strain may partially explain their great capacity to remove aromatic compounds, relative to P. putida F1. Therefore, new tod and P. putida strain are promising for various environmental applications.

Inulinase is an enzyme produced by plants and several microorganisms, including fungi, to hydrolyze the β-2,1 glycosidic linkages present in some oligosaccharides to produce fructose and glucose. This enzyme, in conjunction with invertases, levanases, and two types of 1-fructosyl transferases have been described as members of the glycosyl hydrolases (family 32), the most diverse group of enzymes used by microbes for biomass degradation. As being part of the same clan, they have common evolutionary origin sharing the most important functional characteristics. Recently, a xerophylic fungi strain isolated from Mexican semi-desert, Penicillium citrinum ESS has been reported as inulinase producer, which could have greater stability than other enzymes due to a metabolic machinery adapted to typical temperature changes in this region. To continue the understanding of action mechanisms of these enzymes and to establish evolutionary relationships within this family, in the present study, phylogenetic analyses were used to analyze amino acid sequences coding fungal and yeast glycoside hydrolases of family 32, including the new sequenced inulinase of P. citrinum ESS. It was possible to elucidate the action mechanism of fungal glycoside hydrolases in present study and to classify inulinase from P. citrinum ESS as an exo-inulinase on the basis of their amino acid sequence phylogenetic affinities.

Background and aim. Acetylsalicylic acid (ASA) has been shown to downregulate HCV expression; however, the involved mechanisms are unknown. We used proteomic analysis to compare protein expression profiles between human hepatocarcinoma cells (Huh7) and Huh7-HCV cells harboring expression of non-structural HCV proteins, to elucidate the mechanism(s) involved in ASA-mediated downregulation of HCV replication. Material and methods. Both cell lines were treated or untreated with 4 mM ASA and harvested at 0, 24, 48 and 72 h to isolate total proteins, which were resolved by two-dimensional gel electrophoresis (2DE) to separate them by isoelectric point (pI), followed by fractionation by molecular weight (MW). Gels were scanned and analyzed with PD-Quest software V8.0.1, and proteins were elucidated by the specific pI and MW using TAGIDENT software. Statistics analysis included the t-test. Results and Discussion. Different protein patterns among hepatocytes expressing HCV-proteins in ASA treated and untreated cells were found. Among proteins differentially expressed in Huh7-HCV cells, we found proteins related to cell proliferation (MTMR6, FAM22, HDGF and HCF-1) after 24 h of ASA treatment; and upregulation of angiostatin, PI4KA and STAT-1 after 48 h of treatment. Finally, at 72 h of ASA exposure, we identified overexpression of adenylsuccinate synthase, 2’-3’-di-deoxyadenosine, ubiquitin-protein-ligase E6A, adenylosuccinate-lyase and nibrin (NBN). Conclusion. We found that ASA induces different protein patterns in Huh7-HCV cells promoting activation of proteins involved in cell progression, repair of double strand breaks, proliferation, inhibition of apoptosis and growth stimulation at the same time that it decreased HCV expression.

In this study, we evaluated the efficiency of BTEX biodegradation by a consortium acclimatized to unleaded gasoline and bacterial strains isolated from it. The isolates were identified by PCR-amplification of 16S rRNA genes, and BTEX biodegradation was confirmed by the identification of dioxygenase-related proteins. The consortium degraded 95% of total BTEX and the following genera were present in the consortium: Pseudomonas, Shewanella, Burkholderia, Alcanivorax, Rhodococcus, and Bacillus. Strain FMB08, isolated from a consortium, was capable of removing 90% of total BTEX and its 16S rDNA analysis was similar to that of Pseudomonas.

Development of multidrug-resistant bacteria (MDRB) and demand by consumers of fresher foods with no chemical preservatives added, and minimally processed with long shelf life, have increased demand for new antibacterial compounds, which has stimulated the identification, development, and redesign of antibiotics. For this reason, different antibacterial agents, including antimicrobial peptides (AMPs) and bacteriocins are now being reevaluated. The latter compounds are peptides produced by different bacteria, which may act against a wide range of pathogens. Some of them are closely related to spoilage and disease-causing Gram-positive bacteria. In this chapter, we reviewed the use of bacteriocins for the control of MDRB. Bacteriocins have different applications in the food industry, especially for biopreservation and shelf-life extension, also as clinical antimicrobials and as microbial control during fermentation. There are different types of bacteriocins successfully applied in the food industry, such as nisin and pedicin PA-1/AcH, which have been authorized to be used as bio-preservatives. But they are only a small fraction of the diversity of these bacteriocins. Bacteriocins are a good option to use against multidrug-resistant (MDR) pathogens in clinical cases because these AMPs may act at picomolar and nanomolar concentrations and with high specificity. Continuous research will lead to obtaining more diverse and efficient antimicrobial bacteriocins; in this case, recent development of new techniques for identification, purification, and heterologous proteins will allow for new antibiotics that can control MDRB.

Zearalenone (ZEA) is a non-steroidal estrogenic mycotoxin produced by a variety of Fusarium fungi that are commonly found in feed. ZEA cause reproductive disorders of farm animals and occasionally hyperoestrogenic syndromes of humans; also, has been shown to be hepatotoxic, genotoxic, hematotoxic and immunotoxic, evidence primarily based on studies with different biological models in vitro and in vivo in poultry and laboratory rodents. However, there is little knowledge about of the effects of ZEA in human leukocytes. Objective: The aim of this study was to investigate the cell viability and lipid peroxidation effects caused by ZEA in human peripheral blood leukocytes. Methods: Human leukocytes were exposed in vitro with ZEA in concentrations ranging from 10 to 80 μg/mL during 1 h of incubation. Cell viability was measured by staining with trypan blue and neutral red assays, and lipid peroxidation was evaluated by the amount of the lipid peroxidation product, malondialdehyde (MDA). Results: ZEA significantly decreased the cell viability in human leukocytes with concentrations ranging from 10 μg/mL to 80 μg/mL. Also, we found significant increases of MDA levels in the cells exposed to concentrations between 40 and 80 μg/mL. Conclusion: The data suggest that lipid peroxidation is involved in the citotoxicity of ZEA in human leukocytes.

Diabetic retinopathy (DR) is one of the primary causes of blindness in the working age population and is characterized by angiogenesis in the retina. Platelets have been suggested to be involved in the pathogenesis of diabetic microvascular complications. The integrin receptor for collagen/laminin, α2β1, mediates platelet primary adhesion to subendothelial tissues, which is an essential first step in thrombus formation. The gene encoding the α2 subunit of α2β1 integrin has ≥8 polymorphisms, including a BglII/NdeI restriction fragment length polymorphism. To explore the prevalence of DR in a population from Northeastern Mexico, unrelated, hospitalized patients who had received a diagnosis of type 2 diabetes mellitus (DM2) at least 10 years previously were recruited (n=177). DR was diagnosed in a masked manner by independent ophthalmologists using fundus images captured using a non-mydriatic retinal camera. A total of 121 patients with DM2 (68%) had some degree of DR development (DR patients), and 56 patients with DM2 (32%) did not exhibit any sign of DR (No-DR patients). The results showed that after 15 years of DM2 progression, there is an increased risk of DR (P=0.0497; odds ratio, 1.993). In addition, insulin therapy and family history of DM2 were significantly associated with DR. In order to detect a possible association between DR and BglII/NdeI α2 gene polymorphisms, a comparative cross-sectional study between DR and No-DR patients was conducted. The α2 gene was genotyped by polymerase chain reaction-restriction fragment length polymorphism assay. Statistical analysis revealed no association between BglII/NdeI genotypes and the development of DR in this group of patients. In conclusion, the present data indicate a high prevalence of DR in the Mexican population and suggest that the damage in DR is due to other factors, such as the duration of the DM2, and is not linked to BglII/NdeI α2 gene polymorphisms.