Socioeconomic Level and the Relationship in Glycemic Behavior in the Mexican Population. A Nutritional Alternative Focused on Vulnerable Populations.

In this study, numerical approximations were generated to analyze the behavior of glycemic variations in the non-diabetic population of the Mexican republic. The main objective of this work is to obtain an overview of the glycemic variations in the non-diabetic population from different socioeconomic statuses in Mexico (Medium-high, medium, and low). Thus, evaluating the effect on the glucose level under a healthy diet considering the socioeconomic capabilities of the population. Through the national health and nutrition survey of Mexico 2020 and the Mexican food base, 1420 virtual patients were proposed (522 low status, 485 medium status and 413 Medium-High status) focused on simulating the glycemic behavior in each of the survey participants. Considering that the average food expenditure of the Mexican population is $107.00 MXN, and the cost of a healthy diet is $66.50 MXN, the economic sustainability of the Mexican population to adopt a healthy diet is revealed. The particularity of this work is focused on obtaining diverse data that are difficult to access in the development of population analyses. Such is the case of the approach proposed for different socioeconomic statuses. In this way, the proposed methodology provides a framework for complementary research contributions to the subject.

Deformed wing virus type a and b in managed honeybee colonies of Argentina.

Apis mellifera is infected by more than 24 virus species worldwide, mainly positive-sense, single-stranded RNA viruses of the Dicistroviridae and Iflaviridae families. Among the viruses that infect honey bees, Deformed wing virus is the most prevalent and is present as three master variants DWV-A, B, and C. Given that the ectoparasitic mite Varroa destructor vectors these virus variants, recombination events between them are expected, and variants and their recombinants can co-exist in mites and honeybees at the same time. In this study, we detect, through RT-qPCR, the presence of DWV-A and B in the same samples of adult bees from colonies of Argentina. Total RNA was extracted from pools of ten adult bees from 45 apiaries distributed across the main beekeeping Provinces of Argentina (Buenos Aires, Santa Fe, Córdoba, Santiago del Estero, Río Negro, and Mendoza); then RT-qPCR reactions were performed to detect DWV-A and B, with specific primer pairs. After the amplifications, PCR products (204 and 660 bp amplicons for DWV-B, and ~250 bp for DWV-A) were purified and sequenced to verify that they corresponded to reported sequences, analyzing them using the Blast software. Of the 45 samples analyzed by RT-qPCR, over 90% were infected with DWV-A and 47% were also positive for DWV-B, where it was found in high prevalence specifically in colonies of A. mellifera of the Buenos Aires Province. Future studies will determine the impact of this type of the virus and its ability to recombine with the other DWV types in the apiaries of our country.

Transgenic Sweet Orange expressing hairpin CP-mRNA in the interstock confers tolerance to citrus psorosis virus in the non-transgenic scion.

The lack of naturally occurring resistance to citrus psorosis virus (CPsV) necessitates a transgenic approach for the development of CPsV-resistant citrus. To evaluate the feasibility of conferring resistance to a non-transgenic scion, we have assembled citrus plants by grafting combining a non-transgenic Sweet Orange as scion, CPsV-resistant transgenic Sweet Orange lines expressing intron-hairpin (ihp) RNA derived from the viral coat protein (ihpCP) as interstock, and a non-transgenic citrus as rootstock. We demonstrated that ihpCP-transcripts translocate through the graft from interstock to scion, triggering the silencing of coat protein mRNA target. Two independent CPsV challenge assays showed that expression of ihpCP in the interstock provides resistance against CPsV in the interstock, and different levels of protection in the non-tg scion, depending of the virus delivery site. These results indicated that grafting is a promising biotechnological alternative to protect woody plants against virus infections in vegetative propagated plants.

RAS mutation analysis in circulating tumor DNA from patients with metastatic colorectal cancer: the AGEO RASANC prospective multicenter study.

Background: RAS mutations are currently sought for in tumor samples, which takes a median of almost 3 weeks in western European countries. This creates problems in clinical situations that require urgent treatment and for inclusion in therapeutic trials that need RAS status for randomization. Analysis of circulating tumor DNA might help to shorten the time required to determine RAS mutational status before anti-epidermal growth factor receptor antibody therapy for metastatic colorectal cancer. Here we compared plasma with tissue RAS analysis in a large prospective multicenter cohort. Patients and methods: Plasma samples were collected prospectively from chemotherapy-naive patients and analyzed centrally by next-generation sequencing (NGS) with the colon lung cancer V2 Ampliseq panel and by methylation digital PCR (WIF1 and NPY genes). Tumoral RAS status was determined locally, in parallel, according to routine practice. For a minimal κ coefficient of 0.7, reflecting acceptable concordance (precision ± 0.07), with an estimated 5% of non-exploitable data, 425 subjects were necessary. Results: From July 2015 to December 2016, 425 patients were enrolled. For the 412 patients with available paired plasma and tumor samples, the κ coefficient was 0.71 [95% confidence interval (CI), 0.64-0.77] and accuracy was 85.2% (95% CI, 81.4% to 88.5%). In the 329 patients with detectable ctDNA (at least one mutation or one methylated biomarker), the κ coefficient was 0.89 (95% CI, 0.84-0.94) and accuracy was 94.8% (95% CI, 91.9% to 97.0%). The absence of liver metastases was the main clinical factor associated with inconclusive circulating tumor DNA results [odds ratio = 0.11 (95% CI, 0.06-0.21)]. In patients with liver metastases, accuracy was 93.5% with NGS alone and 97% with NGS plus the methylated biomarkers. Conclusion: This prospective trial demonstrates excellent concordance between RAS status in plasma and tumor tissue from patients with colorectal cancer and liver metastases, thus validating plasma testing for routine RAS mutation analysis in these patients. Clinical Trial registration: Clinicaltrials.gov, NCT02502656.

Cervical cancer cells suppress effector functions of cytotoxic T cells through the adenosinergic pathway.

BACKGROUND: The expression of CD73 in tumor cells plays a significant role in the production of adenosine (Ado) that suppresses antitumor effector cells. METHODS: In this study we analyzed the capability of HPV-positive (HPV+) cervical cancer (CeCa) cell lines CaSki, SiHa, HeLa, and RoVa; and HPV-negative (HPV-) cell lines C33A and ViBo to produce Ado and inhibit effector functions of CD8+ T cells. RESULTS: HPV+ CeCa cells expressed significantly higher levels of CD73 in the membrane (p<0.01) than HPV- CeCa cells and this expression was associated with the production of larger amounts of Ado (>400µM) compared to HPV-CeCa cells (<200µM) in the presence of AMP, as well asa stronger inhibition of (>50%) proliferation, activation, and cytotoxic activity of CD8+ T cells via interaction with A2A adenosine receptor. We also provide evidence that silenced E6/E7 expression in CeCa cells, strongly reduced its CD73 expression level and its capability to generate Ado. CONCLUSION: This results suggest that HPV infection, which is associated with more than 99% of CeCa cases, may present an increased constitutive expression of CD73 in cervical neoplasia to contribute to the suppression of the immune response mediated by the production of large amounts of Ado.

Citrus psorosis virus coat protein-derived hairpin construct confers stable transgenic resistance in citrus against psorosis A and B syndromes.

Citrus psorosis virus (CPsV) is the causal agent of psorosis, a serious and widespread citrus disease. Two syndromes of psorosis, PsA and PsB, have been described. PsB is the most aggressive and rampant form. Previously, we obtained Pineapple sweet orange plants transformed with a hairpin construct derived from the CPsV coat protein gene (ihpCP). Some of these plants were resistant to CPsV 90-1-1, a PsA isolate homologous to the transgene. In this study, we found that expression of the ihpCP transgene and siRNA production in lines ihpCP-10 and -15 were stable with time and propagation. In particular, line ihpCP-15 has been resistant for more than 2 years, even after re-inoculation. The ihpCP plants were also resistant against a heterologous CPsV isolate that causes severe PsB syndrome. Line ihpCP-15 manifested complete resistance while line ihpCP-10 was tolerant to the virus, although with variable behaviour, showing delay and attenuation in PsB symptoms. These lines are promising for a biotech product aimed at eradicating psorosis.

Exposure to high ambient temperatures alters embryology in rabbits.

High ambient temperatures are a determining factor in the deterioration of embryo quality and survival in mammals. The aim of this study was to evaluate the effect of heat stress on embryo development, embryonic size and size of the embryonic coats in rabbits. A total of 310 embryos from 33 females in thermal comfort zone and 264 embryos of 28 females in heat stress conditions were used in the experiment. The traits studied were ovulation rate, percentage of total embryos, percentage of normal embryos, embryo area, zona pellucida thickness and mucin coat thickness. Traits were measured at 24 and 48 h post-coitum (hpc); mucin coat thickness was only measured at 48 hpc. The embryos were classified as zygotes or two-cell embryos at 24 hpc, and 16-cells or early morulae at 48 hpc. The ovulation rate was one oocyte lower in heat stress conditions than in thermal comfort. Percentage of normal embryos was lower in heat stress conditions at 24 hpc (17.2%) and 48 hpc (13.2%). No differences in percentage of zygotes or two-cell embryos were found at 24 hpc. The embryo development and area was affected by heat stress at 48 hpc (10% higher percentage of 16-cells and 883 µm2 smaller, respectively). Zona pellucida was thicker under thermal stress at 24 hpc (1.2 µm) and 48 hpc (1.5 µm). No differences in mucin coat thickness were found. In conclusion, heat stress appears to alter embryology in rabbits.

Correlated response in litter size components in rabbits selected for litter size variability.

A divergent selection experiment for the environmental variability of litter size (Ve) over seven generations was carried out in rabbits at the University Miguel Hernández of Elche. The Ve was estimated as the phenotypic variance within the female, after correcting for year-season and parity-lactation status. The aim of this study was to analyse the correlated responses to selection in litter size components. The ovulation rate (OR) and number of implanted embryos (IE) in females were measured by laparoscopy at 12 day of the second gestation. At the end of the second gestation, the total number of kits born was measured (TB). Embryonic (ES), foetal (FS) and prenatal (PS) survival were computed as IE/OR, TB/IE and TB/OR, respectively. A total of 405 laparoscopies were performed. Data were analysed using Bayesian methodology. The correlated response to selection for litter size environmental variability in terms of the litter size components was estimated as either genetic trends, estimated by computing the average estimated breeding values for each generation and each line, or the phenotypic differences between lines. The OR was similar in both lines. However, after seven generations of selection, the homogenous line showed more IE (1.09 embryos for genetic means and 1.23 embryos for phenotypic means) and higher ES than the heterogeneous one (0.07 for genetic means and 0.08 for phenotypic means). The probability of the phenotypic differences between lines being higher than zero (p) was 1.00 and .99, respectively. A higher uterine overcrowding of embryos in the homogeneous line did not penalize FS; as a result, this line continued to show a greater TB (1.01 kits for genetic means and 1.30 kits for phenotypic means, p = .99, in the seventh generation). In conclusion, a decrease in litter size variability showed a favourable effect on ES and led to a higher litter size at birth.

Application of horizontal-flow anaerobic immobilized biomass reactor for bioremediation of acid mine drainage.

The production of low-pH effluent with sulfate and metals is one of the biggest environmental concerns in the mining industry. The biological process for sulfate reduction has the potential to become a low-cost solution that enables the recovery of interesting compounds. The present study analyzed such a process in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor, employing ethanol as the carbon and energy source. Results showed that a maximal efficiency in the removal of sulfate and ethanol could only be obtained by reducing the applied sulfate load (225.1 ± 38 g m(-3) d(-1)). This strategy led to over 75% of chemical oxygen demand (COD) and sulfate removal. Among the COD/SO4(2-) studied ratios, 0.67 showed the most promising performance. The effluent's pH has naturally remained between 6.8 and 7.0 and the complete oxidation of the organic matter has been observed. Corrections of the influent pH or effluent recirculation did not show any significant effect on the COD and sulfate removal efficiency. Species closely related to strains of Clostridium sp. and species of Acidaminobacter hydrogenomorfans and Fusibacter paucivorans that can be related to the process of sulfate reduction were found in the HAIB reactors when the initial pH was 5 and the COD/SO4(2-) ratio increased to 1.0.

Recurrent wheezing and asthma after bocavirus bronchiolitis.

BACKGROUND: Human bocavirus (HBoV) was recently discovered and identified as an important cause of respiratory infection in young children. However, the relationship between HBoV-bronchiolitis and the development of recurrent wheezing has not yet been established. OBJECTIVE: We designed this study in order to describe the mid-term outcome, regarding the development of recurrent wheezing and asthma of HBoV-bronchiolitis patients and to compare it with RSV-bronchiolitis infants. METHODS: We studied 80 children (10 with HBoV and 70 with RSV infection), currently aged ≥4 years and previously hospitalised during the seasons 2004-2009 due to acute bronchiolitis. Epidemiological and clinical data were collected through structured clinical interviews at the follow-up visit. Spirometry and skin prick tests to common food and inhaled allergens were performed. RESULTS: All HBoV-patients developed recurrent wheezing and half of them had asthma at age 5-7 years. Almost 30% required hospital admission for recurrent wheezing. Asthma (odds ratio (OR)=1.28) and current asthma (OR=2.18) were significantly more frequent in children with HBoV-bronchiolitis than in RSV-bronchiolitis. FEV1 values were 99.2±4.8 in HBoV-group vs. 103±11 in RSV-group, p: 0.09. No differences were found with respect to allergic rhinitis, atopic dermatitis, food allergy, proportion of positive prick tests, and family history of atopy or asthma. CONCLUSIONS: Severe HBoV-bronchiolitis in infancy was strongly associated with asthma at 5-7 years.

The Salmonella enterica serovar Typhi ltrR-ompR-ompC-ompF genes are involved in resistance to the bile salt sodium deoxycholate and in bacterial transformation.

A characterization of the LtrR regulator, an S. Typhi protein belonging to the LysR family is presented. Proteomics, outer membrane protein profiles and transcriptional analyses demonstrated that LtrR is required for the synthesis of OmpR, OmpC and OmpF. DNA-protein interaction analysis showed that LtrR binds to the regulatory region of ompR and then OmpR interacts with the ompC and ompF promoters inducing porin synthesis. LtrR-dependent and independent ompR promoters were identified, and both promoters are involved in the synthesis of OmpR for OmpC and OmpF production. To define the functional role of the ltrR-ompR-ompC-ompF genetic network, mutants in each gene were obtained. We found that ltrR, ompR, ompC and ompF were involved in the control of bacterial transformation, while the two regulators and ompC are necessary for the optimal growth of S. Typhi in the presence of one of the major bile salts found in the gut, sodium deoxycholate. The data presented establish the pivotal role of LtrR in the regulatory network of porin synthesis and reveal new genetic strategies of survival and cellular adaptation to the environment used by Salmonella.

Mesenchymal stromal cells derived from cervical cancer tumors induce TGF-ß1 expression and IL-10 expression and secretion in the cervical cancer cells, resulting in protection from cytotoxic T cell activity.

Cervical cancer (CeCa) tumors are characterized by increased expression of TGF-ß1 and IL-10, which are correlated with downregulated expression of major histocompatibility complex class I antigens (HLA-I) on cancer cells and a reduced immune response mediated by cytotoxic T lymphocytes (CTLs). Mesenchymal stromal cells (MSCs) are important components in the tumor microenvironment that have been suggested to contribute to cancer progression through the induction of TGF-ß1 and IL-10. In this study, we provided evidence that MSCs derived from cervical tumors (CeCa-MSCs) cocultured with CeCa cells induced significant expression of TGF-ß1 and secretion of IL-10 by CeCa cells compared to MSCs derived from the normal cervix (NCx-MSCs) and normal bone marrow (BM-MSCs; gold standard). This increase in expression was associated with a significant downregulation of HLA-I molecules and protection of the cells against specific CTL lysis. Interestingly, the addition of the neutralizing antibody anti-TGF-ß to the CeCa/CeCa-MSCs coculture strongly inhibited the expression and production of IL-10 by CeCa cells. Anti-TGF-ß as well as anti-IL-10 also abolished HLA-I downregulation, and reversed the inhibition of CTL cytotoxicity. These results provide evidence that TGF-ß1 and IL-10 could play an important role in the downregulation of HLA-I molecules on CeCa cells induced by tumor MSCs. Our findings suggest a novel mechanism through which MSCs may protect tumor cells from immune recognition by specific CTLs.

Multidisciplinary Approach in Hepatocellular Carcinoma in a Level II Hospital: The First Decade of Hospital Universitario Fundacion Alcorcon.

BACKGROUND/AIMS: Mean survival in hepatocellular carcinoma remains low. Many efforts have been done during the last years through screening, diagnosis and treatment to improve the results. The aim of this work is to present the experience of our hospital multidisciplinary group during the first decade of this century. METHODOLOGY: The patients with hepatocellullar carcinoma presented at the multidisciplinary meeting from 1999 to 2009 were prospectively studied. According to the tumor and functional status they were treated through the current available guidelines by transplant, partial hepatectomy, local/regional procedures, systemic or symptomatic treatment. RESULTS: One hundred and forty two patients were studied. Median tumor size was 3 cm. A single tumor was diagnosed in 64.8% of the patients. Eighteen patients had liver resection (6 transplantation and 12 with partial resection), 53 tumors were not treated due to advanced stage or liver dysfunction, and in the remaining patients radiofrequency, ethanol or embolization treatments were used, single or combined. CONCLUSIONS: a multidisciplinary approach of hepatocellular carcinoma in a second level hospital with trained professionals permits a diagnosis in early tumoral and functional stages in the majority of patients, and a variety of possible treatments with adequate survival outcomes.

Hospital admission due to respiratory viral infections in moderate preterm, late preterm and term infants during their first year of life.

BACKGROUND: Respiratory viral infections are a major cause of hospitalisation in infants <1 year and might cause severe symptoms in preterm infants. Our aim was to analyse admissions due to respiratory infections in moderate, late and term infants, and to identify risk factors for hospitalisation in preterm versus term. METHODS: Prospective study in a cohort of moderate and late preterm, and term infants born between October/2011 and December/2012. Admissions due to respiratory infections during the first year of life were analysed and compared among moderate (32-33), late (34-36) and term infants. Sixteen respiratory viruses were detected by RT-PCR. Clinical data were collected. RESULTS: 30 (20.9%) out of 143 preterm infants required admission for respiratory infection, versus 129 (6.9%) of 1858 term infants born in the same period (p<0.0001, OR: 3.6 CI 2.0 to 5.0). Hospitalised children had a higher prevalence of hyaline membrane disease (HMD) at birth (p<0.001, OR: 7.7 CI: 2.121 to 27.954) and needed more mechanical ventilation (p<0.001, OR: 5.7 CI: 1.813 to 18.396). Virus was identified in 25/30 (83%) preterm babies, and in 110/129 (85%) term infants. The most frequent viruses in preterm infants were RSV (76%) rhinovirus (20%). Clinical and epidemiological characteristics among term and preterm infants were similar. CONCLUSIONS: The risk of respiratory admissions during the first year of life is up to 3.6 times higher in moderate and late preterm. Once admitted, clinical features of respiratory episodes requiring hospitalisation are similar among term and preterm infants. Hyaline membrane disease and mechanical ventilation were also risk factors for respiratory admissions.

Antioxidant, phase II and III responses induced by lipoic acid in the fish Jenynsia multidentata (Anablapidae) and its influence on endolsulfan accumulation and toxicity.

Antioxidants like lipoic acid (LA) are known to trigger augmented antioxidant and phase II and III responses. This study aimed to evaluate the effect of LA in P-glycoprotein (Pgp) expression, glutathione-S-transferase (GST) activity, total antioxidant competence, levels of lipid peroxides (TBARS) and accumulation of the organochlorine insecticide endosulfan (Endo: α-, ß-isomers and sulfate metabolite) in different organs of the fish Jenynsia multidentata. One hundred and twenty females (1.55±0.07 g) were fed during 8 days with (n=60) or without (n=60) a LA enriched ration (6000 mg/kg). Four experimental groups were defined: -LA/-Endo; +LA/-Endo; -LA/+Endo; and +LA/+Endo. Endo groups were exposed during 24 h to 1.4 µg of insecticide/L. Results showed that only LA induced a significant increment in liver Pgp expression. GST activity was augmented in liver after exposure to LA or Endo. TBARS levels were lowered in liver and gills after LA pre-treatment. Total antioxidant capacity was lowered in liver of Endo exposed fish, a result that was reversed by LA pre-treatment. It is concluded that LA induced the expected effects in terms of Pgp expression, GST activity and reduced TBARS levels although favored α-Endo accumulation in brain. However, the Endo metabolism to the more persistent endosulfan sulfate was not facilitated by LA pre-treatment.

Novel nanostructured lipid carriers loading Apigenin for anterior segment ocular pathologies.

Dry eye disease (DED) is a chronic multifactorial disorder of the ocular surface caused by tear film dysfunction and constitutes one of the most common ocular conditions worldwide. However, its treatment remains unsatisfactory. While artificial tears are commonly used to moisturize the ocular surface, they do not address the underlying causes of DED. Apigenin (APG) is a natural product with anti-inflammatory properties, but its low solubility and bioavailability limit its efficacy. Therefore, a novel formulation of APG loaded into biodegradable and biocompatible nanoparticles (APG-NLC) was developed to overcome the restricted APG stability, improve its therapeutic efficacy, and prolong its retention time on the ocular surface by extending its release. APG-NLC optimization, characterization, biopharmaceutical properties and therapeutic efficacy were evaluated. The optimized APG-NLC exhibited an average particle size below 200 nm, a positive surface charge, and an encapsulation efficiency over 99 %. APG-NLC exhibited sustained release of APG, and stability studies demonstrated that the formulation retained its integrity for over 25 months. In vitro and in vivo ocular tolerance studies indicated that APG-NLC did not cause any irritation, rendering them suitable for ocular topical administration. Furthermore, APG-NLC showed non-toxicity in an epithelial corneal cell line and exhibited fast cell internalization. Therapeutic benefits were demonstrated using an in vivo model of DED, where APG-NLC effectively reversed DED by reducing ocular surface cellular damage and increasing tear volume. Anti-inflammatory assays in vivo also showcased its potential to treat and prevent ocular inflammation, particularly relevant in DED patients. Hence, APG-NLC represent a promising system for the treatment and prevention of DED and its associated inflammation.

Combined HER2 analysis of biopsies and surgical specimens to optimize detection of trastuzumab-eligible patients in eso-gastric adenocarcinoma: a GERCOR study.

BACKGROUND: HER2 is overexpressed in 10 to 20% of gastro-esophageal adenocarcinoma (GE-ADK), and is a target for trastuzumab in metastatic patients. We conducted a study to compare HER2 expression between diagnostic biopsies (DBs) and surgical specimens (SSs) of GE-ADK, and to determine the influence of non-trastuzumab containing neoadjuvant chemotherapy (NAC) on this expression. PATIENTS AND METHODS: Pathological specimens from biopsies of 228 patients operated on between 2004 and 2011 were collected. Two cohorts treated (n = 141) or not (n = 87) with a NAC were constituted. Two blind independent pathological HER2 analyses on DB and on SS were carried out using immunohistochemistry (IHC) and colorimetric in situ hybridization (CISH). HER-2 overexpression (HER2+) was defined by a score 3+ in IHC, or 2+ with a positive CISH test, according to the specific HER2 scoring guidelines for GE-ADK. RESULTS: Paired HER2 status could be determined for 218 out of the 228 patients (95.6%). HER2+ rates were 13.3% on DB (29/218) and 14.7% on SS (32/218). HER2+ tumors were mainly cardial or esophageal adenocarcinomas, with a well-differentiated, intestinal histological type. HER2 status differed between DB and SS in 6% of cases. When DB analyses were added to SS analyses, the relative increase in HER2+ cases was 13.5% (17.1% for patients with NAC and 23.5% for patients with histological response to NAC, versus 7.1% for patients without NAC, P = 0.4, NS). Differences between DB and SS HER2 expression could be explained by intratumoral heterogeneity and by a HER2 expression decrease in SS after NAC in responding patients possibly due to a higher chemosensitivity of HER2-positive clones. CONCLUSION: The determination of HER2 status on DB provides results that complete those obtained with SS. Combining the analysis of DB and of SS enables to optimize the selection of trastuzumab-eligible patients in case of metastatic relapse, and particularly in previously NAC-responding patients.

First Report of Tomato rugose yellow leaf curl virus Infecting Tomato in Argentina.

Tomato plants exhibiting typical symptoms of begomovirus infection, including leaf deformation, curling, and yellowing, were collected from cultivated fields in Lavalle Department, Corrientes, Argentina, in 2010. Although the number of affected plants was only 2% within a farm, the finding is of considerable importance since the white fly Bemisia tabaci is widely spread within the country, even in other southernmost areas such as the cinturón hortícola de Buenos Aires (horticultural belt around Buenos Aires). DNA isolated from infected tomato leaves collected from three symptomatic tomato plants was amplified by PCR with specific primers designed to amplify a region of component A and B of the Begomovirus genome (3). The amplified DNA fragment was sequenced and a new set of primers were designed based on the obtained sequences. A DNA fragment of about 1,300 bp was amplified and later the complete genome, which was 2,683 bp long. No fragments were obtained when template DNA was from non-infected leaf samples. The 2,683-bp fragment was annotated at the NCBI under Accession No. KC132844. Analysis by NCBI BLAST showed that it was highly homologous to DNA-A component of Begomovirus. Furthermore, the genome organization was typical of DNA-A component of bipartite New World begomovirus. The sequence had one open reading frame (ORF) on the viral-sense strand (AV1/CP) and four ORFs on the complementary-sense strand (AC1/Rep, AC2/TrAp, AC3/REn, and AC4). In order to confirm this finding, the viral genome was amplified by rolling circle amplification (RCA, TempliPhi 100 Amplification Kit, Amersham Biosciences) as described by the manufacturer instructions. The RCA full-length product was digested with XhoI generating a 2,700-bp DNA fragment, suggesting the presence of only one restriction site, in agreement with the bioinformatics analysis of the KC132844 sequence. This PCR product was used as template in PCR reactions with specific primers to DNA-A or DNA-B components. While the DNA-A primers generated the expected 1,300-bp fragment, those homologous to the DNA-B component did not generate amplifications. These results confirmed the identity of the DNA-A component of the isolate MT8. The full sequence of the DNA-A component was 94% homologous to the DNA-A sequence of the Uruguayan begomovirus Tomato Rugose Yellow Leaf Curl Virus-[U4.1] (JN381823.1). Therefore, considering our results and the criteria proposed by Fauquet (1), isolate MT8 is a new species of begomovirus described recently (2). This is the first report of TRYLCV in one of the main areas of tomato production in Argentina. This virus might be accompanying another begomovirus TYVSV that provoked yellow veins symptoms in tomato plants cultivated in the same area of Corrientes. These viruses appeared recently and concomitantly with the introduction of the white fly Bemisia spp. in the area, which is one of the main production areas of tomato and provides fresh tomatoes to the whole country, and in wintertime to the city of Buenos Aires, when the horticultural belt around Buenos Aires is not under production. References: (1) C. M. Fauquet et al. Arch Virol 153:783, 2008. (2) B. Márquez-Martín et al. Arch Virol 157:1137, 2012. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.

A novel lipid nanocarrier for insulin delivery: production, characterization and toxicity testing.

A novel nanocarrier based on solid lipid nanoparticles (SLNs) was developed for insulin delivery using a novel double emulsion method. Physical stability of particles was assessed by size analysis using dynamic light scattering (DLS), matrix crystallinity by differential scanning calorimetry (DSC) and toxicity analysis by Drosophila melanogaster testing. Insulin-SLNs were composed of Softisan®100 1.25% wt, Lutrol®F68 1% wt, soybean lecithin 0.125% wt, and loaded with 0.73-0.58 mg/mL peptide. Placebo-SLNs (insulin-free) also contained 0.025% wt Tween®80. Mean particle sizes of placebo-SLN and insulin-SLN were 958 ± 9.5 and 978 ± 8.3 nm, respectively. The polydispersity index (PI) was 0.28 ± 0.018 and 0.29 ± 0.013, respectively. Polarized light microscopy analysis depicted no aggregation of developed particles. DSC analysis allowed characterizing SLN with 43-51% matrix crystallinity. Using Drosophila melanogaster test, no toxicity was reported for SLN and for the bulk lipid. This study shows that SLNs are promising and helpful to overcome conventional insulin therapy, in particular for their lack of toxicity for oral delivery.

[Unexpected diagnosis of nephronopthisis in the genetic study of hypertension due to histological diagnosis of benign nephroangioesclerosis evolved in a young caucasian patient]. / Diagnóstico inesperado de nefronoptisis en estudio genético de hipertensión por diagnóstico histológico de nefroangioesclerosis benigna evolucionada en paciente joven caucásico.

We present the case of a young Caucasian patient with renal disease of unclear cause, with a final diagnosis of advanced benign nephroangiosclerosis established by renal biopsy. Due to the possibility of having hypertension in pediatric age (without study or treatment), with the renal biopsy findings, the genetic study showed polymorphisms risk in the APOL1 and MYH9, and also an unexpected diagnosis of a complete deletion of the NPHP1 gene in homozygosis, associated with the development of nephronophthisis. In conclusion, this case illustrates the importance of carrying out a genetic study in youngs patients with renal disease unclear cause, even having a histological diagnosis of nephroangiosclerosis.