Development Of Related Diseases Research Articles

Metabolic syndrome: diagnostics and prevention in family doctor practice

The article presents the current data on the definition, diagnostic criteria, clinical and pathogenic characteristics of metabolic syndrome in adults and children in accordance with international guidelines and national standardized clinical protocols. The attention was paid to the correction the conditions, united by the term metabolic syndrome and preventive measures of development of related diseases in practice of a general practitioner – family physician. The features of drugs use (metformin) as a prophylactic measure were described.

Physical Training Status Determines Oxidative Stress and Redox Changes in Response to an Acute Aerobic Exercise.

Objective. To assess the influence of different physical training status on exercise-induced oxidative stress and changes in cellular redox state. Methods. Thirty male subjects participated in this study and were assigned as well-trained (WT), moderately trained (MT), and untrained (UT) groups. The levels of cortisol, creatine kinase, plasma reduced glutathione to oxidized glutathione (GSH/GSSG), cysteine/cystine (Cys/CySS), and GSH/GSSG ratio in red blood cells (RBCs) were measured immediately and 10 and 30 min after exercise. Results. Following the exercise, plasma GSH/GSSG (p = 0.001) and Cys/CySS (p = 0.005) were significantly reduced in all groups. Reduction in plasma GSH/GSSG ratio in all groups induced a transient shift in redox balance towards a more oxidizing environment without difference between groups (p = 0.860), while RBCs GSH/GSSG showed significant reduction (p = 0.003) and elevation (p = 0.007) in UT and MT groups, respectively. The highest level of RBCs GSH/GSSG ratio was recorded in MT group, and the lowest one was recorded in the WT group. Conclusion. Long term regular exercise training with moderate intensity shifts redox balance towards more reducing environment, versus intensive exercise training leads to more oxidizing environment and consequently development of related diseases.

Assessing correlations between geological hazards and health outcomes: Addressing complexity in medical geology

BackgroundThe field of medical geology addresses the relationships between exposure to specific geological characteristics and the development of a range of health problems: for example, long-term exposure to arsenic in drinking water can result in the development of skin conditions and cancers. While these relationships are well characterised for some examples, in others there is a lack of understanding of the specific geological component(s) triggering disease onset, necessitating further research. ObjectivesThis paper aims to highlight several important complexities in geological exposures and the development of related diseases that can create difficulties in the linkage of exposure and health outcome data. Several suggested approaches to deal with these complexities are also suggested. DiscussionLong-term exposure and lengthy latent periods are common characteristics of many diseases related to geological hazards. In combination with long- or short-distance migrations over an individual's life, daily or weekly movement patterns and small-scale spatial heterogeneity in geological characteristics, it becomes problematic to appropriately assign exposure measurements to individuals. The inclusion of supplementary methods, such as questionnaires, movement diaries or Global Positioning System (GPS) trackers can support medical geology studies by providing evidence for the most appropriate exposure measurement locations. ConclusionsThe complex and lengthy exposure–response pathways involved, small-distance spatial heterogeneity in environmental components and a range of other issues mean that interdisciplinary approaches to medical geology studies are necessary to provide robust evidence.

HLA-G 3142C/G polymorphism is related to development of symptoms in HTLV-1 infected individuals

The majority of HTLV-1 infected individuals remain asymptomatic (HAC) throughout life, and the risk factors associated to the development of related diseases, such as HAM/TSP and ATL, are not fully understood. The human leukocyte antigen-G (HLA-G) is expressed in several pathological conditions and it has been associated to immunosuppressive effects allowing the virusinfected cells to escape of the host antiviral defense. Here, we evaluated the correlation between HLA-G polymorphisms in symptomatic and asymptomatic HTLV-1 infected individuals. Four polymorphisms of the exon 8 of the HLA-G gene were analysed by gene sequencing in HAC (n=48) and HAM/TSP patients (n=43). No difference was found between the groups for 3035C/T, 3187A/G and 3196C/G polymorphisms. On the other hand, despite of the absence of statistical significance, HAC group showed higher frequency of the 3142GG genotype (responsible for decreased transcription of theHLA-G) compared to HAM/TSP group (p=0,0607). Still, HAC group showed lower frequency of the 3142CC genotype (responsible for increased transcription of the HLA-G) compared to HAM/TSP group (p=0,0762). Furthermore, HAC group showed statistically higher frequency of the 3142G allele and lower frequency of the 3142C allele compared to HAM/TSP group (p=0,0244). In conclusion, HTLV-1-related symptoms in HAM/TSP group could be partially determined by higher expression of HLA-G (caused by higher frequency of the 3142C allele). We hypothesise that the higher expression of HLA-G protects the HTLV-1 infected cell against immune system attack the increasing proviral load and trigger the HAM/TSP symptoms. Financial support FUNDHERP, FAPESP and CNPq.

Flavonoid Bioavailability and Attempts for Bioavailability Enhancement

Flavonoids are a group of phytochemicals that have shown numerous health effects and have therefore been studied extensively. Of the six common food flavonoid classes, flavonols are distributed ubiquitously among different plant foods whereas appreciable amounts of isoflavones are found in leguminous plant-based foods. Flavonoids have shown promising health promoting effects in human cell culture, experimental animal and human clinical studies. They have shown antioxidant, hypocholesterolemic, anti-inflammatory effects as well as ability to modulate cell signaling and gene expression related disease development. Low bioavailability of flavonoids has been a concern as it can limit or even hinder their health effects. Therefore, attempts to improve their bioavailability in order to improve the efficacy of flavonoids are being studied. Further investigations on bioavailability are warranted as it is a determining factor for flavonoid biological activity.

Investigation of microRNA expression changes in HepG2 cell line in presence of URG4/URGCP and in absence of URG4/URGCP suppressed by RNA interference

Hepatocellular carcinoma (HCC) originates from liver cells and is one of the most common malignant cancers in the world. microRNAs (miRNA), are single strand non-coding RNA molecules with the length of 18-25 nucleotides. miRNAs play an important role in the development of HCC, i.e., miRNAs have a significant impact on multistep hepatocellular carcinogenesis including cellular migration and invasion. URG4/URGCP (up-regulated gene-4/upregulator of cell proliferation) is up-regulated in the presence of HBxAg and has been identified and characterized by Satiroglu-Tufan et al. The full-length URG4/URGCP is 3.607 kb. Overexpression of URG4/URGCP in the presence of HBV X protein may function as a putative oncogene that significantly contributes to multi-step hepatocarcinogenesis. In this study, we aimed to investigate potential miRNA expression changes in HepG2 cell line model system in the presence of URG4/URGCP and in the absence of URG4/URGCP, which was suppressed by RNA interference. To functionally characterize URG4/URGCP, independent cultures of HepG2 cells were stably transfected with pcDNA3 or pcDNA3-URG4/URGCP. Relative quantification of whole genome miRNAs was analyzed by RT-PCR using human whole genome miRNA qPCR profiling kits. Among the 1,034 human miRNAs investigated by the arrays, 77 miRNAs were up-regulated and nine miRNAs were down-regulated in the presence of URG4/URGCP. In conclusion, we have analyzed miRNA profiles in HepG2 cells in presence or absence of URG4/URGCP gene using RNA interference. Some of these miRNAs may play roles in URG4/URGCP gene related disease development through the regulation of different signaling pathways.

Protective effect of anti‐TLR3 monoclonal antibody in Poly(I:C)/D‐galactosamine induced mouse sepsis model

Activation of toll‐like receptor 3 (TLR3) by foreign and endogenous ligands including viral dsRNA, bacterial RNA, mitochondrial RNA, and endogenous necrotic cell mRNA could trigger both apoptosis and inflammatory pathways that lead to disease development. Therefore, the hypothesis was that the blockade of TLR3 activity could be an approach to treat apoptosis and inflammation related disorders. First, anti‐TLR3 monoclonal antibodies were generated using standard hybridoma technology and then screened by Octet binding affinity assay. The antibody, R11H8, which specifically bound to mouse TLR3 receptor with Kd 2.97×10−10 M affinity was identified and expanded for cell‐based functional assays. Using TLR3‐overexpressing stable cell line, R11H8 dose‐dependently blocked poly(I:C)‐induced IL‐8 release (IC50 = 60nM). The mouse sepsis model was induced by administration of D‐galactosamine and poly(I:C). In this model, D‐galactosamine is a hepatotoxin which function as a sepsis sensitizer, and poly(I:C) is a sepsis‐inducing molecule that mimics dsRNA and activates TLR3. R11H8 (1 mg/mouse) was intraperitoneally injected into mice one day before Poly (I:C)/D‐galactosamine induction. Three days after sepsis induction, mice that did not receive the antibody were dead, whereas all of the mice in anti‐TLR3 treated group survived. The result confirmed that the blockade of TLR3 receptor activity could diminish the pathogenesis of dsRNA induced sepsis. The generation of anti‐TLR3 mAb, R11H8, provided a powerful tool for us to better understand the role of TLR3 in the inflammation and apoptosis related disease development.

Design, synthesis, and evaluation of an activity-based probe for cellular imaging of monoamine oxidases

Monoamine oxidase, a FAD-containing enzyme, is responsible for the degradation of various neurotransmitters and xenobiotic amines to aldehydes. The selective targeting of monoamine oxidases (MAOs) may also help to visualize the development of related diseases. A new activity-based fluorescent probe containing acetamide group has been synthesized. To minimize the interactions with the dye, a hexyl linker was inserted between MAO tracer 2-(alkylamino)-acetamide and fluorescein. Enzyme inhibitory activities of probe 1, compounds 2 and 3 were assayed to confirm a high binding affinity of the probe to MAOs (IC50 value: 35.1 μM for MAO-A and 150.8 μM for MAO-B). Cell imaging was investigated in breast cancer cell MCF-7 with confocal laser scanning microscopy, and the results showed that this new acetamide-based MAO probe has a high affinity and specificity toward MAO, and thus it is a promising candidate for sensitive MAO detection in living system.

A Proteomic Platform for EBV and KSHV Serological Screening

AbstractAbstract 1747Individuals with HIV infection have an increased susceptibility to EBV- and KSHV-associated malignancies. Evaluating a possible role for specific humoral responses to individual viral proteins in the control of viral load and prevention of EBV or KSHV related disease development in this population has been hampered by the limited number of viral antigens currently used for screening serum samples. We have developed a protein array platform that can be utilized for EBV and KSHV serological screening. The array consists of EBV and KSHV proteins printed onto glass slides. All eighty four EBV open reading frames and eighty six KSHV open reading frames were cloned into bacterial vectors and the inserts validated by DNA sequencing in both directions. Authenticated inserts were transferred to a yeast vector that expresses proteins as N-terminal GST-fusions. Seventy nine EBV proteins and eighty one KSHV proteins were successfully purified from yeast. First generation EBV/KSHV protein arrays have been printed with these proteins plus a variety of control proteins that include GST, and human IgG, IgM and IgA. Initial serological assays compared IgG antibody responses in HIV positive individuals, cancer patients and healthy adults. The HIV positive group (i) had a broader EBV antigenic response than the cancer patients or healthy normal's and (ii) recognized a wider range of EBV antigens compared to KSHV. This may reflect more extensive exposure to replicating EBV versus KSHV in these patients. In addition to the previously described dominant antigenic responses to EBV and KSHV, the assays identified multiple additional immunogenic viral proteins. The ability to compare serological responses to the complete repertoire of individual EBV and KSHV encoded proteins should provide new insight into B cell mediated immune regulation of these viruses. Disclosures:No relevant conflicts of interest to declare.

HLA-G 14-bp Insertion/Deletion Polymorphism Is a Risk Factor for HTLV-1 Infection

About 95% of HTLV-1 infected patients remain asymptomatic throughout life, and the risk factors associated with the development of related diseases, such as HAM/TSP and ATL, are not fully understood. The human leukocyte antigen-G molecule (HLA-G), a nonclassical HLA class I molecule encoded by MHC, is expressed in several pathological conditions, including viral infection, and is related to immunosuppressive effects that allow the virus-infected cells to escape the antiviral defense of the host. The 14-bp insertion/deletion polymorphism of exon 8 HLA-G gene influences the stability of the transcripts and could be related to HTLV-1-infected cell protection and to the increase of proviral load. The present study analyzed by conventional PCR the 14-bp insertion/deletion polymorphism of exon 8 HLA-G gene in 150 unrelated healthy subjects, 82 HTLV-1 infected patients with symptoms (33 ATL and 49 HAM), and 56 asymptomatic HTLV-1 infected patients (HAC). In addition, the proviral load was determined by quantitative real-time PCR in all infected groups and correlated with 14-bp insertion/deletion genotypes. The heterozygote genotype frequencies were significantly higher in HAM, in the symptomatic group, and in infected patients compared to control (p < 0.05). The proviral load was higher in the symptomatic group than the HAC group (p < 0.0005). The comparison of proviral load and genotypes showed that -14-bp/-14-bp genotype had a higher proviral load than +14-bp/-14-bp and +14-bp/+14-bp genotypes. Although HLA-G 14-bp polymorphism does not appear to be associated with HTLV-1 related disease development, it could be a genetic risk factor for susceptibility to infection.

Development of a proteomic platform for EBV and KSHV serological screening

In the context of HIV infection, there is an increased susceptibility to EBV- and KSHV-associated malignancies. Evaluating a possible role for specific humoral responses to individual viral proteins in the control of viral load and prevention of EBV or KSHV related disease development in this population has been hampered by the limited availability of suitable screening reagents. We have developed a protein array platform that can be utilized for EBV and KSHV serological screening. The array consists of EBV and KSHV proteins printed onto glass slides. All eighty-four EBV open reading frames and eighty-six KSHV open reading frames were cloned into bacterial vectors and the inserts validated by DNA sequencing in both directions. Authenticated inserts were transferred to a yeast vector that expresses proteins as N-terminal GST-fusions. Seventy-nine EBV proteins and eighty-one KSHV proteins were successfully purified from yeast. First-generation EBV/KSHV protein arrays have been printed with these proteins plus a variety of control proteins that include GST, and human IgG, IgM, and IgA. Glycosylation of viral proteins may affect immunogenicity. To evaluate this aspect, EBV and KSHV virion glycoproteins are being expressed in insect cells modified with the human glycosylation machinery. Once purified, these proteins will be added to the EBV/KSHV protein arrays. The ability to compare serological responses to the complete repertoire of individual EBV and KSHV encoded proteins should provide new insight into B cell mediated immune regulation of these viruses.

Reduced cardiotropic response to insulin in spontaneously hypertensive rats: role of peroxisome proliferator-activated receptor-γ-initiated signaling

Vascular insulin resistance plays a crucial pathogenic role in the development of genetic hypertension. However, it is not known whether hypertension-associated myocardial insulin resistance also exists, and whether this is involved in the development of related diseases, such as hypertensive heart failure. The present study aimed to determine whether hypertension-associated myocardial insulin resistance exists, in addition to any underlying mechanism. Ventricular myocytes were enzymatically isolated from male Wistar-Kyoto (WKY) rats or spontaneously hypertensive rats (SHR) and myocyte shortening and intracellular Ca2+ transient were assessed, as was any signaling mechanism. Compared with WKY rats, insulin-stimulated myocardial glucose uptake was blunted in SHR. More importantly, the positive inotropic effect of insulin was significantly reduced in SHR myocytes. Moreover, peroxisome proliferator-activated receptor (PPAR)gamma and phosphatidylinositol 3 (PI-3) kinase p85 expression and insulin-induced Akt phosphorylation were reduced in SHR cardiomyocytes, but were markedly restored when animals were treated with rosiglitazone for 14 days. Pretreatment with an Akt inhibitor abolished the inotropic effect induced by insulin in rosiglitazone-treated SHR cardiomyocytes. The data obtained demonstrate that insulin resistance exists in SHR cardiomyocytes as manifested by both reduced insulin-stimulated glucose uptake and an impaired contractile response to insulin, which is attributable to decreased PPARgamma expression and subsequent impairment in PI-3 kinase/Akt signaling.

T cell therapy of Epstein–Barr virus and adenovirus infections after hemopoietic stem cell transplant

Epstein–Barr virus (EBV)-associated post-transplant lymphoproliferative disease (PTLD) and adenovirus (AdV)-related pathologies are life-threatening complications of immunosuppression in recipients of hematopoietic stem cell transplantation (HSCT). In certain cohorts (unrelated and haploidentical donor HSCT, T-cell-depleted allograft), the risk of developing these complications is higher. Here we describe the impact of T cell therapy, within programs of specific routine surveillance and preemptive treatment, on the course of EBV infection, and development of related disease, in pediatric recipients of T-cell-depleted, HLA-haploidentical HSCT. Future prospectives include the transfer of this technology to treat AdV-related complications following HSCT.