Resistance exercise promotes functional test via sciatic nerve regeneration, and muscle atrophy improvement through GAP-43 regulation in animal model of traumatic nerve injuries.

Resistance training improves muscle strength through a combination of neural plasticity and muscle hypertrophy. This study aimed to evaluate the effects of resistance exercise on sciatic nerve regeneration and histology, growth-associated protein 43 (GAP-43) expressions, and soleus muscle atrophy following traumatic nerve injuries in Wistar rats. In the present study, 40 male Wistar rats were randomly assigned into four groups: healthy control (HC) as a sham group was exposed to the surgical procedures without any sciatic nerve compression, lesioned control (LC), resistance training (RT,non-lesioned), and lesioned rats + RT (LRT) (n = 10 in each). The RT group performed a resistance-training program 5 days/week for 4 weeks. Sciatic functional index (SFI) score, beam score and Basso, Beattie, and Bresnahan (BBB) score decreased and the hot plate time increased significantly in the LC| group compared to the HC (p < 0.05) group. However, the LRT| group showed a significant increase in the SFI score (p = 0.001) and a significant decrease in hot plate time (p = 0.0232) compared to the LC group. The LC group also showed neurological morphological damage and muscle atrophy and a decrease in GAP-43 in nerve tissue. In comparison to the LC group, a significant increase in sciatic nerve caliber, diameter, number of muscle fibers, and the expression of GAP-43| (p < 0.05) was observed in the LRT group. Doing resistance training even for four weeks seems to affect sciatic nerve lesions and injuries. It can also repair and regenerate nerve tissue by upregulating GAP-43 expression|, improving motor behavioral tests, and controlling muscle atrophy.

Diet Quality and Total Daily Price of Foods Consumed among Iranian Diabetic Patients.

BACKGROUND: The aim is to investigate the association between diet quality and daily price of foods consumed among Iranian diabetic patients. METHODS: This cross-sectional study was conducted among 200 patients with type 2 diabetes mellitus (T2DM) aged 30-70 years. General information, socioeconomic status, anthropometric and biochemical characteristics, and food prices were collected by pretested questionnaires. Dietary intakes were assessed using a semi-quantitative reliable and valid food frequency questionnaire. Modified nutritionist IV and SPSS software were used for analyses. RESULTS: The results of the present study indicated a direct relationship between total daily price of diet and nutrient adequacy ratio of Vitamin D, Vitamin B1, selenium, zinc, magnesium, potassium, and mean adequacy ratio of 11 micronutrients (Vitamin C, Vitamin E, Vitamin D, Vitamin B1, Vitamin B6, Vitamin B12, selenium, zinc, calcium, magnesium, and potassium) (P < 0.05). Furthermore, the total daily price of diet had a positive association with dietary intakes of protein, Vitamin D, Vitamin B1, selenium, zinc, magnesium and potassium among type 2 diabetic patients (P < 0.05). However, no significant relationship was observed between the total daily price of diet and anthropometric indices, biochemical characteristics, and socioeconomic status of participants in the present study (P > 0.05). CONCLUSIONS: This study showed that dietary quality and dietary intakes of energy, protein, and micronutrients were directly associated with the total daily price of foods among Iranian patients with type 2 diabetes.

Association of modified Nordic diet with cardiovascular risk factors among type 2 diabetes patients: a cross-sectional study.

Introduction: Cardiovascular disease (CVD) is one of the most important causes of mortality. Healthy diets can decrease CVDs and other chronic diseases especially in patients with type 2 diabetes. In this study, we investigate association between adherence to the modified Nordic diet and cardiovascular risk factors among patients with type 2 diabetes. Methods: This cross-sectional study was conducted among 339 type 2 diabetic patients. Anthropometric indices, blood pressure, and biochemical tests were evaluated. A validated and reliable semi-quantitative food frequency questionnaire (FFQ) was used to assess dietary intake. Nordic diet scores were calculated based on median intakes of six food groups. Results: Body mass index (BMI) was higher among participants who were in the lowest tertile of adherence to the Nordic diet (P=0.006). There was a significant association between socioeconomic status (SES) and adherence to the Nordic diet (P<0.0001). Participants who were in the top category of adherence to the diet had significantly lower levels of aspartate aminotransferase (AST) (P<0.0001). There was a significant inverse association between adherence to the Nordic diet and low density lipoprotein (LDL) levels (odds ratio [OR]=0.29 95% CI: 0.09, 0.91, P=0.025), high systolic blood pressure (SBP) levels (OR=0.35 95% CI=0.17-0.74, P=0.015), and risk of obesity (OR=0.25 95% CI: 0.10, 0.63, P=0.03). Conclusion: Results suggest that adherence to the Nordic diet is associated with reductions in the prevalence of obesity, LDL levels and blood pressure among type 2 diabetic patients. However, additional studies are needed to confirm these findings.

Antibody Response to Human Extracellular HER2 Subdomain Proteins in Mice.

BACKGROUND: In addition to passive immunotherapy using anti-HER2 monoclonal antibodies, active immunotherapy via HER2 targeting is an interesting approach to inducing specific anti-tumor immune responses. We have recently reported the immunogenicity of HER2 subdomains following DNA immunization and HER2 protein boosting. In the present study, we evaluated the immunogenicity of different HER2 extracellular subdomains for the induction of anti-HER2 antibody response in BALB/c mice. OBJECTIVE: To investigate and characterize antibody responses to human recombinant proteins of HER2 extracellular subdomains in immunized mice. METHODS: Four subdomains of HER2 extracellular domain were expressed in E.coli; subsequently, purified recombinant proteins were intraperitoneally injected in BALB/c mice with Freund's adjuvant. The anti-HER2 antibody response was detected by ELISA, immunoblotting and flow cytometry. RESULTS: All the four HER2 subdomains along with the full extracellular domain (fECD) were able to induce specific anti-HER2 antibodies. Although anti-HER2 subdomains antibodies could not react with eukaryotic recombinant fECD protein by ELISA, they were able to recognize this protein by immunoblotting under both reduced and non-reduced conditions. Furthermore, only the sera of mice immunized with fECD protein could recognize native HER2 on HER2 overexpressing tumor cells (>99%) by flow cytometry. Moreover, fECD immunized mice sera inhibited the proliferation of tumor cells by XTT assay. CONCLUSION: The prokaryotic recombinant proteins of HER2 extracellular subdomains are immunogenic, yet the induced specific antibodies do not react with the native HER2 protein due to the paucity of post-translation modifications and /or distortion of the native conformation of isolated HER2 extracellular subdomains which might be potentially effective for induction of cell mediated immune response against HER2.

Differential expression profiles of the salivary proteins SP15 and SP44 from Phlebotomus papatasi.

BACKGROUND: Sand fly saliva has been shown to help parasite establishment and to induce immune responses in vertebrate hosts. In the current study, we investigated the pattern of expression of two Phlebotomus papatasi salivary transcripts in specific physiological and seasonal conditions at a hyperendemic area of zoonotic cutaneous leishmaniasis (ZCL) in Iran. METHODS: Sand flies were collected during 2012-2013, and grouped according to physiological stages such as unfed, fed, semi-gravid, gravid, parous, nulliparous, infected or non-infected with Leishmania major and also based on the season in which they were collected. Quantitative Real-Time PCR was applied for assessment of the expression of two relevant salivary transcripts, PpSP15 and PpSP44, associated to protection from and exacerbation of ZCL, respectively. RESULTS: The expression of PpSP15 and PpSP44 transcripts was significantly up-regulated (1.74 and 1.4 folds, respectively) in blood fed compared to unfed flies. Among four groups of fed, unfed, semi-gravid and gravid flies, the lowest levels of PpSP15 and PpSP44 expression were observed in gravid flies. Additionally, the expression levels of both PpSP15 and PpSP44 transcripts in P. papatasi collected during summer were significantly up-regulated (3.7 and 4.4 folds, respectively) compared to spring collections. In addition, the PpSP15 transcript exhibited a significant up-regulation (P < 0.05) in non-infected flies compared to those infected with L. major. CONCLUSIONS: This study contributes to our knowledge of the differential expression of salivary genes among different groups within a P. papatasi population under natural field conditions. Cutaneous and visceral leishmaniasis are of public health importance in many parts of Iran and neighbouring countries where P. papatasi is the proven and dominant sand fly vector for ZCL, the most prevalent and endemic form of the disease in Iran. Therefore, the current study could be helpful in understanding the influence of salivary genes on Leishmania transmission by phlebotomine sand flies. Our findings demonstrate the differential expression of salivary transcripts under various physiological conditions potentially influencing the sand fly capacity for parasite transmission as well as the outcome of disease.

Seasonal and Physiological Variations of Phlebotomus papatasi Salivary Gland Antigens in Central Iran.

BACKGROUND: Sand fly saliva helps parasite establishment and induce immune responses in vertebrate hosts. In the current study, we investigated the modulation of Phlebotomus papatasi salivary gland antigen expression by seasonal and biological factors. METHODS: Sand flies were grouped according to physiological stages such as unfed, fed, semi-gravid, gravid, parous, nulliparous, infected or non-infected with Leishmania major and based on the season in which they were collected. Salivary gland antigens (SGAs) were analyzed using SDS-PAGE and the antibody response against SGAs in Rhombomys opimus was determined by ELISA and Western blot. RESULTS: The highest protein content was found in the salivary glands of unfed sand flies. The saliva content was higher in parous compared to nulliparous, in summer compared to spring, and in Leishmania-infected compared to non-infected flies. The salivary gland lysate (SGL) electrophoretic pattern variations were observed among sand flies with various physiological stages particularly from 4-9 protein bands of 14-70 kDa. The SGL of unfed and gravid flies had extra protein bands compared to fed and semi-gravid sand flies. There was missing protein bands in SGL of parous compared to nulliparous; and in summer compared to spring collected flies. Rhombomys opimus serum reacted strongly with an antigenic band of around 28 kDa in the SGL of all sand fly groups. CONCLUSION: Certain biological and environmental characteristics of wild populations of vector sand flies affect the protein content and antigenicity of saliva. This might have an important implication in the design of vector-based vaccines.

Antibody response to HER2 extracellular domain and subdomains in mouse following DNA immunization.

Human epidermal growth factor receptor 2 (HER2) is overexpressed in 15-20 % of breast cancer patients and is an appropriate target for immunotherapy in these patients. Monoclonal antibodies (mAbs) specific to HER2 are currently applied to treat breast cancer patients with HER2 overexpression. Active immunization with HER2 DNA or protein has been considered as a suitable alternative. The aim of this study is to evaluate anti-HER2 antibody response in serum of mice immunized with DNA constructs containing full extracellular domain (fECD) or subdomains of human HER2. Four extracellular subdomains and also fECD of HER2 were cloned into pCMV6-Neo vector. Different groups of Balb/C mice were immunized with HER2 DNA constructs and boosted with HER2 recombinant protein. The anti-HER2 antibody was subsequently determined by ELISA, flow cytometry, and immunohistochemistry. Anti-HER2 antibody was detected only in serum of mice immunized with fECD DNA. None of HER2 extracellular subdomains induced appreciable levels of anti-HER2 antibody. However, boosting with fECD or extracellular subdomain III (DIII) recombinant protein resulted in enhanced anti-HER2 fECD as well as anti-HER2 subdomain antibody responses. In this regard, almost all (99 %) of HER2-overexpressing BT474 cells could be detected by serum antibody from mice immunized with HER2 subdomain DNA and boosted with recombinant HER2 protein by flow cytometry. Similarly, serum of mice immunized with DIII DNA construct and boosted with recombinant DIII protein could also recognize these cells, but to a lesser extent (50 %). Our findings suggest that combination of HER2 DNA and protein immunization could effectively induce anti-HER2 antibody response in Balb/C mice.

Construction and Stable Expression of a Truncated Human Receptor Tyrosine Kinase Ror1 (Ror1-ECD).

Expression of receptor tyrosine kinase Ror1 in a wide variety of cancers has emerged as a new era focusing on targeting this receptor in cancer therapy. Our preliminary results indicate the presence of a truncated transcript of Ror1 in tumor cells. The truncated Ror1 encompasses extracellular and transmembrane domains, lacking catalytic kinase domain (Ror1-ECD). As enzyme activity is highly dependent on the catalytic domain, we were wondering how this transcript and its encoded protein could play a possible role in tumorigenesis. To understand the function of this truncated transcript and whether or not the encoded protein translocates to the cell surface, we constructed a mammalian expression vector containing exon 1 to exon 8 of human Ror1 gene as a model system. The encoded protein by this construct covers the entire extracellular and transmembrane domains of Ror1. The Chinese Hamster Ovary Cell line (CHO) was used for transfection. Our results showed that this construct could express Ror1-ECD at protein level and also the protein could effectively translocate to the surface of transfected cells. Such model may suggest that a proportion of Ror1 molecules expressed by tumor cells are not full-length Ror1. This notion may be considered when applying flow cytometry using antibodies against Ror1 for screening of tumor cells in order to avoid any miscalculation in the number of Ror1 molecules expressed by tumor cells. Furthermore, such expression may bring about assumptions on functional roles of Ror1-ECD in tumorigenesis, which requires extensive functional studies.

Docetaxel immunonanocarriers as targeted delivery systems for HER 2-positive tumor cells: preparation, characterization, and cytotoxicity studies.

BACKGROUND: The objective of this study was to develop pegylated poly lactide-co-glycolide acid (PLGA) immunonanocarriers for targeting delivery of docetaxel to human breast cancer cells. METHODS: The polyethylene glycol (PEG) groups on the surface of the PLGA nanoparticles were functionalized using maleimide groups. Trastuzumab, a monoclonal antibody against human epidermal growth factor receptor 2 (HER2) antigens of cancer cells, used as the targeting moiety, was attached to the maleimide groups on the surface of pegylated PLGA nanoparticles. Nanoparticles prepared by a nanoprecipitation method were characterized for their size, size distribution, surface charge, surface morphology, drug-loading, and in vitro drug release profile. RESULTS: The average size of the trastuzumab-decorated nanoparticles was 254 ± 16.4 nm and their zeta potential was -11.5 ± 1.4 mV. The average size of the nontargeted PLGA nanoparticles was 183 ± 22 nm and their zeta potential was -2.6 ± 0.34 mV. The cellular uptake of nanoparticles was studied using both HER2-positive (SKBR3 and BT-474) and HER2-negative (Calu-6) cell lines. CONCLUSION: The cytotoxicity of the immunonanocarriers against HER2-positive cell lines was significantly higher than that of nontargeted PLGA nanoparticles and free docetaxel.

Analysis of plasminogen activator inhibitor-1, integrin beta3, beta fibrinogen, and methylenetetrahydrofolate reductase polymorphisms in Iranian women with recurrent pregnancy loss.

PROBLEM: To identify the associations of the plasminogen activator inhibitor-1 (PAI-1) -675 4G/5G, beta fibrinogen (BF) -455G/A, integrin beta 3 (ITGB3) 1565T/C, and methylenetetrahydrofolate reductase (MTHFR) 677C/T and 1298A/C polymorphisms with recurrent pregnancy loss (RPL). METHOD OF STUDY: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were performed to assess the frequency of five candidate genetic risk factors for RPL, and the frequencies of the polymorphisms were calculated and compared between case and control groups. RESULTS: The BF -455G/A, MTHFR 677C/T, and 1298A/C polymorphisms were found to be positively, and ITGB3 1565T/C polymorphism negatively, associated with RPL. Homozygosity but not heterozygosity for PAI-1 -675 4G/5G polymorphism was significantly higher in patients with RPL than in the control group. The presence of both mutations of MTHFR genes highly increased the risk of RPL. CONCLUSION: The data highlight the importance of thrombophilia screening in patients with RPL.

Peptide-based Polyclonal Antibody Production against P110 Protein of Mycoplasma genitalium.

Mycoplasma genitalium (M.genitalium) is a sexually transmitted pathogen. Detection of this microorganism in clinical specimens by culture is rather difficult and time consuming. The aim of this study was to produce polyclonal antibody against a synthetic peptide from P110 protein of M.genitalium in order to develop a diagnostic tool for detection of this microorganism in clinical specimens. A synthetic peptide from P110 protein was conjugated to Keyhole Limpet Hemocyanin (KLH) and used for immunization of a white New Zealand rabbit. The produced antibody was purified by affinity chromatography and its specific interaction with immunizing peptide was determined by ELISA. Immunoreactivity of the antibody was also tested by Western blotting in bacterial cell lysate prepared from M.genitalium G-37. To confirm its application as a diagnostic tool, indirect immunofluorescent staining method was performed on M.genitalium-infected PBMC using anti-P110 as the primary antibody. The results showed that produced antibody has excellent reactivity with immunizing peptide and also detected a single band of 110 kDa corresponding to P110 protein. M.genitalium-infected PBMC showed a bright fluorescent signal in IF staining. This antibody might be used as a tool in diagnostic applications.