RESUMEN

BACKGROUND: Primary dysmenorrhea is the usual medical status in medical students that are defined as pain
during the menstrual period. This study was done to evaluate the psychological problems associated with dysmenorrhea.
Materials and Methods: Three hundred forty students aged 18 to 20 years participated in this cross­sectional
study (194 with dysmenorrhea and 150 without dysmenorrhea). In this cross­sectional study, data were collected
through the sociodemographic checklist, the verbal multidimensional scoring system (VMS), and the revised
version of the Symptom Checklist-90 (SCL-90-R) questionnaire using the convenience sampling method. This
questionnaire includes 9 Subscale and a GSI index. We considered psychological distress to be equivalent to
the Global Severity Index (GSI), which is obtained by dividing 90 questions by 90. The significance level of the
tests was considered 0.05.
Results: The GSI of the SCL-90 score in the 194 students with dysmenorrhea and 150 students without dysmenorrhea
was 1.02 ± 0.42 and 0.34 ± 0.15 respectively (P<0.001). In the group with dysmenorrhea, the severity
of dysmenorrhea was significantly associated with a family history of dysmenorrhea and mother's education
(P=0.012 and P=0.037, respectively). The strongest predictors of GSI>1 were a family history of dysmenorrhea
and mother's education [odds ratio (OR)=2.33, 95% confidence interval (CI), 1.43-4.15 and OR=0.45, 95% CI,
0.24-0.87, respectively].
Conclusion: According to the result, dysmenorrhea is associated with psychological distress. Psychological interventions
and counseling in addition to drug treatment are suggested for treatment of primary dysmenorrhea.
Therefore, it is necessary to formulate strategies and health policies to recover psychological issues of menstrual
health.

RESUMEN

CONTEXT: Breast cancer is the most common cancer in women worldwide and its prevalence is increasing. AIMS: The aim of this study was to investigate the extent to which a history of infertility can present as risk factors for breast cancer. SETTINGS AND DESIGN: This was a hospital-based case-control study. SUBJECTS AND METHODS: In this study, 1177 women with breast cancer were participated for assessing the risk for this cancer. The control was 1204 women with self-reported free-cancer history who were matched in terms of age and residence. STATISTICAL ANALYSIS USED: Using logistic regression, it was examined whether infertility is a risk factor of case-control status in addition to marriage age, menarche age, body mass index, number of pregnancies, family history of breast cancer, and previous oral contraceptive use. The data were considered significant at P ≤ 0.05. RESULTS: Infertility history was reported in 12.5% (n = 147) of the cases and 5.8% (n = 70) of the controls. Infertility history was a relevant risk for case-control study in addition to other risk factors (odds ratio: 2.43; 95% confidence interval, 3.34-1.77). CONCLUSIONS: This study found that infertility may be as the main risk factor for breast cancer in Iranian women, therefore, doing breast screening in women who have one or more risk factors, must receive more emphasis.

Asunto(s)

RESUMEN

A hybrid biological-biomaterial antigen delivery vector comprised of a polymeric shell encapsulating an Escherichia coli core was previously developed for in situ antigen production and subsequent delivery. Due to the engineering capacity of the bacterial core, the hybrid vector provides unique opportunities for immunogenicity optimization through varying cellular localization (cytoplasm, periplasm, cellular surface) and type (protein or DNA) of antigen. In this work, three protein-based hybrid vector formats were compared in which the pneumococcal surface protein A (PspA) was localized to the cytoplasm, surface, and periplasmic space of the bacterial core for vaccination against pneumococcal disease. Furthermore, we tested the hybrid vector's capacity as a DNA vaccine against Streptococcus pneumoniae by introducing a plasmid into the bacterial core to facilitate PspA expression in antigen presenting cells (APCs). Through testing these various formulations, we determined that cytoplasmic accumulation of PspA elicited the strongest immune response (antibody production and protection against bacterial challenge) and enabled complete protection at substantially lower doses when compared to vaccination with PspA + adjuvant. We also improved the storage stability of the hybrid vector to retain complete activity after 1 month at 4 °C using an approach in which hybrid vectors suspended in a microbial freeze drying buffer were desiccated. These results demonstrate the flexibility and robustness of the hybrid vector formulation, which has the potential to be a potent vaccine against S. pneumoniae.

RESUMEN

Most natural product biosynthetic gene clusters identified in bacterial genomic and metagenomic sequencing efforts are silent under laboratory growth conditions. Here, we describe a scalable biosynthetic gene cluster activation method wherein the gene clusters are disassembled at interoperonic regions in vitro using CRISPR/Cas9 and then reassembled with PCR-amplified, short DNAs, carrying synthetic promoters, using transformation assisted recombination (TAR) in yeast. This simple, cost-effective, and scalable method allows for the simultaneous generation of combinatorial libraries of refactored gene clusters, eliminating the need to understand the transcriptional hierarchy of the silent genes. In two test cases, this in vitro disassembly-TAR reassembly method was used to create collections of promoter-replaced gene clusters that were tested in parallel to identify versions that enabled secondary metabolite production. Activation of the atolypene ( ato) gene cluster led to the characterization of two unprecedented bacterial cyclic sesterterpenes, atolypene A (1) and B (2), which are moderately cytotoxic to human cancer cell lines. This streamlined in vitro disassembly- in vivo reassembly method offers a simplified approach for silent gene cluster refactoring that should facilitate the discovery of natural products from silent gene clusters cloned from either metagenomes or cultured bacteria.

Asunto(s)

RESUMEN

BACKGROUND AND OBJECTIVE: Cutaneous leishmaniasis (CL) is one of the most important diseases worldwide, with a different range of prevalence in endemic areas. Anthroponotic and zoonotic CL are two epidemiological forms of CL, in Iran. Although Ilam Province in the west of Iran is one of the main endemic areas of the disease, there is no inclusive study to determine the genetic variations of parasite in these areas. The objective of this study was to determine the genetic diversity of Leishmania species in Ilam Province, using mini-circle kDNA gene. MATERIALS AND METHODS: Direct smears were taken from skin lesions of 200 suspected cases of CL. Smears were stained, screened under light microscope. Polymerase chain reaction (PCR) was performed, using specific kinetoplast DNA primers. Data were analyzed, using the molecular bio-software. RESULTS: All the samples were positive by direct examination. PCR results showed all cases were positive for Leishmania major. Although all isolated cases belong to a different county of Ilam province, all were positive for L. major with intra-species genetic diversity, divided into four clades in the dendrogram. INTERPRETATION AND CONCLUSION: This variation can affect drug resistance and controlling strategies of parasite. It is possible that different species of sand flies and rodents are the vector and reservoir of parasite, respectively; however, further studies are needed to validate this.

RESUMEN

The Cytochrome P-4501B1 (CYP1B1) Leu432Val polymorphism has been previously shown to be associated with some types of cancer and affects CYP1B1-mediated metabolism of various infertility drugs. To establish the frequency of CYP1B1 Leu432Val polymorphism among women with a history of infertility drug use, we studied the genotypes of 147 patients with breast cancer with a history of infertility and 150 cancer-free, infertile women (control group) in Northern Iran. A polymerase chain reaction-based restriction fragment length polymorphism assay was used to detect GG (Val/Val), CG (Leu/Val), and CC (Leu/Leu) genotype frequencies, which did not vary significantly between the 2 patient groups (P = .847). We established for the first time that the incidence of CYP1B1 Leu432Val polymorphism is 46.6% among women with infertility history and breast cancer in Northern Iran. Finally, our results do not show any significant association between CYP1B1 Leu432Val polymorphism and breast cancer in infertile women in this region, who have also received infertility treatment.

RESUMEN

Yersiniabactin (Ybt) is a mixed nonribosomal peptide-polyketide natural product that binds a wide range of metals with the potential to impact processes requiring metal retrieval and removal. In this work, we substantially improved upon the heterologous production of Ybt and an associated anthranilate analog through systematic screening and optimization of culture medium components. Specifically, a Plackett-Burman design-of-experiments methodology was used to screen 22 components and to determine those contributing most to siderophore production. L-cysteine, L-serine, glucose, and casamino acids significantly contributed to the production of both compounds. Using this approach together with metabolic engineering of the base biosynthetic process, Ybt and the anthranilate analog titers were increased to 867 ± 121 mg/L and 16.6 ± 0.3 mg/L, respectively, an increase of ∼38 and ∼79-fold relative to production in M9 medium. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1193-1200, 2017.

Asunto(s)

RESUMEN

In this report, the heterologous production of salicylate (SA) is the basis for metabolic extension to salicylate 2-O-ß-d-glucoside (SAG), a natural product implicated in plant-based defense mechanisms. Production was optimized through a combination of metabolic engineering, gene expression variation, and co-culture design. When combined, SA and SAG production titers reached ~0.9g/L and ~2.5g/L, respectively. The SAG compound was then tested for anti-inflammatory properties relative to SA and acetylsalicylate (aspirin). Results indicate comparable activity between SAG and aspirin in reducing nitric oxide (NO) and reactive oxygen species (ROS) from macrophage cells while no discernable negative effects on cellular viability were observed.

Asunto(s)

RESUMEN

Biosynthesis of complex natural products like polyketides and nonribosomal peptides using Escherichia coli as a heterologous host provides an opportunity to access these molecules. The value in doing so stems from the fact that many compounds hold some therapeutic or other beneficial property and their original production hosts are intractable for a variety of reasons. In this work, metabolic engineering and induction variable optimization were used to increase production of the polyketide-nonribosomal peptide compound yersiniabactin, a siderophore that has been utilized to selectively remove metals from various solid and aqueous samples. Specifically, several precursor substrate support pathways were altered through gene expression and exogenous supplementation in order to boost production of the final compound. The gene expression induction process was also analyzed to identify the temperatures and inducer concentrations resulting in highest final production levels. When combined, yersiniabactin production was extended to ∼175 mg L-1 . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1412-1417, 2016.

Asunto(s)

RESUMEN

The type and potency of an immune response provoked during vaccination will determine ultimate success in disease prevention. The basis for this response will be the design and implementation of antigen presentation to the immune system. Whereas direct antigen administration will elicit some form of immunological response, a more sophisticated approach would couple the antigen of interest to a vector capable of broad delivery formats and designed for heightened response. New antigens associated with pneumococcal disease virulence were used to test the delivery and adjuvant capabilities of a hybrid biological-biomaterial vector consisting of a bacterial core electrostatically coated with a cationic polymer. The hybrid design provides (i) passive and active targeting of antigen-presenting cells, (ii) natural and multicomponent adjuvant properties, (iii) dual intracellular delivery mechanisms, and (iv) a simple formulation mechanism. In addition, the hybrid format enables device-specific, or in situ, antigen production and consolidation via localization within the bacterial component of the vector. This capability eliminates the need for dedicated antigen production and purification before vaccination efforts while leveraging the aforementioned features of the overall delivery device. We present the first disease-specific utilization of the vector toward pneumococcal disease highlighted by improved immune responses and protective capabilities when tested against traditional vaccine formulations and a range of clinically relevant Streptococcus pneumoniae strains. More broadly, the results point to similar levels of success with other diseases that would benefit from the production, delivery, and efficacy capabilities offered by the hybrid vector.

Asunto(s)

RESUMEN

E. coli has become a common host for the heterologous biosynthesis of natural products that demonstrate therapeutic value but suffer from access challenges posed by native production hosts. This review will highlight recent examples of heterologous products produced using E. coli. An emphasis will be placed on tools at the cellular and process levels to enable, improve, and alter production efforts. At the cellular scale, summaries of the process to enable heterologous biosynthesis will be supplemented with strategies (synthetic biology and metabolic engineering) to improve production levels. Process engineering strategies such as precursor-directed biosynthesis will also be highlighted in analog formation cases. In summary, the article will provide a recent overview of heterologous production efforts using E. coli and the relationship of the products produced to therapeutic applications.

Asunto(s)

RESUMEN

INTRODUCTION: Survival analysis for patients with end-stage renal disease and factors influencing their survival is crucial due to the increase in the number of these patients along with their high mortality rate. This study aimed to analyse the survival rate of patients in north of Iran undergoing hemodialysis and to assess factors influencing their survival. MATERIALS AND METHODS: A historical cohort study was conducted on 500 patients on maintenance hemodialysis in 3 hospitals of 2 cities (Sari and Babol) in Mazandaran province during a 6-year period from 2007 to 2013. The Cox regression analysis was used to assess the impact of sex, age, education, smoking habit, primary cause of kidney failure, living with family, cardiovascular diseases, weight, age at diagnosis, and age at initiating hemodialysis on survival of the patients. RESULTS: The median survival time for the 500 hemodialysis patients was 108 months. Death occurred in 174 patients (34.8%). History of smoking, age, being unemployed, being illiterate, and renal cyst, congenital diseases, and unspecified diseases as the cause of kidney failure were the associated factors with survival of the patients. The 1-, 2-, 3-, 5-, 10-, and 12-year survival for these patients was estimated to be 84%, 77%, 71%, 58%, 43%, and 33%, respectively. CONCLUSIONS: This study showed a high level of mortality and poor survival prognosis for patient undergoing maintenance hemodialysis. History of smoking, age, being unemployed, being illiterate, and renal cyst, congenital diseases, and unspecified conditions as the cause of kidney failure were the associated factors with survival of these patients.

Asunto(s)

RESUMEN

The production of the mixed nonribosomal peptide-polyketide natural product yersiniabactin (Ybt) has been established using E. coli as a heterologous host. In this study, precursor-directed biosynthesis was used to generate five new analogs of Ybt, demonstrating the flexibility of the heterologous system and the biosynthetic process in allowing compound diversity. A combination of biosynthetic and cellular engineering was then used to influence the production metrics of the resulting analogs. First, the cellular levels and activity of FadL, a hydrocarbon transport protein, were tested for subsequent influence upon exogenous precursor uptake and Ybt analog production with a positive correlation observed between FadL over-production and analog formation. Next, a Ybt biosynthetic editing enzyme was removed from the heterologous system which decreased native compound production but increased analog formation. A final series of experiments enhanced endogenous anthranilate towards complete pathway formation of the associated analog which showed a selective ability to bind gold.

Asunto(s)

RESUMEN

Yersiniabactin (Ybt) is a mixed nonribosomal peptide-polyketide natural product natively produced by the pathogen Yersinia pestis. The compound enables iron scavenging capabilities upon host infection and is biosynthesized by a nonribosomal peptide synthetase featuring a polyketide synthase module. This pathway has been engineered for expression and biosynthesis using Escherichia coli as a heterologous host. In the current work, the biosynthetic process for Ybt formation was improved through the incorporation of a dedicated step to eliminate the need for exogenous salicylate provision. When this improvement was made, the compound was tested in parallel applications that highlight the metal-chelating nature of the compound. In the first application, Ybt was assessed as a rust remover, demonstrating a capacity of ∼40% compared to a commercial removal agent and ∼20% relative to total removal capacity. The second application tested Ybt in removing copper from a variety of nonbiological and biological solution mixtures. Success across a variety of media indicates potential utility in diverse scenarios that include environmental and biomedical settings.

Asunto(s)

RESUMEN

Bactofection offers a gene delivery option particularly useful in the context of immune modulation. The bacterial host naturally attracts recognition and cellular uptake by antigen presenting cells (APCs) as the initial step in triggering an immune response. Moreover, depending on the bacterial vector, molecular biology tools are available to influence and/or overcome additional steps and barriers to effective antigen presentation. In this work, molecular engineering was applied using Escherichia coli as a bactofection vector. In particular, the bacteriophage ΦX174 lysis E (LyE) gene was designed for variable expression across strains containing different levels of lysteriolysin O (LLO). The objective was to generate a bacterial vector with improved attenuation and delivery characteristics. The resulting strains exhibited enhanced gene and protein release and inducible cellular death. In addition, the new vectors demonstrated improved gene delivery and cytotoxicity profiles to RAW264.7 macrophage APCs.

Asunto(s)

RESUMEN

Genetic vaccines offer a treatment opportunity based upon successful gene delivery to specific immune cell modulators. Driving the process is the vector chosen for gene cargo packaging and subsequent delivery to antigen-presenting cells (APCs) capable of triggering an immune cascade. As such, the delivery process must successfully navigate a series of requirements and obstacles associated with the chosen vector and target cell. In this work, we present the development and assessment of a hybrid gene delivery vector containing biological and biomaterial components. Each component was chosen to design and engineer gene delivery separately in a complimentary and fundamentally distinct fashion. A bacterial (Escherichia coli) inner core and a biomaterial [poly(beta-amino ester)]-coated outer surface allowed the simultaneous application of molecular biology and polymer chemistry to address barriers associated with APC gene delivery, which include cellular uptake and internalization, phagosomal escape, and intracellular cargo concentration. The approach combined and synergized normally disparate vector properties and tools, resulting in increased in vitro gene delivery beyond individual vector components or commercially available transfection agents. Furthermore, the hybrid device demonstrated a strong, efficient, and safe in vivo humoral immune response compared with traditional forms of antigen delivery. In summary, the flexibility, diversity, and potential of the hybrid design were developed and featured in this work as a platform for multivariate engineering at the vector and cellular scales for new applications in gene delivery immunotherapy.

Asunto(s)

RESUMEN

BACKGROUND: Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP. METHODS: Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes. RESULTS: The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum. CONCLUSION: C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.

RESUMEN

BACKGROUND: Metabolic syndrome (MS) is highly significant due to its association to type 2 diabetes and cardiovascular diseases. The purpose of this study was to compare the prevalence of MS according to the report of the Iranian National Committee of Obesity criteria (INCO) versus Adult Treatment Panel III (ATPIII) in Babol, North of Iran. METHODS: Data obtained based on criteria ATP III from the Babol Lipid and Glucose Study (from July 2004 to September 2005) and were compared with the new INCO criteria 2010. The data were collected and analyzed. RESULTS: In total, 933 adult males and females were evaluated. According to ATP III criteria, the overall prevalence of metabolic syndrome was 23.7% (95% confidence interval: 21%-26.4%); 28.4% and 9.4% were females and males, respectively; however, the prevalence was 20.5% (95% confidence interval: 17.9%-23.1%) according to the INCO criteria, 22.5% and 15.7% were females and males, respectively. CONCLUSION: The new INCO criteria for the metabolic syndrome proclaimed by the Iranian Committee of Obesity estimated a lower prevalence of syndrome in comparison with ATP III criteria in Babol.

RESUMEN

BACKGROUND: Patients with pelvic mass are the most referred patients to gynecologist. The aim of this study was to evaluate the ability of three malignancy risk indices (RMI 1, RMI 2 and RMI 3) and CA-125 to discriminate a benign from a malignant pelvic mass in our region (North of Iran). METHODS: This retrospective study was performed on 182 women with pelvic masses referred to Yahyanejad Hospital from 2007 to 2009. Ultrasound scans were scored as one point for each of the following characteristics: multilocular cyst, solid areas, intra-abdominal metastases, ascites, and bilateral lesions. For each patient a total ultrasound score (U) was calculated. The difference of the three RMI was based on the allocation of the U and M scores. The sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) of level of serum CA-125, the RMI 1, 2 and 3 were compared. RESULTS: Mean age of the patients was 39.9 ± 9.3 years. Most of them were premenopausal (161 women or 88.4%). A significant linear trend for malignancy was found by increasing age, ultrasound score, and serum CA-125. The best performance of CA125 was at a cut-off 88 U/ml, with a sensitivity of 88%, a specificity of 97%, a positive predictive value of 84%, and a negative predictive value of 99%. RMI 1 and 3 at the optimal cut off point of 265 and RMI2 at the optimal cut off point of 355, had a sensitivity of 91%, specificity of 96%, a positive predictive value of 78%, and a negative predictive value of 99%. CONCLUSION: In our population we found that there is no statistically significant difference in the performance of three malignancy risk indices (RMI 1, RMI 2, and RMI 3) and CA125 in differentiating between benign and malignant pelvic masses.