Improving AOX1 promoter efficiency by overexpression of Mit1 transcription factor.

BACKGROUND: Reprogramming in transcriptional regulation provides an effective tool for adjusting cellular metabolic activities. The strong methanol-inducible alcohol oxidase-1 promoter (pAOX1) is commonly used for heterologous gene expression in the yeast Pichia pastoris. Here, we present a novel Pichia pastoris strain engineered to co-express methanol-induced transcription factor 1 (Mit1) and the target protein. Mit1 upregulates pAOX1 in response to methanol. METHODS AND RESULTS: Two model proteins (VEGF and eGFP) have been used as the target proteins under the control of pAOX1. The sequence of Mit1 had obtained from the yeast genome and likewise cloned under the control of pAOX1. The results indicated a 1.9 and 2.2 fold increase in the detected VEGF and eGFP, respectively, when co-expressed with Mit1. Furthermore, the double-recombinant cells, containing Mit-1 and eGFP, produced 1.3 fold more eGFP when the methanol feeding concentration was doubled. The real-time PCR indicated a slight increase in the Mit1 expression, probably due to the negative regulatory feedback loop that exists for the intrinsic yeast Mit1. Overexpression of Mit1 also led to duplication of AOX1 enzyme activity, which may enhance the yeast cells' capacity for methanol detoxification. CONCLUSION: Overexpression of Mit1 could be considered a promising strategy for upregulation of target recombinant proteins in Pichia pastoris. Intracellular overexpression of Mit1 upregulates the heterologous target gene (eGFP) production, which is expressed under the control of pAOX1.

Does laparoscopic hysterectomy + bilateral salpingectomy decrease the ovarian reserve more than total abdominal hysterectomy? A cohort study, measuring anti-Müllerian hormone before and after surgery.

BACKGROUND: Decreased ovarian function and reserve is one of the complications of hysterectomy. In this study, we aimed to compare anti-müllerian hormone (AMH) levels between total abdominal hysterectomy (TAH), and total laparoscopic hysterectomy (TLH). METHODS: In this prospective cohort study, serum levels of AMH were compared between the groups undergoing TAH + bilateral salpingectiomy and TLH, in 66 patients (33 in each group) who referred to the hospitals of Shiraz University of Medical Sciences for hysterectomy during one years of work. The collected information included age, weight, gravidity, parity, regularity of menstrual cycle, uterine weight, blood loss during surgery, and serum levels of AMH before and 6 months after surgery, compared between groups. RESULTS: Most patients (88% in TAH and 73% in TLH group) aged 40-50 years. Mean age, weight, parity of patients was similar in both groups, while blood loss was significantly less in TLH group (P < 0.01). Median (IQR) of pre-surgical AMH values were 0.40 (0.55) ng/ml in the TLH group and 0.92 (1.23) ng/ml in the TAH group (P = 0.12) that decreased to 0.29 (0.44) ng/ml in the TLH group and 0.15 (0.31) ng/ml in the TAH group (P = 0.02). Also Median (IQR) of the difference between pre and post-surgical AMH values were 0.12 (0.31) and 0.58 (1.17) in TLH and TAH group, respectively (P = 0.003). CONCLUSION: The serum levels of AMH decreased significantly after both methods of hysterectomy (laparoscopy and laparotomy), while this decrease was greater in TAH group that shows.

Zoonotic parasitic organisms on vegetables: Impact of production system characteristics on presence, prevalence on vegetables in northwestern Iran and washing methods for removal.

Fresh vegetables are essential components of a healthy and nutritious diet, but if consumed raw without proper washing and/or disinfection, can be important agents of transmission of enteric pathogens. This study aimed to determine the prevalence of zoonotic parasites on vegetables freshly harvested and "ready to eat" vegetables from greengrocers and markets in northwestern Iran. In addition, the effect of cropping system and season on contamination levels were assessed as well as the efficacy of washing procedures to remove parasites from the vegetables. A total of 2757 samples composed of field (n = 1, 600) and "ready to eat" (n = 1157) vegetables were analyzed. Vegetables included leek, parsley, basil, coriander, savory, mint, lettuce, cabbage, radish, dill, spinach, mushroom, carrot, tomato, cucumber and pumpkin. Normal physiological saline washings from 200 g samples were processed using standard parasitological techniques and examined microscopically. A total of 53.14% of vegetable samples obtained from different fields and 18.23% of "ready to eat" vegetables purchased from greengrocers and markets were contaminated with different parasitic organisms including; Entamoeba coli cysts, Giardia intestinalis cysts, Cryptosporidium parvum oocysts, Fasciola hepatica eggs, Dicrocoelium dendriticum eggs, Taenia spp. eggs, Hymenolepis nana eggs, Ancylostoma spp. eggs, Toxocara cati eggs, Toxocara canis eggs, Strongyloides stercoralis larvae, and Ascaris lumbricoides eggs. In both field and "ready to eat" vegetables, the highest parasitic contamination was observed in lettuce with a rate of 91.1% and 55.44%, respectively. The most common parasitic organism was Fasciola hepatica. A seasonal difference in contamination with parasitic organisms was found for field and "ready to eat" vegetables (P < 0.05). There was a significant difference in the recovery of parasitic organisms depending on the washing method with water and dishwashing liquid being the least effective. Proper washing of vegetables is imperative for a healthy diet as the results of this study showed the presence of zoonotic parasites from field and ready to eat vegetables in Iran.

Assessing the risk factors before pregnancy of preterm births in Iran: a population-based case-control study.

BACKGROUND: Preterm birth is a major cause of prenatal and postnatal mortality particularly in developing countries. This study investigated the maternal risk factors associated with the risk of preterm birth. METHODS: A population-based case-control study was conducted in several provinces of Iran on 2463 mothers referred to health care centers. Appropriate descriptive and analytical statistical methods were used to evaluate the association between maternal risk factors and the risk of preterm birth. All tests were two-sided, and P values < 0.05 were considered to be statistically significant. RESULTS: The mean gestational age was 31.5 ± 4.03 vs. 38.8 ± 1.06 weeks in the case and control groups, respectively. Multivariate regression analysis showed a statistically significant association between preterm birth and mother's age and ethnicity. Women of Balooch ethnicity and age ≥ 35 years were significantly more likely to develop preterm birth (OR: 1.64; 95% CI: 1.01--2.44 and OR: 9.72; 95% CI: 3.07-30.78, respectively). However, no statistically significant association was observed between preterm birth and mother's place of residence, level of education, past history of cesarean section, and BMI. CONCLUSION: Despite technological advances in the health care system, preterm birth still remains a major concern for health officials. Providing appropriate perinatal health care services as well as raising the awareness of pregnant women, especially for high-risk groups, can reduce the proportion of preventable preterm births.

Effect of age, sex, and season on the prevalence of Linguatula serrata infestation in mesenteric lymph nodes of goats slaughtered in Tabriz, Iran.

This research was conducted to determine the prevalence and intensity of infection with nymphs of Linguatula serrata in mesenteric lymph nodes of the goats from northwest of Iran. Moreover, the intensity of infection of mesenteric lymph nodes was compared based on gross appearance and the morphological changes in these nodes. In addition, the effects of age, sex, and season on the prevalence of Linguatula serrata were assessed. In this study, mesenteric lymph nodes (n = 16,284) were randomly collected from 2132 goats slaughtered in the abattoir of Tabriz, Iran, from September 2013 to September 2017. The samples were categorized based on their gross appearance (color and consistency) and then were cut into small pieces and immersed in normal saline (0.9% NaCl) solution and left for 5 to 6 h to allow nymphs to come out from the tissue. The results showed that 1241 out of total 2132 (58. 2%) goats and 9282 out of total 16,284 mesenteric lymph nodes (57%) were infested with the nymphal stage of Linguatula serrata. It was also revealed that the infestation rate was age-dependent: as the goats grow older, the infestation increases. Further, the prevalence of Linguatula serrata nymphs in the mesenteric lymph nodes in various seasons was not significant (P > 0.05). The prevalence of infestation rate in female goats was significantly higher than of male goats (P < 0.05). Besides, the infestation rate in the black-colored lymph nodes (75.88%) was significantly (P < 0.05) higher than those of hemorrhagic nodes (54.94%) and normal-colored nodes (22.65%). Moreover, the infestation rate of nymphs in the soft lymph nodes (83.91%) was significantly (P < 0.05) higher than those in normal (21.85%) and hard (32.43%) lymph nodes. Given the fact that the Linguatula serrata is a zoonotic parasite; thus, the inspection process should be meticulously done in an abattoir, especially in areas where residents consume raw or under-cooked liver and/or visceral organs of herbivores.

Genetic study of the BRAF gene reveals new variants and high frequency of the V600E mutation among Iranian ameloblastoma patients.

BACKGROUND: Ameloblastoma is a benign, slow-growing and locally invasive tumor. It is one of the most prevalent odontogenic tumors, with an incidence rate of 1% of all oral tumors and approximately 18% of odontogenic tumors. A group of genes have been investigated in patients with ameloblastoma. The BRAF V600E mutation has been implicated as the most common mutation in ameloblastoma. The presence or absence of this mutation has been associated with several clinicopathological properties, including location, age at diagnosis, histology, and prognosis. Although some populations have been investigated so far, little data are available on the Iranian population. The current research was launched to study the BRAF V600E mutation among a cohort of Iranian patients with ameloblastoma. METHODS: In this clinicopathological and molecular biology study, a total of 19 formalin-fixed, paraffin-embedded tissues were studied. DNA extraction was performed, followed by PCR-sequencing of exons 10 and 15 of the BRAF gene to identify mutations. In silico analysis was performed for the identified variants. Results were analyzed by T test, Chi-square, and Fisher's exact test. RESULTS: Totally, 12 of 19 samples (63%) harbored the p. V600E hotspot mutation. In addition, we identified several variants, two of which were novel. The c.1769T>G (p. V590G) and c.1751C>T (p.L584F) as the novel variants showed a possible damaging effect by in silico analysis. No variant was found within exon 10. CONCLUSIONS: Our study confirms the role of BRAF mutations in ameloblastoma in the Iranian patients studied.

Developing a methodology for real-time trading of water withdrawal and waste load discharge permits in rivers.

In this paper, a new methodology is proposed for the real-time trading of water withdrawal and waste load discharge permits in agricultural areas along the rivers. Total Dissolved Solids (TDS) is chosen as an indicator of river water quality and the TDS load that agricultural water users discharge to the river are controlled by storing a part of return flows in some evaporation ponds. Available surface water withdrawal and waste load discharge permits are determined using a non-linear multi-objective optimization model. Total available permits are then fairly reallocated among agricultural water users, proportional to their arable lands. Water users can trade their water withdrawal and waste load discharge permits simultaneously, in a bilateral, step by step framework, which takes advantage of differences in their water use efficiencies and agricultural return flow rates. A trade that would take place at each time step results in either more benefit or less diverted return flow. The Nucleolus cooperative game is used to redistribute the benefits generated through trades in different time steps. The proposed methodology is applied to PayePol region in the Karkheh River catchment, southwest Iran. Predicting that 1922.7 Million Cubic Meters (MCM) of annual flow is available to agricultural lands at the beginning of the cultivation year, the real-time optimization model estimates the total annual benefit to reach 46.07 million US Dollars (USD), which requires 6.31 MCM of return flow to be diverted to the evaporation ponds. Fair reallocation of the permits, changes these values to 35.38 million USD and 13.69 MCM, respectively. Results illustrate the effectiveness of the proposed methodology in the real-time water and waste load allocation and simultaneous trading of permits.

Psychometric Properties of the Persian Version of the God Locus of Health Control (GLHC): A Study on Muslim Pilgrims.

God Locus of Health Control (GHLC) scale is an instrument for assessing to what extent one believes that God controls his/her health or disease. The aim of this study was to describe the cultural adaptation of this scale and to investigate the psychometric properties of Persian version. At first, the scale was forward-translated to Persian, and then, backward-translation was conducted. Finally, the synthesized back-translated version was prepared and compared to the draft Persian-language version of the GLHC. In the next step, in order to test cognitive debriefing, the Persian-language version of the questionnaire was completed by 20 pilgrims. Final version of Persian GHLC scale was answered by 600 pilgrims attending religious holy places. Mean age of respondents was 33.15 (±11.04) years. There was a positive correlation between six items of GLHC scale and inter-item correlation coefficients. The Cronbach's alpha coefficient was 0.84. Exploratory factor analysis showed that the G LHC scale had a one-factor structure. The factor loadings for all items ranged from 0.57 to 0.77. Residual correlations of all items were between -0.2 and +0.2. The people with lower income had significantly higher score of GLHC scale. GLHC scale score had a significant positive correlation with DUREL subscale and MHLC Chance Externality subscale score. There was no a significant correlation between GLHC scale score and Multidimensional Health Locus of Control (MHLC) Internality and Powerful subscales scores. The results of the present study showed that the Persian version of GHLC scale has acceptable factorial validity and internal consistency reliability.

Effects of narrow-base walking and dual tasking on gait spatiotemporal characteristics in anterior cruciate ligament-injured adults compared to healthy adults.

PURPOSE: The present experiment was conducted to examine the hypothesis that challenging control through narrow-base walking and/or dual tasking affects ACL-injured adults more than healthy control adults. METHODS: Twenty male ACL-injured adults and twenty healthy male adults walked on a treadmill at a comfortable speed under two base-of-support conditions, normal-base versus narrow-base, with and without a cognitive task. Gait patterns were assessed using mean and variability of step length and mean and variability of step velocity. Cognitive performance was assessed using the number of correct counts in a backward counting task. RESULTS: Narrow-base walking resulted in a larger decrease in step length and a more pronounced increase in variability of step length and of step velocity in ACL-injured adults than in healthy adults. For most of the gait parameters and for backward counting performance, the dual-tasking effect was similar between the two groups. CONCLUSIONS: ACL-injured adults adopt a more conservative and more unstable gait pattern during narrow-base walking. This can be largely explained by deficits of postural control in ACL-injured adults, which impairs gait under more balance-demanding conditions. The observation that the dual-tasking effect did not differ between the groups may be explained by the fact that walking is an automatic process that involves minimal use of attentional resources, even after ACL injury. Clinicians should consider the need to include aspects of terrain complexity, such as walking on a narrow walkway, in gait assessment and training of patients with ACL injury. LEVEL OF EVIDENCE: III.

Universal depression screening, diagnosis, management, and outcomes at a student-run free clinic.

OBJECTIVE: Student-run free clinics (SRFCs) are now present at most medical schools. Reports regarding SRFCs have focused on the infrastructure of established clinics, characteristics of the patient populations served, and their contribution to patient care. Few studies discuss their role in preventive medicine and even fewer discuss mental health care. This study examined the outcomes of a medical student-run universal depression screening, diagnosis, and management program at two SRFC sites. METHODS: Medical students implemented a universal depression screening, diagnosis, and management program within the electronic health record during routine adult primary care visits utilizing the Patient Health Questionnaire-2 (PHQ-2) as an initial screening tool, with a protocol to administer the Patient Health Questionnaire-9 (PHQ-9) if the PHQ-2 score was ≥3. This is a retrospective medical record review of visits from August 13, 2013, through February 13, 2014, to assess this program. RESULTS: Overall, 95.8 % (206/215) of the patients received either the PHQ-2 or the PHQ-9. Among the 174 patients without a previous diagnosis of depression, 166 were screened (95.4 %), of which 33 (19.9 %) had a positive PHQ-2 score of ≥3; 30 (of 33; 90.9 %) appropriately received a PHQ-9. Nineteen (of 166 screened; 11.4 %) previously undiagnosed patients were confirmed to have depression. Fourteen patients had two or more PHQ-9 tests at least 4 weeks apart and eight (57.1 %) had a clinically significant improvement, defined as PHQ-9 score decrease of ≥5. The prevalence of depression diagnosed prior to the implementation of this program in this cohort was 19.1 % (41/215) and after was 27.9 % (60/215). CONCLUSIONS: This study demonstrated that medical students with faculty supervision can successfully implement a universal depression screening, diagnosis, and management program at multiple SRFC sites, identify previously undiagnosed depression, and work with interdisciplinary support services to provide treatment options, leading to a clinically significant improvement in depression severity.

River water quality management considering agricultural return flows: application of a nonlinear two-stage stochastic fuzzy programming.

In this paper, a new fuzzy methodology is developed to optimize water and waste load allocation (WWLA) in rivers under uncertainty. An interactive two-stage stochastic fuzzy programming (ITSFP) method is utilized to handle parameter uncertainties, which are expressed as fuzzy boundary intervals. An iterative linear programming (ILP) is also used for solving the nonlinear optimization model. To accurately consider the impacts of the water and waste load allocation strategies on the river water quality, a calibrated QUAL2Kw model is linked with the WWLA optimization model. The soil, water, atmosphere, and plant (SWAP) simulation model is utilized to determine the quantity and quality of each agricultural return flow. To control pollution loads of agricultural networks, it is assumed that a part of each agricultural return flow can be diverted to an evaporation pond and also another part of it can be stored in a detention pond. In detention ponds, contaminated water is exposed to solar radiation for disinfecting pathogens. Results of applying the proposed methodology to the Dez River system in the southwestern region of Iran illustrate its effectiveness and applicability for water and waste load allocation in rivers. In the planning phase, this methodology can be used for estimating the capacities of return flow diversion system and evaporation and detention ponds.

Water and waste load allocation in rivers with emphasis on agricultural return flows: application of fractional factorial analysis.

In this paper, a new methodology is developed to handle parameter and input uncertainties in water and waste load allocation (WWLA) in rivers by using factorial interval optimization and the Soil, Water, Atmosphere, and Plant (SWAP) simulation model. A fractional factorial analysis is utilized to provide detailed effects of uncertain parameters and their interaction on the optimization model outputs. The number of required optimizations in a fractional factorial analysis can be much less than a complete sensitivity analysis. The most important uncertain inputs and parameters can be also selected using a fractional factorial analysis. The uncertainty of the selected inputs and parameters should be incorporated real time water and waste load allocation. The proposed methodology utilizes the SWAP simulation model to estimate the quantity and quality of each agricultural return flow based on the allocated water quantity and quality. In order to control the pollution loads of agricultural dischargers, it is assumed that a part of their return flows can be diverted to evaporation ponds. Results of applying the methodology to the Dez River system in the southwestern part of Iran show its effectiveness and applicability for simultaneous water and waste load allocation in rivers. It is shown that in our case study, the number of required optimizations in the fractional factorial analysis can be reduced from 64 to 16. Analysis of the interactive effects of uncertainties indicates that in a low flow condition, the upstream water quality would have a significant effect on the total benefit of the system.

Isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings.

BACKGROUND: Invasive fungal infections cause considerable morbidity and mortality in immunocompromised hosts. Pigeon droppings could especially be a potential carrier in the spread of pathogenic yeasts and mold fungi into the environment. The objective of this study was to isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings. MATERIALS AND METHODS: One hundred twenty samples of pigeon droppings were suspended 1:10 in saline solution and then cultured. Identification of C. neoformans was performed on bird seed agar, presence of a capsule on India ink preparation, urease production on urea agar medium and RapID yeast plus system. The identification of candida species was based on micro-morphological analysis on corn meal-Tween 80 agar, RapID yeast plus system and growth in CHROMagar candida. The identification of other fungi was based on macromorphologic, microscopic, biochemical and physiological characteristics. RESULTS: The highest frequency of yeasts and mold fungi were observed in Candida albicans 6.6% and Penicillium spp. 25%. The frequency rate of C. neoformans isolation was 2.5%. CONCLUSION: Several types of fungi are present in pigeon droppings that can spread in environment and transmit to children and elderly as well as immunocompromised patients who are at increased risk of contracting opportunistic diseases.

Polyamidoamine with a hyper-branched structure grafted on modified magnetic graphene oxide for the trace separation of diclofenac and acetaminophen followed by high-performance liquid chromatography determination.

Different generations of polyamidoamine dendrimers were synthesized on a focal core of magnetic graphene oxide modified with 3-aminopropyltriethoxysilane. After the characterization of synthesized dendrimers, its second generation was employed as a suitable sorbent for simultaneous separation/preconcentration of diclofenac and acetaminophen by a dispersive magnetic solid phase microextraction. The extracted analytes were then quantified by high-performance liquid chromatography with ultraviolet detection. Under optimized conditions, the limits of detection were 0.3 µg/L for diclofenac and 0.1 µg/L for acetaminophen. The intra-day relative standard deviations at 50 µg L-1 levels were 1.8% for diclofenac and 2.1% for acetaminophen, while the inter-day relative standard deviations were 3.6% and 4.5% for diclofenac and acetaminophen, respectively. The calibration graphs were linear in ranges of 1.0-500.0 µg/L and 0.5-600.0 µg/L for diclofenac and acetaminophen, respectively, with good coefficients of determination (r2 > 0.998). The method was successfully applied to the determination of diclofenac and acetaminophen in water, milk, and biological samples.

Clinical EEG slowing induced by electroconvulsive therapy is better described by increased frontal aperiodic activity.

Electroconvulsive therapy (ECT) is one of the most efficacious interventions for treatment-resistant depression. Despite its efficacy, ECT's neural mechanism of action remains unknown. Although ECT has been associated with "slowing" in the electroencephalogram (EEG), how this change relates to clinical improvement is unresolved. Until now, increases in slow-frequency power have been assumed to indicate increases in slow oscillations, without considering the contribution of aperiodic activity, a process with a different physiological mechanism. In this exploratory study of nine MDD patients, we show that aperiodic activity, indexed by the aperiodic exponent, increases with ECT treatment. This increase better explains EEG "slowing" when compared to power in oscillatory peaks in the delta (1-3 Hz) range and is correlated to clinical improvement. In accordance with computational models of excitation-inhibition balance, these increases in aperiodic exponent are linked to increasing levels of inhibitory activity, suggesting that ECT might ameliorate depressive symptoms by restoring healthy levels of inhibition in frontal cortices.

A comparison of interactive immersive virtual reality and still nature pictures as distraction-based analgesia in burn wound care.

PURPOSE: Non-pharmacologic adjuncts to opioid analgesics for burn wound debridement enhance safety and cost effectiveness in care. The current study explored the feasibility of using a custom portable water-friendly immersive VR hardware during burn debridement in adults, and tested whether interactive VR would reduce pain more effectively than nature stimuli viewed in the same VR goggles. METHODS: Forty-eight patients with severe burn injuries (44 adults and 4 children) had their burn injuries debrided and dressed in a wet wound care environment on Study Day 1, and 13 also participated in Study Day 2. INTERVENTION: The study used a within-subject design to test two hypotheses (one hypothesis per study day) with the condition order randomized. On Study Day 1, each individual (n = 44 participants) spent 5 min of wound care in an interactive immersive VR environment designed for burn care, and 5 min looking at still nature photos and sounds of nature in the same VR goggles. On Study Day 2 (n = 12 adult participants and one adolescent from Day 1), each participant spent 5 min of burn wound care with no distraction and 5 min of wound care in VR, using a new water-friendly VR system. On both days, during a post-wound care assessment, participants rated and compared the pain they had experienced in each condition. OUTCOME MEASURES ON STUDY DAYS 1 AND 2: Worst pain during burn wound care was the primary dependent variable. Secondary measures were ratings of time spent thinking about pain during wound care, pain unpleasantness, and positive affect during wound care. RESULTS: On Study Day 1, no significant differences in worst pain ratings during wound care were found between the computer-generated world (Mean = 71.06, SD = 26.86) vs. Nature pictures conditions (Mean = 68.19, SD = 29.26; t < 1, NS). On secondary measures, positive affect (fun) was higher, and realism was lower during computer-generated VR. No significant differences in pain unpleasantness or "presence in VR" between the two conditions were found, however. VR VS. NO VR. (STUDY DAY 2): Participants reported significantly less worst pain when distracted with adjunctive computer generated VR than during standard wound care without distraction (Mean = 54.23, SD = 26.13 vs 63.85, SD = 31.50, t(11) = 1.91, p < .05, SD = 17.38). In addition, on Study Day 2, "time spent thinking about pain during wound care" was significantly less during the VR condition, and positive affect was significantly greater during VR, compared to the No VR condition. CONCLUSION: The current study is innovative in that it is the first to show the feasibility of using a custom portable water-friendly immersive VR hardware during burn debridement in adults. However, contrary to predictions, interactive VR did not reduce pain more effectively than nature stimuli viewed in the same VR goggles.