The stochastic frontier analysis technique in measuring the technical and economic efficiency of hospital diagnostic laboratories: a case study in Iran.

An inefficient health system wastes scarce resources even if it makes considerable gains in accountability and equity. Such a system is expected to perform better. Therefore, it is vital to examine the current performance of health systems and their constituents and assess how to reach their maximum potential. This study aimed to evaluate the technical and economic efficiency of medical diagnostic laboratories in hospitals affiliated with Urmia University of Medical Sciences (UUMS) in 2016. In this descriptive-analytical study, data from diagnostic laboratories of the hospitals of UUMS have been inputted into Frontier4.1 software after taking the log of variables. Then, the technical and economic efficiency of the laboratories were obtained by estimating the production and cost function using the stochastic frontier analysis method, assuming input minimization for 2016. The mean technical and economic efficiency score of the diagnostic laboratories were determined to be 93.1% and 51.9%, respectively. These laboratories need to reduce their inputs and costs in order to achieve full efficiency without changing the amount of their output. Although the average economic efficiency of the diagnostic laboratories of the studied hospitals was high, there is still an increase in the efficiency of these units, given the cost of inputs at the time of allocating resources. In addition, it is possible to improve the technical efficiency of the clinical laboratories of hospitals affiliated with UUMS by 48.1% by applying the same level of inputs and without increasing the costs.

Sero-epidemiology of Hydatidosis Among General Population of Jolfa County, Northwestern Iran Using IHA, ELISA and Western Blot (2017-2018).

BACKGROUND: Human hydatidosis is mostly a latent and neglected disease with known endemicity in Iran. AIMS: Due to the importance of this infection in the country and its latent nature, we aimed to evaluate the serological status of hydatid cyst in northwestern Iran. OBJECTIVES: Herein, we evaluated the serological status of hydatid cyst in urban and rural inhabitants of Jolfa county, northwestern Iran during 2017-2018. METHODS: In total, 1296 blood samples were obtained from human individuals and the presence of anti-E. granulosus antibodies was investigated using IHA, ELISA and WB. RESULTS: Based on results, 25 IHA positive person were detected in the examined population, however ELISA test showed 14 of 25 IHA positive patients as negative. Also, 269 IHA negative fellows were shown as negative by ELISA. WB analysis of sera from 25 IHA positive subjects revealed consistent results with the ELISA test, and the most reactive SHCF Ag was a 37 KDa protein. The age-standardized seroprevalence of hydatidosis among Jolfa's general population was 1.12% with 95%CI: 1.02-1.20. Moreover, there existed a significant association between keeping/- contact with dogs (P = 0.022) as well as vegetable consumption (P < 0.001) with ELISA positive test results. CONCLUSION: Along with such serological evidence in this region, we highly suggest physical examination and applying imaging techniques for suspected cases in the area for a better understanding of CE.

Plasmapheresis, Anti-ACE2 and Anti-FcγRII Monoclonal Antibodies: A Possible Treatment for Severe Cases of COVID-19.

In March 2020, the WHO declared the COVID-19 disease as a pandemic disease. There have been studies on the COVID-19 to find a certain treatment, but yet, there is no certain cure. In this article, we present a possible way to treat severe cases of COVID-19. Based on the previous studies, there are similarities between the spike antigens of SARS-CoV and SARS-CoV-2 viruses. It is expected that these similarities (structural and affinity to the receptor of ACE2) can lead to the same pathophysiological activity of the virus by the use of ACE2 and FcγRII (the antibody-dependent enhancement mechanism). Therefore, we propose a way of washing out (by plasmapheresis) the possible antibodies against the spike protein of the virus out of patients' plasma to stop the antibody-dependent enhancement (ADE)-mediated infection of the immune system cells at the first phase of the treatment and simultaneous use of the anti-ACE2 with anti-FcγRII monoclonal antibodies at the second phase. We propose these procedures for the patients that have no significant response for typical anti-viral, ARDS and conservative therapies, and the disease persists or progresses despite sufficient therapies.

Hsp90 Inhibitor; NVP-AUY922 in Combination with Doxorubicin Induces Apoptosis and Downregulates VEGF in MCF-7 Breast Cancer Cell Line.

OBJECTIVE: Breast cancer is one of the most prevalent malignancies and leading causes of females' mortality worldwide. Because of resistance to various treatment options, new treatments based on molecular targeting has introduced as noticeable strategies in cancer treatment. In this regard, heat shock protein 90 (Hsp90) inhibitors are proposed as effective anticancer drugs. The goal of the study was to utilize a combination of the doxorubicin (DOX) and NVP-AUY 922 on the MCF-7 breast cancer model to investigate the possible cytotoxic mechanisms. METHODS: MCF-7 breast cancer cell line was prepared and treated with various concentrations of DOX and NVP-AUY922 in single-drug treatments. We investigated the growth-inhibitory pattern by MTT assay after continuous exposure to NVP-AUY922 and DOX in order to determine dose-response. Then the combinatorial effects were evaluated in concentrations of 0.5 × IC50, 0.2 × IC50, 1 × IC50 and, 2 × IC50 of each drugs. Based on MTT results of double combinations, low effective doses were selected for Real-time PCR [caspase3 and vascular endothelial growth factor(VEGF)] and caspase 3 enzyme activity. RESULTS: A dose-dependent inhibitory effects were presented with increasing the doses of both drugs in single treatments. The upregulation of caspase 3 and downregulation of VEGF mRNA were observed in double combinations of NVP-AUY922 and DOX versus single treatments. Also, in these combinations in low doses of examined drugs (0.5 × IC50, 0.2 × IC50), higher caspase 3 activity were presented in comparison to single treatments (p<0.05). CONCLUSIONS: Our findings indicate an effective action of NVP-AUY922 in combined with DOX in this cell line. These results can predict the treatment outcome in this model.

The economic efficiency of clinical laboratories in public hospitals: A case study in Iran.

INTRODUCTION: Clinical laboratories are identified as one of the most important and expensive units of the health system. Therefore, it is essential to pay attention to these units' cost efficiency. This study purpose was to evaluate the economic efficiency of hospitals' laboratory units affiliated to Urmia University of Medical Sciences (UMSU), in order to assess their performance. METHODS: This research was a descriptive-analytic study that was accomplished in 2017. The statistical population of the study included all of the hospitals' clinical laboratories affiliated to UMSU. Moreover, DEA method and Deap2.1 software were used to analyze data. In this study, technical and allocative efficiencies of the studied laboratory units were also calculated in addition to the determination of the economic efficiency of the laboratories. RESULTS: The average economic efficiency of clinical laboratories calculated by DEA in 2017 was 0.676. This value was lower than the allocative and technical efficiency scores, which indicates that these units could attain full efficiency by reducing their costs without having any effect on output values. Moreover, about 14 percent of the clinical laboratory units were economically efficient. In addition, it is noteworthy to state that, from total of university hospital laboratories, only three hospitals had no economic excess or deficiency values of inputs. CONCLUSION: Considering that 76% of laboratory units have not been economically efficient, it is necessary for the laboratory managers to consider optimum allocating of resources, with respect to the cost of laboratory equipment and inputs in order to increase their units' economic efficiency.

Expression and purification of biologically active recombinant rabbit monocyte chemoattractant protein1 in Escherichia coli.

Monocyte chemoattractant protein 1 (MCP1) with recruiting monocytes is an important factor at the beginning of inflammatory disorders such as atherosclerosis which seems its blocking preclude this process and help improvement of related diseases. To perform clinical research in this field, MCP1 protein is required but firstly, animal studies should be done. As the rabbit is a suitable model for many inflammatory disorders, and Escherichia coli BL21(DE3) (BL21) cell is a high-efficiency host for protein expression, we decided to produce recombinant rabbit MCP1 (rRMCP1) in BL21/pET28a system. After codon usage, a construct containing RMCP1 sequence was synthesized, cloned into the pET28a plasmid, and overexpressed in BL21 cells. Followed that, with changing expression condition such as cell concentration before the induction, time period, temperature, shaking rate and inducer concentration (IPTG), rRMCP1 expression was optimized, and purified by Ni-NTA. The biological activity of the expressed protein was verified using monocyte migration assay. Using this expression system, nearly 28 mg/mL rRMCP1 was produced at 26°C/180 rpm for 24 h in LB broth medium with 1 mM IPTG. Therefore, we were succeeded to express the intermediate level of rRMCP1 with this method. This amount of protein is sufficient for biological researches in the laboratory.

The Increase in Protein and Plasmid Yields of E. coli with Optimized Concentration of Ampicillin as Selection Marker.

Background:Escherichia coli is still the common host for ing and heterologous protein expression. Various strategies have been employed to increase protein expression in E. coli, but, it seems that external factors such as selection marker concentration can drastically affect the yield of protein and plasmid. Objectives: Alterations of protein expression and plasmid yields of E. coli in different concentrations of ampicillin, as selection marker, will be determined. In order to improve heterologous expression, the system will be redesigned and optimized. Materials and Methods: The expression cassette of codon optimized EGFP for E. coli was synthesized in pUC57. The pUC57-GFP was transformed into E. coli Top10F'. The expression of GFP was verified by SDS-PAGE and flow cytometry after induction by IPTG (0.5 mM) and incubation with 0, 100, 200 and 300 µg.mL-1 ampicillin. Plasmid copy numbers of samples were determined by Real-Time PCR on AMP gene using regression line of diluted standard curve. Results: GFP expressing clones formed fair green colonies on LB agar supplemented with 0.5 mM IPTG and showed fluorescence in FL1 filter of flow cytometry and an extra protein band on SDS-PAGE gel. The fluorescent intensity of GFP in 0, 100, 200 and 300 µg.mL-1 ampicillin in medium were 549.83, 549.78, 1443.52, 684.87, and plasmid copy numbers were 6.07×109 , 3.21×109 , 2.32×1010 , 8.11×108 , respectively. The plasmid yields were 55 ng.µL-1, 69 ng.µL-1, 164 ng.µL-1 and 41 ng.µL-1, respectively. Conclusion: Protein and plasmid yields of E. coli are variable in different concentrations of ampicillin and need to be optimized in newly designed expression systems. Protein and plasmid yield in the optimized concentration (200 µg.mL-1) was significantly (p < 0.01) higher than other doses.

Effect of Probiotic Soy Milk on Serum Levels of Adiponectin, Inflammatory Mediators, Lipid Profile, and Fasting Blood Glucose Among Patients with Type II Diabetes Mellitus.

Probiotic therapies are going to be an effective alternative therapeutic strategy in the treatment and management of diabetes. The mechanism behind the essential effects of probiotic therapies in diabetic patients was not fully understood. The objective of this study was to evaluate the effects of probiotic soy milk containing Lactobacillus planetarum A7 on inflammation, lipid profile, fasting blood glucose, and serum adiponectin among patients with type 2 diabetes mellitus. Forty patients with type 2 diabetes, at the age of 35-68 years old, were assigned to two groups in this randomized, double-blind, controlled clinical trial. The patients in the intervention group consumed 200 ml/day of probiotic soy milk containing L. planetarum A7 and those in control group consumed 200 ml/day of pure soy milk for 8 weeks. Serum TNF-α, C reactive protein, adiponectin, lipid profile, and fasting blood glucose were determined before and after intervention. In intervention group, serum adiponectin in pre- and post-treatment did not show any significant changes (2.52 ± 0.74 vs 2.84 ± 0.61, P = 0.658), as well as changes in serum TNF-α and C reactive protein (172.44 ± 5.7 vs 172.83 ± 7.6, P = 0.278, 4.2 ± 1.4 vs 4.5 ± 1.9, P = 0.765, respectively). Low-density cholesterol and high-density cholesterol changed significantly (P = 0.023, P = 0.017, respectively), but fasting blood glucose did not show any significant changes. The results of this study showed that consumption of probiotic soy milk and soy milk has no effect on serum adiponectin and inflammation, but it can change lipid profile among type 2 diabetic patients.

Expression of biologically active murine interleukin-18 in Lactococcus lactis.

The food-grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L. lactis to secrete murine IL-18. The mature murine IL-18 gene was inserted downstream of a nisin promoter in pNZ8149 plasmid and the construct was used to transform L. lactis NZ3900. The transformants were selected on Elliker agar and confirmed by restriction enzyme digestion and sequencing. The expression and secretion of IL-18 protein was verified by SDS-PAGE, western blotting and ELISA. The biological activity of recombinant IL-18 was determined by its ability to induce interferon (IFN)-γ production in L. lactis co-cultured with murine splenic T cells. The amounts of IL-18 in bacterial lysates and supernatants were 3-4 µg mL-1 and 0.6-0.7 ng mL-1, respectively. The successfully generated L. lactis strain that expressed biologically active murine IL-18 can be used to evaluate the possible therapeutic effects of IL-18 on mucosal surfaces.

Transforming growth factor beta-1 (TGF-ß1) gene single nucleotide polymorphisms (SNPs) and susceptibility to pre-eclampsia in Iranian women: A case-control study.

OBJECTIVES: Pre-eclampsia (PE) is a disorder of pregnancy characterized by high blood pressure and proteinuria. Transforming growth factor beta-1 (TGF-ß1) is an important replicated PE candidate gene, and few studies have evaluated the direct association of TGF-ß polymorphisms and risk to PE. The aim of this study was to investigate the association between three SNPs of TGF-ß1 and serum level of this cytokine in PE patients and controls. DESIGN AND METHODS: In this study the polymorphisms of the TGF-ß1 gene at the coding region, and positions 29T→C (Leu 10 Pro), 74G→C (Arg 25 Pro) and 788C→T (Thr 263 Ile) were studied in 123 PE and 120 normal subjects using PCR-restriction fragment length polymorphism PCR-(RFLP) and amplification refractory mutation system (ARMS)-PCR methods. Moreover, serum TGF-ß1 was determined by enzyme-linked immunosorbent assay (ELISA) technique. RESULTS: At positions 74G→C and 29T→C the genotypes and allele frequencies showed no significant differences between PE patients and normal controls (P=0.3 and P=0.5 respectively). While in the case of position 788C→T both genotypes and allele frequencies were significantly different between PE patients and controls (P=0.02). Haplotype analysis on three polymorphic sites showed no significant differences between PE and control individuals (P=0.8). TGC and CGC haplotypes were the most frequent in both studied groups. The mean serum TGF-ß1 level was significantly higher (62.73ng/ml) in PE patients compared with pregnant (47.01ng/ml) and non-pregnant (40.68ng/ml) control groups (P=0.0001). CONCLUSIONS: The results of this study suggest that TGF-ß1 gene 788C→T polymorphism is an important factor mediating the casual pathway of preeclampsia.

Augmented plasma adiponectin after prolonged fasting during ramadan in men.

BACKGROUND: Intermittent fasting during Ramadan entails major changes in metabolism and energy expenditure. This study sought to determine effect of the Ramadan fasting on serum levels of adiponectin and tumor necrosis factor-α (TNF-α) as two inter-related peptides involved in cells sensitivity to insulin and glucose metabolism. METHODS: Total of seventy healthy men, with age range equal or greater than 30, with at least three type2 diabetes mellitus (DM) risk factors were selected. Serum lipid profile, anthropometric indices and plasma glucose levels were determined using conventional methods. Also, serum adiponectin and TNF- α concentra-tions were assessed using Enzyme-linked Immunosorbent Assay. Data were analyzed by paired t-test. RESULTS: Ramadan fasting resulted in a significant increase of serum adiponectin (P< 0.000), fasting glucose (P< 0.000) and triglycride (P< 0.001). Body mass index was lowered during the fasting (P< 0.000). Finally, no remarkable decrease was found in serum TNF-α levels (P= 0.100). CONCLUSION: Ramadan fasting resulted in augmented adipo-nectin levels which may help in improving metabolic stress induced by insulin resistance in men with predisposing factors of type2 DM.

Promoter region polymorphisms in the transforming growth factor beta-1 (TGFß1) gene and serum TGFß1 concentration in preeclamptic and control Iranian women.

Preeclampsia (PE) is a pregnancy associated disorder characterized by hypertension and proteinuria, which causes neonatal and maternal morbidity and mortality. The Th1/Th2 cytokine paradigm of the immune adaptation in pregnancy is now expanded to include Th1/Th2/Th17 and regulatory T (Treg) cells. Among cytokines, TGFß1 has properties that justify evaluation of its role in PE etiopathology. In this investigation the polymorphisms of the TGFß1 gene at promoter region, positions -800G→A and -509C→T, were studied in 142 PE and 140 normal pregnant female subjects using PCR-RFLP. Additionally, serum TGFß1 was determined by ELISA. At position -800G→A genotypes and allele frequencies showed no significant differences between PE patients (GG 73.9%; GA 21.1%; AA 4.93%) and normal control (GG 70%; GA 28.6%; AA 1.4%) women. However the AA genotype at this position was more frequent in PE patients than in the control group. At -509C→T position, genotypes and allele frequencies showed no significant differences between PE patients and control individuals. The CC genotype at -509C→T position was more prevalent in PE patients than in the control group. The mean serum TGFß1 level was significantly higher (62.14 ng/ml) in PE patients compared with pregnant and non-pregnant control groups (and 47.01 and 40.68 ng/ml, respectively). In conclusion, the promoter region polymorphisms of TGFß1 may not be associated with PE, but serum levels of this cytokine may contribute to the etiopathology of PE.