Targeting Iron - Respiratory Reciprocity Promotes Bacterial Death.

Discovering new bacterial signaling pathways offers unique antibiotic strategies. Here, through an unbiased resistance screen of 3,884 gene knockout strains, we uncovered a previously unknown non-lytic bactericidal mechanism that sequentially couples three transporters and downstream transcription to lethally suppress respiration of the highly virulent P. aeruginosa strain PA14 - one of three species on the WHO's 'Priority 1: Critical' list. By targeting outer membrane YaiW, cationic lacritin peptide 'N-104' translocates into the periplasm where it ligates outer loops 4 and 2 of the inner membrane transporters FeoB and PotH, respectively, to suppress both ferrous iron and polyamine uptake. This broadly shuts down transcription of many biofilm-associated genes, including ferrous iron-dependent TauD and ExbB1. The mechanism is innate to the surface of the eye and is enhanced by synergistic coupling with thrombin peptide GKY20. This is the first example of an inhibitor of multiple bacterial transporters.

Novel mutation in efflux pump Rv1258c (Tap) gene in drug resistant clinical isolates of Mycobacterium tuberculosis in Iran.

INTRODUCTION: Tuberculosis (TB) remains a serious public health problem worldwide. Drug-resistant TB is considered a major and growing global threat. Despite the great variety of described mutations in Mycobacterium tuberculosis (MTB) resistance genes, the mechanisms of drug resistance are still controversial. Recently, a report on the role of efflux pump genes in drug resistance added to this complexity. Therefore, a thorough understanding of efflux pump genes in drug-resistant TB clinical isolates is needed. METHODOLOGY: We performed molecular analysis of the efflux pump gene (Rv1258c) in 33 drug-resistant and 20 drug-sensitive clinical MTB isolates by sequencing the amplicons' targets in both the forward and reverse directions. RESULTS: A novel mutation of the Rv1258c gene was identified at G442A (Ala148Thr) in rifampicin mono-resistant clinical strain, as compared to the H37Rv reference strain. In addition, a cytosine nucleotide insertion was found between the positions 580 and 581 (denominated Tap580) in two drug-sensitive strains at identical gene positions. CONCLUSIONS: These results indicated the possibility of mutation in the efflux pump genes and the important role of Tap efflux pump genes in drug-resistant MTB isolates. However, further research is required to determine the direct association of these mutations with resistant MTB.

The effect of increasing duration of family members' presence on sleep status in patients with acute coronary syndrome in cardiac care unit: A randomized controlled trial.

AIM: To investigate the effect of increasing duration of family members' presence on sleep status in patients with acute coronary syndrome (ACS) admitted to the cardiac care unit. DESIGN: Randomized controlled trial. METHODS: Ninety patients with ACS randomly assigned into two groups. No intervention was performed in control group. In the intervention group, the time of family members presence was changed from 1 h per day to 2 h per day from the second to the fourth day. Then, ST Mary's Hospital Sleep Questionnaire was completed by the patients every day during the study. RESULTS: The patients in the intervention group had statistically significantly better sleep status during the course of intervention compared to the control group. PATIENT OR PUBLIC CONTRIBUTION: The increasing duration of family members' presence can improve the sleep quality and quantity of ACS patients.

Crosstalk between tau protein autoproteolysis and amyloid fibril formation.

Tau cleavage has been shown to have a significant effect on protein aggregation. Tau truncation results in the formation of aggregation-prone fragments leading to toxic aggregates and also causes the formation of harmful fragments that do not aggregate. Thus, targeting proteolysis of tau would be beneficial for the development of therapeutics for Alzheimer's disease and related tauopathies. In this study, amino-terminal quantification and ThT fluorimetry were respectively used to analyze the kinetics of tau fragmentation and fibril formation. SDS-PAGE analysis of tau protein incubated with a disulfide-reducing agent demonstrated that the cysteines of tau have a crucial role in the fibrillation and autoproteolysis. However, the structures converted to amyloid fibrils were different with conformations that led to autoproteolysis. The quantification of the amino terminal indicated that the double-disulfide parallel structures formed in the presence of heparin did not have protease activity. The survey of possible tau disulfide-mediated dimer configurations suggested that the non-register single disulfide bound conformations were involved in the tau autoproteolysis process. Moreover, the inhibition of autoproteolysis resulted in the increment of aggregation rate; hence it seems that the tau auto-cleavage is the cellular defense mechanism against protein fibrillation.

Guidelines for an optimized differential centrifugation of cells.

Literature reviews reveal a significant deficiency in conceptual comprehension concerning centrifugation, a crucial step in both medical and research protocols. The arbitrary fluctuations in centrifugal forces present a potential threat to the reproducibility of results. To address this, we propose concise guidelines that integrate key factors such as temperature, osmolarity, fluid volume, and viscosity. These guidelines aim to enhance comprehension of optimal sedimentation conditions for cell suspensions. Additionally, we introduce a standardized protocol for determining the optimal RCF and centrifugation time. The goal is to maximize sedimentation efficiency while minimizing cell damage, contributing to a universally applicable and reproducible method in centrifugation practices.

Human exposure to pesticides and thyroid cancer: a worldwide systematic review of the literatures.

Thyroid cancer is considered as one of the most prevalent cancers in the world. Some pesticides can play a role as a potentially important risk factor in thyroid cancer by affecting thyroid morphology and thyroid hormone homeostasis. The aim of present study was to systematically review the available epidemiological evidence for human exposure to pesticides and thyroid cancer. Articles were searched in PubMed, Scopus and Web of Science by suitable keywords from January 2000 to May 2021. Standard techniques for systematic reviews were followed in the current study and results reported according to Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. Based on the inclusion and exclusion criteria, finally seven studies including four cohort studies and three case-control studies were reviewed. Organochlorines (OCPs) in more cases, Organophosphates (OPs) and Carbamates insecticides, herbicides and fungicides were the studied pesticides. Inconsistent results were reported in the surveyed articles on the OCPs. Two articles on the Carbamates (Carbaryl and Mancozeb) showed consistently an inverse association between exposure and thyroid cancer. Increased risk of thyroid cancer due to the exposure to the Malathion was reported in one article on the OPs. Due to the limited current knowledge about the effect of pesticides on thyroid cancer in humans, human health policies must be implemented to control individual's exposure to chemicals through using of botanical pesticides in agricultural. Also, more studies must be done to fill this gap of knowledge.

PVA-Based Nanofibers Containing Chitosan Modified with Graphene Oxide and Carbon Quantum Dot-Doped TiO2 Enhance Wound Healing in a Rat Model.

Electrospun nanofibrous constructs based on nanoparticles and biopolymers have recently been used in tissue engineering because of their similarity to the extracellular matrix in nature. In this study, electrospun chitosan-carbon quantum dot-titanium dioxide-graphene oxide (CS-CQD-TiO2-GO) nanofibrous mats were synthesized for use as wound dressings by the electrospinning method. To increase the biodegradation rate and water resistance, the fabricated nanofibrous mats were cross-linked. SEM images showed a uniform and coherent structure of CS-CQD-TiO2-GO nanocomposites and CS-CQD-TiO2-GO electrospun nanofibers mats. FTIR analysis, XRD pattern, SEM mapping, and EDS spectrum demonstrate the accuracy of the synthesis as well as the elemental and chemical structure of the nanofibrous mat. The water contact angle indicated that the nanofibrous mat had a hydrophilic property, which is essential for controlling wound exudates. The tensile strength and elongation tests showed that the nanofibrous mat has suitable mechanical properties for wound dressing, including significant flexibility and strength. Interestingly, antimicrobial testing illustrated that the fabricated nanofibrous mat had antibacterial activity against Gram-negative and Gram-positive bacteria. Appropriate cell viability and cytocompatibility of treated mouse fibroblast NIH3T3 cells with the nanofibrous mat were determined using an MTT assay. The animal study results confirmed the proper potential of the nanofibrous mat in wound dressing applications.

Autophagy in the normal and diseased cornea.

The cornea and covering tear film are together the 'objective lens' of the eye through which 80% of light is refracted. Despite exposure to a physically harsh and at times infectious or toxic environment, transparency essential for sight is in most cases maintained. Such resiliency makes the avascular cornea a superb model for the exploration of autophagy in the regulation of homeostasis with relevancy to all organs. Nonetheless, missense mutations and inflammation respectively clog or apparently overwhelm autophagic flux to create dystrophies much like in neurodegenerative diseases or further exacerbate inflammation. Here there is opportunity to generate novel topical therapies towards the restoration of homeostasis with potential broad application.

Significance of the coexistence of non-codon 315 katG, inhA, and oxyR-ahpC intergenic gene mutations among isoniazid-resistant and multidrug-resistant isolates of Mycobacterium tuberculosis: a report of novel mutations.

Tuberculosis (TB) is a global threat due to the emergence and spread of drug-resistant Mycobacterium tuberculosis (MTB). Isoniazid (INH) is the main antibiotic used for prevention and treatment of TB. Evidence shows that accumulated mutations can produce INH resistant (INHR) strains, resulting in the progression of multidrug-resistant (MDR) TB. Since point mutations in katG gene, inhA gene, and oxyR-ahpC region correlated with the INH resistance, in this study, we aimed to identify mutations in these three genes in INHR and MDR clinical isolates of MTB by Sanger DNA sequencing analysis. Thirty-three out of 438 isolates were resistant, including 66.7% INHR and 30.3% MDR isolates. In the katG gene, 68.2% INHR isolates had non-synonymous point mutations, mainly R463L (63.6%), and non-synonymous point mutation KatG L587P was seen in one of the MDR isolate. A novel silent substitution L649L was identified in the inhA gene of the MDR isolates. The oxyR-ahpC intergenic region g-88a common mutations (63.6%) in INHR and two distinct novel mutations were found at positions -76 and -77 of the oxyR-ahpC intergenic region. The coexistence of katG non-codon 315 with oxyR-ahpC intergenic region mutations was highly frequent in INHR 59.1% and MDR isolates 70%. Since mutations of all three genes 95.5% lead to the detection of INHR, they might be useful for molecular detection. Our results indicated the continuous evolution and region-specific prevalence of INH resistance. Overall, identification of new mutations in INH resistance can improve the available strategies for diagnosis and control of TB.

Comparison of the Effect of Nifedipine Alone and the Combination of Nifedipine and Sildenafil in Delaying Preterm Labor: A Randomized Clinical Trial.

Objective: Recently, sildenafil as a drug effective in relaxing smooth muscles can be used as an adjunct to delay the onset of uterine contractions and therefore the occurrence of preterm labor. The aim of this study was to evaluate the effect of nifedipine combination with sildenafil on preterm delivery compared with nifedipine alone. Materials and methods: This randomized double-blinded clinical trial was performed on pregnant women with a gestational age of 26-34 weeks with singleton pregnancy and symptoms of preterm delivery. The mothers were randomly assigned into two groups receiving nifedipine plus sildenafil or those receiving nifedipine alone. The time of delivery, maternal and neonatal complications were compared between the two groups. Results: Mothers who received the combination therapy experienced significantly lower preterm delivery within 72 hours of intervention compared to nifedipine alone (4.5% versus 27.3%, p = 0.002). The rate of delivery during the first 7 days after discharge was 7.6% and 31.8% in nifedipine plus sildenafil and nifedipine alone, respectively (P = 0.001). The prevalence of neonatal respiratory distress syndrome (RDS) as well as mean birth weight was higher in the nifedipine group alone. Treatment protocol with nifedipine and sildenafil compared with nifedipine alone was associated with a significant increase in preterm delivery delay (beta =-5.819, p = 0.001). Conclusion: The use of sildenafil in addition to nifedipine causes more delay in delivery in cases of preterm labor, followed by lower risk for RDS, reduces neonatal intensive care unit (NICU) admission, and preserves neonatal birth weight.

Molecular detection of Extended-Spectrum ß-lactamases (ESBLs) and biofilm formation in uropathogen Klebsiella pneumoniae in Iran.

Background: Uropathogenic Klebsiella pneumoniae is one of the well-kown uropathogens that have the main rule in biofilm formation. Increased prevalence of ESBL enzyme is one of the therapeutic problems. However, the aims of this study were to characterize the ability of biofilm formation and ESBL-producing isolates produced by urinary tract infection's K. pneumoniae to identify the prevalence of this type of infection in the studied area. Methods: Between the 500 nonrepetitive clinical isolates, 128 isolates were detected as K. pneumoniae. Biofilm production of these isolates was showed by Merrit and Christensen method. The standard Kirby-Bauer disk diffusion method was used for antimicrobial susceptibility testing. The phenotype ESBL was confirmed by double disc synergy test (DDST). Genotypic identification of ESBLs did by molecular detection. The statistical analysis was done using software IBM SPSS Statistics (SPSS Inc) and chi-square and Fisher exact tests. Results: The result of microtiter plate was observed and it was found that 86 (67.2%) isolates had weak biofilm, 24 (18.8%) moderate biofilm, and 18 (14.1%) strong biofilm. Also, 57 (44.5%) out of 128 isolates were diagnosed as MDR. The highest frequency of resistance was identified for cefotaxime 60 (46.9%) and tetracycline 60 (46.9%), and the lowest rate was for amikacin 16 (12.5%). The results of DDST showed 55 of 128 (43%) produced ESBL enzymes. PCR detection in ESBL-producing isolates showed contained blaTEM 33 of 55(63.1%), and blaVEB 13 of 55 (23% ). Also, 1 of 55 (2%) had both blaTEM and blaVEB . Also, 5 of 13 (38.4%) isolates that had the blaVEB gene were also MDR and had weak biofilm (8/13; 61.5%), intermediate biofilm (3/13; 23%), and strong biofilm (2/13; 15.4%). Conclusion: To decrease treatment complications and mortality rate of drug-resistant bacterial infections, rapid detection of ß-lactamases genes and evaluation of these properties and infection management programs can help to prevent the transmission of drug resistant-strains.

A comprehensive review of long non-coding RNAs in the pathogenesis and development of non-alcoholic fatty liver disease.

One of the most prevalent diseases worldwide without a fully-known mechanism is non-alcoholic fatty liver disease (NAFLD). Recently, long non-coding RNAs (lncRNAs) have emerged as significant regulatory molecules. These RNAs have been claimed by bioinformatic research that is involved in biologic processes, including cell cycle, transcription factor regulation, fatty acids metabolism, and-so-forth. There is a body of evidence that lncRNAs have a pivotal role in triglyceride, cholesterol, and lipoprotein metabolism. Moreover, lncRNAs by up- or down-regulation of the downstream molecules in fatty acid metabolism may determine the fatty acid deposition in the liver. Therefore, lncRNAs have attracted considerable interest in NAFLD pathology and research. In this review, we provide all of the lncRNAs and their possible mechanisms which have been introduced up to now. It is hoped that this study would provide deep insight into the role of lncRNAs in NAFLD to recognize the better molecular targets for therapy.

Angiotensin-Converting Enzyme Inhibitor Captopril: Does it Improve Renal Function in Lipopolysaccharide-induced Inflammation Model in Rats.

Renin-angiotensin system as an important regulator of renal function has also a major role in inflammation. In the present study, the effects of captopril on renal dysfunction, renal cytokine levels, and renal tissue oxidative damage were investigated in lipopolysaccharide (LPS)-induced inflammation model in rats. Treatment of five groups of the rats was carried out as follows: (1) saline as a control, (2) LPS 1 mg/kg, and (3-5) 10, 50, or 100 mg/kg captopril 30 min, respectively, before LPS. The treatments were given for 12 days. Finally, the animals were deeply anesthetized, the blood samples were obtained, and the renal tissues were removed and kept for biochemical measurements. Administration of LPS increased serum blood urea nitrogen and creatinine (P < 0.001). Pretreatment with all doses of captopril decreased these parameters (P < 0.001). LPS also increased interleukin-6 (IL-6), malondialdehyde, and nitric oxide metabolites in the renal tissues (P<0.05 - P < 0.001), which was prevented by captopril (P < 0.05 - P < 0.001). The total thiol concentration and superoxide dismutase and catalase activities in the kidney of the LPS group were lower than the control (P < 0.001), while they were enhanced when the animals were cotreated by captopril (P <0.01 - P < 0.001). The results of the present study showed that captopril improved renal function and attenuated tissue oxidative stress in LPS-induced inflammation model in rats.

Determination of morphine and oxymorphone in exhaled breath condensate samples: Application of microwave enhanced three-component deep eutectic solvent-based air-assisted liquid-liquid microextraction and derivatization prior to gas chromatography-mass spectrometry.

In this work, a microwave-enhanced air-assisted liquid-liquid microextraction method combined with gas chromatography-mass spectrometry has been developed for morphine and oxymorphone assessment in EBC samples. For this purpose, choline chloride-menthol-phenylacetic acid deep eutectic solvent (as an extraction solvent), butyl chloroformate (as a derivatization agent), and picoline (as a catalyst) are used. After performing predetermined extraction cycles in the microextraction method, the obtained cloudy solution is exposed to microwave irradiations to enhance extraction and derivatization efficiencies. The method provided low limits of detection (morphine 2.1 and oxymorphone 1.5 ng mL-1) and quantification (morphine 7.2 and oxymorphone 5.2 ng mL-1) in the EBC samples. The method had proper repeatability, accuracy, and stability expressed as relative standard deviations less than 5.1, 9, and 9%, respectively. The developed method was successfully used to determine morphine and oxymorphone concentrations in the EBC samples of addict patients.

Immunotherapy with IgY Antibodies toward Outer Membrane Protein F Protects Burned Mice against Pseudomonas aeruginosa Infection.

Burn patients with multidrug-resistant Pseudomonas aeruginosa infections commonly suffer from high morbidity and mortality, which present a major challenge to healthcare systems throughout the world. Outer membrane protein F (OprF), as a main outer membrane porin, is required for full virulence expression of P. aeruginosa. The aim of this study was to evaluate the protective efficacy of egg yolk-specific antibody (IgY) raised against recombinant OprF (r-OprF) protein in a murine burn model of infection. The hens were immunized with r-OprF, and anti-r-OprF IgY was purified using salt precipitation. Groups of mice were injected with different regimens of anti-OprF IgY or control IgY (C-IgY). Infections were caused by subcutaneous injection of P. aeruginosa strain PAO1 at the burn site. Mice were monitored for mortality for 5 days. The functional activity of anti-OprF IgY was determined by in vitro invasion assays. Immunotherapy with anti-OprF IgY resulted in a significant improvement in the survival of mice infected by P. aeruginosa from 25% to 87.5% compared with the C-IgY and PBS. The anti-OprF IgY decreased the invasion of P. aeruginosa PAO1 into the A549. Passive immunization with anti-OprF IgY led to an efficacious protection against P. aeruginosa burn infection in the burn model.

The protective effect of Nigella sativa extract on lung inflammation and oxidative stress induced by lipopolysaccharide in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Oxidative stress during inflammation can increase inflammation and damage tissue. Nigella sativa L. (NS) showed many pharmacological properties including antioxidant and anti-inflammatory activities. AIM OF THE STUDY: In this study, the preventive effect of NS on lung inflammation and oxidative stress induced by lipopolysaccharide (LPS) in the rats was investigated. MATERIALS AND METHODS: Male rats were assigned to: Control, LPS (1 mg/kg, i.p.), LPS + NS (100, 200, 400 mg/kg, i.p.), (10 per group). Saline (1 ml/kg) was intra-peritoneal (i.p.) injected instead of LPS in the rats of the control group. LPS dissolved in saline and injected i.p. daily for 14 days. Treatment with NS extracts started two days before LPS administration and treatment continued during LPS administration. White blood cells (WBC), total and differential as well as oxidative stress index in bronchoalveolar fluid (BALF) and serum, TGF-ß1, IFN-γ, PGE2, and IL-4 levels in the BALF and lung histopathology were examined. RESULTS: LPS administration increased total WBC, eosinophils, neutrophils, basophils, and monocytes counts as well as oxidative stress markers in the BALF and serum as well as TGF-ß1, IFN-γ, PGE2, IL-4 levels in the BALF and pathological changes of the lung tissue. All of these effects were reduced by NS extract treatment dose-dependently. CONCLUSION: These results suggested the protective effects of NS extract on lung inflammation and oxidative stress as well as its effect on lung pathology induced by LPS dose-dependently.

The effects of captopril on lipopolysaccharide-induced sickness behaviors in rats.

Neuro-immune mediators play an important role in the development of sickness behaviors. In the present study, the effect of captopril on sickness behaviors caused by lipopolysaccharide (LPS) was studied in the rats. The animals were randomized into the following groups: control, sham, 10 mg kg-1 captopril - LPS (Capto 10-LPS), 50 mg kg-1 captopril - LPS (Capto 50-LPS), and 100 mg kg-1 captopril - LPS (Capto 100-LPS). Behavioral tests including open-field (OF), elevated plus maze (EPM) and forced swimming (FS) test were performed, and the serum level of interleukin-6 (IL-6) was assessed. In OF, the number of crossings in the central zone in Capto 10-LPS, Capto 50-LPS, and Capto 100-LPS groups was higher than that of the sham group. In EPM, the open arm entry numbers in the sham group were lower compared to the control group. Furthermore, pretreatment by captopril increased the entries to the open arms. In FS test, the immobility time of the sham group was longer than that of the control group. In Capto 10-LPS, Capto 50-LPS, and Capto 100-LPS groups, immobility was shorter compared to the sham group. In addition, the IL-6 level was higher in the sham group compared to the control group, and treatment with 50 and 100 mg kg-1 of captopril restored the IL-6 level in comparison with the sham group. Results confirmed that pretreatment with captopril ameliorated LPS-caused sickness behaviors and attenuated IL-6 as an inflammatory marker in the rats.

Anti-PcrV IgY antibodies protect against Pseudomonas aeruginosa infection in both acute pneumonia and burn wound models.

Pseudomonas aeruginosa is a common nosocomial pathogen in burn patients, and rapidly acquires antibiotic resistance; thus, developing an effective therapeutic approach is the most promising strategy for combating infection. Type III secretion system (T3SS) translocates bacterial toxins into the cytosol of the targeted eukaryotic cells, which plays important roles in the virulence of P. aeruginosa infections in both acute pneumonia and burn wound models. The PcrV protein, a T3SS translocating protein, is required for T3SS function and is a well-validated target in animal models of immunoprophylactic strategies targeting P. aeruginosa. In the present study, we evaluated the protective efficacy of chicken egg yolk antibodies (IgY) raised against recombinant PcrV (r-PcrV) in both acute pneumonia and burn wound models. R-PcrV protein was generated by expressing the pcrV gene (cloned in pET-28a vector) in E. coli BL-21. Anti-PcrV IgY was obtained by immunization of hen. Anti-PcrV IgY induced greater protection in P. aeruginosamurine acute pneumonia and burn wound models than control IgY (C-IgY) and PBS groups. Anti-PcrV IgY improved opsonophagocytic killing and inhibition of bacterial invasion of host cells. Taken together, our data provide evidence that anti-PcrV IgY can be a promising therapeutic candidate for combating P. aeruginosa infections.

Memory enhancing effect of Nigella Sativa hydro-alcoholic extract on lipopolysaccharide-induced memory impairment in rats.

In this study, the effects of Nigella Sativa (NS) hydro-alcoholic extract on lipopolysaccharide (LPS)-induced learning and memory impairments, hippocampal cytokine levels, and brain tissues oxidative damage were investigated in rats. The rats were grouped and treated: (1) control (saline), (2) LPS (1 mg/kg i.p.), and (3-5) 100, 200, or 400 mg/kg NS hydro-alcoholic extract 30 min before LPS injection. The treatment was started since 6 days before the behavioral experiments and continued during the behavioral tests (LPS injection 2 h before each behavioral experiment). Finally, the brains were removed for biochemical assessments. In Morris water maze (MWM) test, LPS increased the escape latency and traveled path compared to control group, whereas all doses of NS hydro-alcoholic extract decreased them compared to LPS group. In passive avoidance (PA) test, the latency to enter the dark compartment in LPS group was shorter than control group while in all treated groups it was longer than LPS group. LPS increased tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malondialdehyde (MDA), and nitric oxide (NO) metabolites, and decreased thiol content, superoxide dismutase (SOD), and catalase (CAT) in the hippocampal tissues compared to control group while NS hydro-alcoholic extract decreased MDA and NO metabolites and increased thiol content, SOD, and CAT compared to LPS group. Findings of the current study indicated that the hydro-alcoholic extract of NS improved the LPS-induced learning and memory impairments induced by LPS in rats by improving hippocampal cytokine levels and brain tissues oxidative damage.