An update on alternative therapy for Escherichia coli causing urinary tract infections; a narrative review.

BACKGROUND: Urinary tract infections (UTIs) are the most common type of nosocomial infection and severe health issues because of the difficulties and frequent recurrence. Today, alternative methods such as sonodynamic therapy (SDT), photodynamic therapy (PDT) and herbal materials use for treating infections like UTI in many countries. METHOD: We conducted searches of the biomedical databases (Google Scholar, Scopus, PubMed, and Web of sciences) to identify related studies from 2008 to 2023. RESULT: SDT aims to use ultrasound to activate a sonosensitizer, which causes a biological effect by raising reactive oxygen species (ROS). When bacteria are exposed to ROS, several important effects occur: oxidative damage, DNA damage, protein dysfunction etc. SDT with herbal medicine significantly reduced the number of colony-forming units and bactericidal activity for Klebsiella pneumonia and E. coli. PDT is a promising treatment for cancer and microbial infections, combining a photosensitiser, light and tissue molecular oxygen. It involves a photosensitizer, light source, and oxygen, with variations affecting microbial binding and bactericidal activity. Factors affecting antibacterial properties include plant type, growing conditions, harvesting, and processing. This review highlights the recent advancements in sonodynamic, photodynamic, herbal, and bio-material-based approaches in the treatment of E. coli infections. CONCLUSIONS: These alternative therapies offer exciting prospects for addressing UTIs, especially in cases where traditional antibiotic treatments may be less effective. Further research and clinical studies are warranted to fully explore the potential of these innovative treatment modalities in combating UTIs and improving patient outcomes.

Molecular characterization of ESBL-producing uropathogenic Escherichia coli isolates among kidney transplant patients: Emergence and spread of B2-ST131 clone type.

This study was conducted to identify the distribution of virulence determinants in uropathogenic Escherichia coli (UPEC) isolates obtained from kidney transplant (KTP) and non-transplant patients (non-KTP) with urinary tract infections (UTI). Additionally, the (GTG)5 fingerprinting technique was used to investigate the genetic diversity of Extended-Spectrum B-Lactamase (ESBL)-positive isolates. In this case-control study, 111 urine isolates were obtained from non-KTPs and KTPs, respectively. The presence of genetic markers encoding adhesion proteins, toxins and major E. coli phylogroups was assessed through PCR amplification. Molecular typing of ESBL-positive UPEC strains was performed using (GTG)5 fingerprinting and Multilocus sequence typing (MLST) techniques. Overall, 65 and 46 UPEC isolates were obtained from non-KTPs and KTPs, respectively. Among the studied isolates, traT (85.6%) gene was the most frequently observed virulence gene, followed by kpsMT (49.5%). Using the 80% cut-off point, all the 35 UPEC isolates were classified into four major clusters, namely A, B, C, and D. The majority of the Sequence Type (ST) 131 isolates belonged to cluster A. Additionally, three ST1193 isolates belonged to cluster A and phylogroup B2. Moreover, ST38, ST131 and ST10 were in different cluster. In general, we observed significant differences in the papA, ompT, sat, and vat genes between KTPs and non-KTPs. Furthermore, since all the isolates carried one or more virulence factors (VFs), these findings are concerning in the context of managing UTIs caused by the UPEC strain. Additionally, the distribution of ST and Clonal Complex (CC) among isolates in the main clusters revealed significant differences between MLST and (GTG)5 fingerprinting analysis.

Modulatory effects of quercetin on histological changes, biochemical and oxidative stress of rat placenta induced by inhalation exposure to crude oil vapor.

The inhalation exposure to crude oil vapor (COV) has been shown to have adverse effects on the placenta and fetal development. The modulatory effects of quercetin (QUE) as a natural phenolic compound with antioxidant properties are promising for the protection of placental structure. This study aimed to investigate the modulatory role of QUE in mitigating histopathological damage, oxidative stress, and biochemical alteration in the placenta of COV-exposed pregnant rats. Forty-eight pregnant rats were divided into eight groups (days 15 and 20) as follows: 1-2) Control groups, 3-4) COV groups, 5-6) COV+QUE groups, and 7-8) QUE-treated groups (50 mg/kg). The inhalation method was used to expose pregnant rats to COV, and QUE was administered orally. On the 15th and 20th days of gestation, placental tissue was analyzed using PAS and H&E staining and immunohistochemistry. The expression of the caspase-3 gene and oxidative stress biomarkers including TAC, CAT, MDA, GPx, and SOD were investigated in the placental tissue. The COV significantly decreased the weight, diameter, and thickness of the placenta as well as the thickness of the junctional zone and labyrinth and the number of trophoblast giant cells in 15- and 20-day-old placentas (P<0.05). Also, COV significantly increased placental expression of caspase-3 and the oxidative stress biomarkers (P<0.05). The administration of QUE along with exposure to COV reduced morphometric and histological alteration, oxidative stress, and caspase-3 expression (P<0.05). Our findings indicated that QUE in COV-exposed pregnant rats can prevent placental histopathological alternations by increasing the activity of the antioxidant system.

CRISPR Typing of Clinical Strains of Salmonella spp. Isolated in Tehran, Iran.

Background: Salmonella is one of the most leading causes of food-born infection and death among infants and people with the poor immunity system. Because Salmonella spp. have diversity in the genome composition and pathogenicity, access to rapid identification and genotyping is necessary to control of salmonellosis. The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) typing is a genotyping method that checks these variable sequences in the bacterial genome in a specific species. This study aimed to differentiate Salmonella strains using CRISPR region. Methods: Salmonella isolates, previously identified via standard microbiological and molecular tests, were subjected to the study. Bacterial DNA was extracted and PCR was done using specific primers. The different PCR products were sequenced and the repeats patterns were used to identify additional or degenerate repeat clusters in the CRISPR region. All different sequences were analyzed using CRISPRtionary tool for dendrogram generation using the binary file. Results: Overall, 119 strains of various Salmonella serovars were used. The result showed unique CRISPR and diversity in spacer both in sequence and the number. Analysis of the extracted sequence and band patterns illustrated that, except for S. infantis, both S. enteritidis and S. typhimurium isolates were classified as a separate cluster. Conclusion: CRISPR genotyping could provide serotype/spacers dictionary and it is performed at low cost and high speed in comparison to the other typing methods. Therefore, the assessment of CRISPR and spacer content can be considered as a powerful and practical discriminatory method for subtyping of Salmonella isolates.

Antibacterial and antibiofilm effects of green synthesized selenium nanoparticles on clinical Klebsiella pneumoniae isolates.

Nanotechnology covers many disciplines, including the biological sciences. In this study, selenium nanoparticles (Se-NPs) were synthesized using Artemisia annua extract and investigated against clinical strains of klebsiella pneumoniae (K. pneumoniae) for their anti-biofilm effects. In this experimental study, from May 1998 to September 1998, 50 clinical samples of blood, urine, and sputum were collected, and K. pneumoniae strains were isolated using microbiological methods. Subsequently, the antibacterial effects of Se-NPs at concentrations of 12-25-50-100/5-6/3-25/125 µg/mL were studied. Finally, biofilm-producing strains were isolated, and the expression of mrkA biofilm gene was studied in real-time strains treated with Se-NPs using real-time polymerase chain reaction (PCR). Out of 50 clinical samples, 20 strains of K. pneumoniae were isolated. Minimum inhibitory concentration (MIC) results of Se-NPs showed that Se-NPs were capable of significant cell killing. Real-time PCR results also showed that mrkA gene expression was significantly reduced in strains treated with Se-NPs. According to this study, Se-NPs could reduce bacterial growth and biofilm formation, therefore, could be considered a candidate drug in the medical application for infections caused by K. pneumoniae.

Protective effect of quercetin on fetal development and congenital skeletal anomalies against exposure of pregnant Wistar rats to crude oil vapor.

BACKGROUND: Epidemiological evidence indicates a relationship between maternal exposure to crude oil vapors (COV) during pregnancy and adverse pregnancy outcomes. Quercetin (QUE) is a plant flavonoid with purported antioxidant and anti-inflammatory effects, which has been shown to prevent birth defects. This study was aimed to investigate the protective role of QUE on fetal development and congenital skeletal anomalies caused by exposure of pregnant rats to COV. METHODS: Twenty-four pregnant Wistar rats were randomly categorized into four groups of control, COV, COV + QUE, and QUE (50 mg/kg). The inhalation method was used to expose pregnant rats to COV from day 0 to 20 of pregnancy, and QUE was administered orally during this period. On day 20 of gestation, the animals were anesthetized and a laparotomy was performed, and then the weight and crown rump length (CRL) of the fetuses were determined. Skeletal stereomicroscopic evaluations of fetuses were performed using Alcian blue/Alizarin red staining method, and the expression of osteogenesis-related genes (Runx2 and BMP-4) was evaluated using qPCR. RESULTS: This study showed that prenatal exposure to COV significantly reduced fetal weight and CRL, and expression of Runx2 and BMP-4 genes. Moreover, COV significantly increased the incidence of congenital skeletal anomalies such as cleft palate, spina bifida and non-ossification of the fetal bones. However, administration of QUE with exposure to COV improved fetal bone development and reduced congenital skeletal anomalies. CONCLUSION: QUE can ameliorate the teratogenic effects of prenatal exposure to COV by increasing the expression of osteogenesis-related genes.

Biochemical, Serological and Immunological Tests for Diagnosis of Coronavirus Disease 2019 (COVID-19) Patients: A Comprehensive Systematic Review and Meta-analysis.

BACKGROUND: Recent studies have suggested the role of primary laboratory tests in addition to clinical symptoms for patients suspected to have coronavirus disease 2019 (COVID-19), which play a significant role in the diagnosis of COVID-19. However, the results of these studies are contradictory. The present study was conducted to evaluate biochemical, serological, and immunological tests for the diagnosis of COVID-19 patients. METHODS: This study was presented in accordance with the PRISMA protocol. This protocol is registered with the code CRD42019145410 in PROSPERO. We conducted a comprehensive literature search in databases, including Web of Science, PubMed/Medline, CINAHL Scopus, Cochrane Library, EMBASE, Science Direct, and EBSCO to find citations from the beginning of January 2019 until the beginning of April 2020 without any restrictions. RESULTS: Finally, 51 studies, including 5,490 COVID-19 patients, were included in the present metaanalysis. The prevalence of different factors observed in laboratory findings was as follows: the prevalence of lymphopenia in patients with COVID-19 accounted for 51.6% (95% CI: 44.0-59.1), elevated C-reactive protein (CRP) was 63.6% (95% CI: 57.0-69.8), elevated erythrocyte sedimentation rate (ESR) was 62.5% (95% CI: 50.1-73.5), elevated tumor necrosis factor alpha (TNFα) was 28.7% (95% CI: 9.0-62.1), elevated serum amyloid-A level was 74.7% (95% CI: 50.0-89.7), elevated procalcitonin level was 72.6% (95% CI: 58.1-83.5), elevated interleukin-6 level was 59.9% (95% CI: 48.2-70.5), reduced CD3 level was 68.3% (95% CI: 50.1-82.2), reduced CD4 level was 62.0% (95% CI: 51.1- 71.6), elevated lactate dehydrogenase (LDH) level accounted for 53.1% (95% CI: 43.6-62.4), elevated brain natriuretic peptide (BNP) accounted for 48.9% (95% CI: 30.4-67.7), reduced albumin and reduced pre-albumin levels in patients with COVID-19 were estimated to be 54.7% (95% CI: 38.1-70.2) and 49.0% (95% CI: 26.6-71.8), and D-dimer level was 44.9% (95% CI: 31.0-59.6). CONCLUSION: The results show lymphopenia, elevated ESR level, elevated CRP level, elevated serum amyloid-A, elevated TNFα, elevated procalcitonin level, elevated interleukin-6 level, reduced CD3, reduced CD4, elevated BNP, elevated LDH, reduced albumin, reduced pre-albumin, and elevated Ddimer levels as the most common findings at the time of admission.

Antiproliferative effects of mesenchymal stem cells carrying Newcastle disease virus and Lactobacillus Casei extract on CT26 Cell line: synergistic effects in cancer therapy.

BACKGROUND AND AIMS: Colorectal Cancer (CRC) is a frequent malignancy with a high mortality rate. Specific inherited and environmental influences can affect CRC. Oncolytic viruses and bacteria in treating CRC are one of the innovative therapeutic options. This study aims to determine whether mesenchymal stem cells (MSCs) infected with the Newcastle Disease Virus (NDV) in combination with Lactobacillus casei extract (L. casei) have a synergistic effects on CRC cell line growth. MATERIALS AND METHODS: MSCs taken from the bone marrow of BALB/c mice and were infected with the 20 MOI of NDV. Then, using the CT26 cell line in various groups as a single and combined treatment, the anticancer potential of MSCs containing the NDV and L. casei extract was examined. The evaluations considered the CT26 survival and the rate at which LDH, ROS, and levels of caspases eight and nine were produced following various treatments. RESULTS: NDV, MSCs-NDV, and L. casei in alone or combined treatment significantly increased apoptosis percent, LDH, and ROS production compared with the control group (P˂0.05). Also, NDV, in free or capsulated in MSCs, had anticancer effects, but in capsulated form, it had a delay compared with free NDV. The findings proved that L. casei primarily stimulates the extrinsic pathway, while NDV therapy promotes apoptosis through the activation of both intrinsic and extrinsic apoptosis pathways. CONCLUSIONS: The results suggest that MSCs carrying oncolytic NDV in combination with L. casei extract as a potentially effective strategy for cancer immunotherapy by promoting the generation of LDH, ROS, and apoptosis in the microenvironment of the CT26 cell line.

Antibiofilm effect of melittin alone and in combination with conventional antibiotics toward strong biofilm of MDR-MRSA and -Pseudomonas aeruginosa.

Introduction: Multidrug-resistant (MDR) pathogens are being recognized as a critical threat to human health if they can form biofilm and, in this sense, biofilm-forming MDR-methicillin resistant Staphylococcus aureus (MRSA) and -Pseudomonas aeruginosa strains are a worse concern. Hence, a growing body of documents has introduced antimicrobial peptides (AMPs) as a substitute candidate for conventional antimicrobial agents against drug-resistant and biofilm-associated infections. We evaluated melittin's antibacterial and antibiofilm activity alone and/or in combination with gentamicin, ciprofloxacin, rifampin, and vancomycin on biofilm-forming MDR-P. aeruginosa and MDR-MRSA strains. Methods: Antibacterial tests [antibiogram, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC)], anti-biofilm tests [minimum biofilm inhibition concentration (MBIC), and minimum biofilm eradication concentration (MBEC)], as well as synergistic antibiofilm activity of melittin and antibiotics, were performed. Besides, the influence of melittin alone on the biofilm encoding genes and the cytotoxicity and hemolytic effects of melittin were examined. Results: MIC, MBC, MBIC, and MBEC indices for melittin were in the range of 0.625-5, 1.25-10, 2.5-20, and 10-40 µg/ml, respectively. The findings found that the combination of melittin AMP with antibiotics was synergistic and fractional biofilm inhibitory concentration index (FBICi) for most tested concentrations was <0.5, resulting in a significant reduction in melittin, gentamicin, ciprofloxacin, vancomycin, and rifampin concentrations by 2-256.4, 2-128, 2-16, 4-64 and 4-8 folds, respectively. This phenomenon reduced the toxicity of melittin, whereby its synergist concentration required for biofilm inhibition did not show cytotoxicity and hemolytic activity. Our findings found that melittin decreased the expression of icaA in S. aureus and LasR in P. aeruginosa genes from 0.1 to 4.11 fold for icaA, and 0.11 to 3.7 fold for LasR, respectively. Conclusion: Overall, the results obtained from our study show that melittin alone is effective against the strong biofilm of MDR pathogens and also offers sound synergistic effects with antibiotics without toxicity. Hence, combining melittin and antibiotics can be a potential candidate for further evaluation of in vivo infections by MDR pathogens.

Investigate the immunogenic and protective effect of trivalent chimeric protein containing IpaD-StxB-TolC antigens as a vaccine candidate against S. dysenteri and S. flexneri.

BACKGROUNDS: Shigella spp. causes bloody diarrhea and leads to death, especially in children. Chimeric proteins containing virulence factors can prevent Shigella infection. The purpose of this study is to investigate the immunogenic and protective effect of trivalent chimeric protein containing IpaD-StxB-TolC antigens against shiga toxin, S. dysenteri and S. flexneri in vitro and in vivo conditions. METHODS: Recombinant vector was transferred to E. coli BL21. The expression of the chimeric protein was confirmed by SDS PAGE and purified using the Ni-NTA column. Mice were immunized with recombinant protein and antibody titer was evaluated by ELISA. 10, 25 and 50 LD50 of Shiga toxin neutralization was evaluated in vitro (Vero cell line) and in vivo conditions. Also, the challenge of immunized mice with 10, 25 and 50 LD50 of S. dysentery and S. flexneri was done. RESULTS: The expression and purification of the recombinant protein with 60.6 kDa was done. ELISA showed increased antibody titer against the chimeric protein. MTT assay indicated that 1/8000 dilution of the sera had a 51% of cell viability against the toxin in Vero cell line. The challenge of mice immunized with toxin showed that the mice had complete protection against 10 and 25 LD50 of toxin and had 40% survival against 50 LD50. Mice receiving 10 and 25 LD50 of S. dysenteri and S. flexneri had 100% protection and in 50 LD50 the survival rate was 60 and 50%, respectively. Organ burden showed that the amount of bacterial colonization in immunized mice was 1 × 104 CFU/mL, which was significantly different from the control group. CONCLUSION: This study showed that chimeric proteins can create favorable immunogenicity in the host as vaccine candidates.

The Effect of Cytotoxicity and Antimicrobial of Synthesized CuO NPs from Propolis on HEK-293 Cells and Lactobacillus acidophilus.

Background: Drug resistance is currently possible anywhere in the world. Due to the discovery of antimicrobials, medicine, and health have made tremendous advances over the past several decades. Aim: This research evaluated the antimicrobial and cytotoxicity effects of green synthesis of copper oxide nanoparticles (CuO NPs) on Lactobacillus acidophilus and human embryonic kidney 293 cells (HEK). Method and Materials. Propolis was sampled and extracted. Green synthesis of CuO NPs was synthesized and characterized using SEM, TEM, DLS, BET, and zeta potential methods. L. acidophilus (ATCC 4356) was used, and the antimicrobial tests were carried out at different concentrations (10≥ mg/ml). Moreover, the cytotoxicity was evaluated using an MTT assay on human embryonic kidney 293 cells (HEK). Results: Synthesized CuO NPs using propolis extracts from Khalkhal (sample 1) and Gillan (sample 2) showed -13.2 and -14.4 mV, respectively. The hydrodynamic sizes of well-dispersed samples 1 and 2 were 3124.9 nm and 1726.7 nm, respectively. According to BET analysis, samples 1 and 2 had 5.37 and 8.45 m2/g surface area, respectively. The surface area was decreased due to the addition of propolis extract, and the pore size was increased. CuO NPs of samples 1 and 2 were visible on SEM images with diameters ranging from 75 to 145 nm and 120 to 155 nm, respectively. Based on TEM analysis, the size of CuO particles was increased in samples 1 and 2. CuO NPs particles had narrow size distributions with evenly dispersed NPs on all sides. The cell viability of the CuO NPs of samples 1 and 2 after 24, 48, and 72 hours was greater than 50%. As a result of the MIC and MBC tests, it was determined that samples 1 and 2 had the same effect against L. acidophilus (0.0024 mg/ml). Biofilm formation and degradation of sample 1 were more efficient against L. acidophilus. Conclusion: There was no evidence of cytotoxicity in the samples. In addition, results showed that the green synthesized CuO NPs from Khalkhal propolis were effective against L. acidophilus. Thus, the green synthesized CuO NPs from Khalkhal propolis were the best candidates for clinical application.

Klebsiella pneumoniae: an update on antibiotic resistance mechanisms.

Klebsiella pneumoniae colonizes mucosal surfaces of healthy humans and is responsible for one third of all Gram-negative infections in hospitalized patients. K. pneumoniae is compatible with acquiring antibiotic resistance elements such as plasmids and transposons encoding various ß-lactamases and efflux pumps. Mutations in different proteins such as ß-lactamases, efflux proteins, outer membrane proteins, gene replication enzymes, protein synthesis complexes and transcription enzymes also generate resistance to antibiotics. Biofilm formation is another strategy that facilitates antibiotic resistance. Resistant strains can be treated by combination therapy using available antibiotics, though proper management of antibiotic consumption in hospitals is important to reduce the emergence and proliferation of resistance to current antibiotics.

The comparative effects of erythromycin and amikacin on acute respiratory Pseudomonas aeruginosa infection.

BACKGROUND: One of the most common causes of pneumonia is Pseudomonas aeruginosa (P. aeruginosa). As with other microbial pathogens, this bacterium tends to develop resistance to various antibiotics. Amikacin and erythromycin, which are from the aminoglycoside and macrolide antibiotic families, are used to treat respiratory infections caused by P. aeruginosa. OBJECTIVES: This study explored whether amikacin, erythromycin or a combination of both works better against P. aeruginosa acute lung infection. METHODS: For this study, 32 rats were used. The trachea of rats was exposed aseptically and their lung was infected with P. aeruginosa through trachea. Then, according to the group, they received amikacin, erythromycin or a combination of both for 1 week. Finally, they were euthanised on the 3rd and 7th days post-infection. The macroscopic and microscopic evaluations of the lungs, kidney and liver were performed. The right lung was collected for in vivo bacteriological analysis. RESULTS: The amikacin group (A group) had a statistically significantly lower macroscopic and microscopic scores than the other groups (p < 0.05). In vivo bacteriological test revealed that the A group had significantly lower lung bacterial load (p < 0.05). CONCLUSIONS: In summary, it was concluded that amikacin could help alleviate the respiratory infection caused by P. aeruginosa solely, and it was more effective than erythromycin.

Small-molecule metabolites in SARS-CoV-2 treatment: a comprehensive review.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has quickly spread all over the world. In this respect, traditional medicinal chemistry, repurposing, and computational approaches have been exploited to develop novel medicines for treating this condition. The effectiveness of chemicals and testing methods in the identification of new promising therapies, and the extent of preparedness for future pandemics, have been further highly advantaged by recent breakthroughs in introducing noble small compounds for clinical testing purposes. Currently, numerous studies are developing small-molecule (SM) therapeutic products for inhibiting SARS-CoV-2 infection and replication, as well as managing the disease-related outcomes. Transmembrane serine protease (TMPRSS2)-inhibiting medicinal products can thus prevent the entry of the SARS-CoV-2 into the cells, and constrain its spreading along with the morbidity and mortality due to the coronavirus disease 2019 (COVID-19), particularly when co-administered with inhibitors such as chloroquine (CQ) and dihydroorotate dehydrogenase (DHODH). The present review demonstrates that the clinical-stage therapeutic agents, targeting additional viral proteins, might improve the effectiveness of COVID-19 treatment if applied as an adjuvant therapy side-by-side with RNA-dependent RNA polymerase (RdRp) inhibitors.

Anatomical study of arterial arrangement of the spinal cord in Syrian hamsters (Mesocricetus auratus).

This study aimed to investigate the arterial arrangement of the spinal cord in Syrian hamsters, and to identify differences and similarities to humans and experimental animals that are mostly used as models in studies into ischemic spinal cord injuries. This observational anatomical study was conducted on 20 adult Syrian hamsters using dissection and corrosion casting technique. The general anatomy of the arterial blood supply of spinal cord was obtained and noted as follows: (1) high variability in the level of the origin of right and left vertebral arteries, (2) the independent origin of dorsal intercostal arteries, (3) origin of lumbar arteries as a common trunk with right and left divisions, (4) presence of dorsal and ventral radicular branches in both sides of the cervical spinal cord with almost the same frequency, (5) greater presence of ventral and dorsal radicular branches in the left side of the thoracolumbar spinal cord, (6) two dorsal spinal arteries originating from the posterior inferior cerebellar arteries and extending to the caudal end of the thoracic spinal cord, (7) continuous ventral spinal artery originating from both vertebral arteries and extending to the conus medularis, and (8) presence of the artery of Adamkiewicz in the thoracic region of the spinal cord. From comparative anatomy viewpoints, the arteries supplying the spinal cord of Syrian hamsters exhibit many similarities with humans, laboratory rodents, and rabbits in many aspects compared to dogs, cats, and pigs. Overall, Syrian hamsters can be used as a proposed model in experimental studies of the spinal cord ischemia.

Helicobacter pylori Infection: Conventional and Molecular Strategies for Bacterial Diagnosis and Antibiotic Resistance Testing.

Helicobacter pylori infection is a common health problem, which can cause gastric and extragastric diseases. Accurate detection of H. pylori is critical for appropriate patient management and bacterial eradication. In this regard, there are several methods for the diagnosis of H. pylori infection, which are classically divided into two major groups of invasive and non-invasive methods. Invasive methods, such as endoscopy, histology, and relative culture are less preferred due to their operational difficulties. By contrast, non-invasive methods, such as urea breath test, (UBT) are clinically preferred. Moreover, molecular methods, including polymerase chain reaction (PCR)-based methods, next-generation sequencing (NGS), and DNA microarray, have shown good sensitivity and specificity, and are considered helpful in H. pylori diagnosis. These methods have also increasingly concentrated on the detection of bacterial antibiotic resistance patterns. Besides, point of care (POC) devices play an important role in H. pylori diagnosis, mainly by shortening the time to the result and by making the test available at the bedside or at remote care centres. Biosensors, including aptasensors, have shown to be more appropriate tools because of their low limit of detection, high selectivity, fast response, and ease of handling. Finally, new emerging techniques, like MALDI-TOF MS, have been considered as fast methods with high degree of accuracy and sensitivity to identify and differentiate new species of H. pylori. The current article reviews the most recent developments in invasive, noninvasive, and molecular approaches for the diagnosis of H. pylori infections. Moreover, the application of emerging techniques, including MALDI-TOF MS and recently developed POCs, and biomarker-based methods, is discussed.

Synthetic materials in craniofacial regenerative medicine: A comprehensive overview.

The state-of-the-art approach to regenerating different tissues and organs is tissue engineering which includes the three parts of stem cells (SCs), scaffolds, and growth factors. Cellular behaviors such as propagation, differentiation, and assembling the extracellular matrix (ECM) are influenced by the cell's microenvironment. Imitating the cell's natural environment, such as scaffolds, is vital to create appropriate tissue. Craniofacial tissue engineering refers to regenerating tissues found in the brain and the face parts such as bone, muscle, and artery. More biocompatible and biodegradable scaffolds are more commensurate with tissue remodeling and more appropriate for cell culture, signaling, and adhesion. Synthetic materials play significant roles and have become more prevalent in medical applications. They have also been used in different forms for producing a microenvironment as ECM for cells. Synthetic scaffolds may be comprised of polymers, bioceramics, or hybrids of natural/synthetic materials. Synthetic scaffolds have produced ECM-like materials that can properly mimic and regulate the tissue microenvironment's physical, mechanical, chemical, and biological properties, manage adherence of biomolecules and adjust the material's degradability. The present review article is focused on synthetic materials used in craniofacial tissue engineering in recent decades.

Antimicrobial Potential of the Green Microalgae Isolated from the Persian Gulf.

Background: We aimed to investigate the antibacterial activity of Persian Gulf microalgae extracts on some Gram-positive and negative bacterial species in order to find new compounds with antibacterial activity. Methods: After sampling microalgae from December 2020 to April 2021 from the northernmost part of Qeshm Island in Persian Gulf, the antibacterial activity of methanolic and ethyl acetate extract of microalgae were tested in three concentrations of 125, 250, and 500 mg/ml on Gram-positive bacteria including Staphylococcus aureus, Bacillus cereus, and Gram-negative bacteria including Pseudomonas aeruginosa and Escherichia coli by disk-diffusion assay and the results were compared with two standard antibiotics including ciprofloxacin and streptomycin. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were assessed spectrophotometrically using microplate and enzyme-linked immunosorbent assay (ELISA) reader. Results: Methanolic and ethyl acetate extracts had antibacterial effects against Gram-positive and negative bacteria. Compared to ethyl acetate extract, the methanolic extract showed stronger effects on both Gram-positive and negative bacteria. The most antibacterial effect was related to methanolic extract with a concentration of 500 mg/ml on S. aureus by 14.6 mm inhibition zone. Evidence from MIC also confirmed that the lowest MIC was belonged to methanolic extract by 0.75 mg/ml against S. aureus. Interestingly, both of these extracts showed more antibacterial activity on Gram-positive bacteria than Gram-negative bacteria. Conclusion: The investigation proved the efficacy of microalgae extracts isolated from Persian Gulf as natural antimicrobials and suggested the possibility of employing them in medicines as antimicrobial agents.

Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae.

Background: Multi-drug resistant (MDR) Klebsiella pneumoniae strains cause the majority of community acquired and life-threatening infections. We aimed to detect the gyrA mutations in the clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumoniae isolated from the patients with urinary tract infections. Methods: Bacterial strains were isolated from the patients with urinary tract infections admitted to a major hospital in Tehran, Iran (2017-2018). Bacterial identification was done according to standard microbiological tests. Antimicrobial susceptibility testing for quinolones and fluoroquinolone antibiotics was done using both disc diffusion and minimal inhibitory concentrations (MICs) methods. PCR-RFLP was used to detect the probable mutation in the gyrA gene in nalidixic acid and ciprofloxacin resistant strains. Finally sequencing was performed to detect point mutations in isolated K. pneumoniae strains. Results: One hundred K. pneumonia isolates were recovered from the urine samples of the clinical cases. Antibiotic resistance testing showed that among all K. pneumoniae isolates, 26% and 19% of the strains were resistant to nalidixic acid and ciprofloxacin respectively. MIC value was ≥4 µg/ml for ciprofloxacin resistant isolates. The results of RFLP on gyrA PCR amplicons using HinfI restriction enzyme showed point mutation in this gene in 46% of nalidixic acid and ciprofloxacin resistant K. pneumonia. The data obtained from the sequencing confirmed the RFLP results and indicated the presence of point mutations in codons 83 and 87 in the gyrA gene which leads to the substitution of different amino acids in gyrA protein. Conclusion: Our findings indicated a relative increased rate of resistance against quinolones and fluoroquinolone antibiotics that raised a concern about extensive dissemination of clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumonia. Point mutation of gyrA gene was responsible for the resistance in our strains however to gain more insight into the molecular characterization of quinolone-resistant isolates, other possible mechanisms of the resistance should also be investigated.