Transcriptomic analysis of human cytomegalovirus to survey the indirect effects on renal transplant recipients.

Post-transplant human cytomegalovirus (HCMV) viremia has been linked to adverse "indirect effects" among transplant patients. HCMV-created immunomodulatory mechanisms could be associated with the indirect effects. OBJECTIVE: In the present study, the RNA-Seq whole transcriptome of renal transplant (RT) patients was analyzed to seek the underlying pathobiologic pathways associated with the long-term indirect effects of HCMV. METHODS: To investigate the activated biological pathways in HCMV infection, total RNA was extracted from PBMCs of 2 RT patients with active HCMV and 2 RT patients without infection and then were sequenced using RNA-Seq. The resulted raw data were analyzed by conventional RNA-Seq software to determine the Differentially Expressed Genes (DEGs). Afterward, Gene Ontology (GO) and pathway enrichment analyses were conducted to determine the enriched pathways and biological processes by DEGs. Eventually, the relative expressions of some significant genes were validated in the twenty external RT patients. RESULT: The analysis of RNA-Seq data related to RT patients with HCMV active viremia led to the identification of 140 up-regulated and 100 down-regulated DEGs. KEGG pathway analysis revealed the enrichment of DEGs in IL18 signaling, AGE-RAGE signaling pathway in diabetic complications, signaling by GPCR, Platelet activation, signaling and aggregation, Estrogen signaling pathway and signaling by Wnt due to HCMV infection. The expression levels of six genes involved in enriched pathways including F3, PTX3, ADRA2B, GNG11, GP9, HBEGF were then verified using RT-qPCR. The results were in consistent with RNA-Seq resultsoutcomes. CONCLUSION: This study specifies some pathobiological pathways which are activated in HCMV active infection and could be linked to the adverse indirect effects caused by HCMV infection in transplant patients.

HIV testing among incarcerated people with a history of HIV-related high-risk behaviours in Iran: Findings from three consecutive national bio-behavioural surveys.

BACKGROUND: Incarcerated people are at a disproportionate risk of contracting HIV. We estimated the prevalence and correlates of HIV testing among incarcerated people with a history of HIV-related high-risk behaviours in Iran. METHODS: Data for this analysis were obtained from three consecutive nationwide bio-behavioural surveillance surveys of a random sample of incarcerated people in 2009 (n = 5953), 2013 (n = 5490), and 2017 (n = 5785). History of testing for HIV in the last 12 months was the primary outcome variable. HIV testing was examined among those with a history of HIV-related high-risk behaviours (i.e., having multiple sex partnerships, injection drug use practices, or a history of having a tattoo). The outcome variable was divided into three categories: Never tested for HIV, ever tested for HIV inside the prison in the last 12 months, and ever tested for HIV outside the prison in the last 12 months. We used multivariable multinomial logistic regression models to examine factors associated with HIV testing. RESULTS: Overall, 8,553 participants with a history of HIV-related high-risk behaviors with valid responses to the HIV testing question were included in the analysis. Although HIV testing inside prison has increased (23% [2009], 21.5% [2013], and 50.3% [2017]: P-value < 0.001), the prevalence of HIV testing outside prison has decreased (7.7% [2009], 7.5% [2013], 4.1% [2017]: P-value < 0.001) over time. Our multivariable multinomial regression model showed older age (Relative-risk ratio [RRR]: 1.24, 95% Confidence Intervals [CI]: 1.05, 1.47), history of the previous incarceration (RRR: 1.46, 95% CI: 1.24, 1.71), currently receiving methadone maintenance therapy inside prison (RRR: 2.09, 95% CI: 1.81, 2.43), having access to condoms inside prison (RRR: 1.42, 95% CI: 1.20, 1.68) and sufficient HIV knowledge (RRR: 1.74, 95% CI: 1.47, 2.05) were significantly associated with an increased probability of having an HIV test in the last 12 months inside prison. CONCLUSION: HIV testing among high-risk Iranian prisoners has increased from 2009 to 2017. However, HIV testing remains considerably low, and half of the incarcerated people with a history of HIV-related high-risk behaviours had never tested for HIV inside prison. Evidence-based programs are needed to optimize HIV testing inside and outside prisons and identify those at greater risk of HIV.

Molecular detection and genotyping of Human Cytomegalovirus in kidney transplant recipients under ganciclovir prophylaxis in Iran.

INTRODUCTION: Human Cytomegalovirus (HCMV) infection is one of the most common viral complications in kidney transplant recipients. Although there are effective treatments strategies for the HCMV infection, this infection is still one of the causes of kidney transplant rejection. METHODS: A total of 246 kidney transplant recipients participated in this cross-sectional study. Viral DNA was extracted from these plasma samples and the presence of HCMV genome was determined by Semi-nested PCR with specific primers for HCMV B glycoprotein gene. Sanger sequencing analyses were carried out to determine HCMV genotypes and Mega x software was used for nucleotide alignment and construction of a phylogenetic tree. RESULTS: Human cytomegalovirus DNA was detected in 11 (4.47%) recipients. According to the phylogenetic analysis, HCMV gB3 was 50% among kidney transplant recipients, followed by gB4 30% and gB1 20% however, the gB2 genotype was not detected. CONCLUSIONS: This study demonstrated that the HCMV infection in our patients is relatively low because all transplant recipients received appropriate prophylaxis, thereby antiviral prophylaxis is recommended for all patients at risk of HCMV infection after kidney transplantation. Also, gB3 was the most predominant genotype among our kidney transplant recipients that was related to the higher rate of prevalence of severe HCMV infections. Moreover, elevated serum creatinine level was detected in patients at the time of detection of HCMV infection.

Proposed Pathway Linking Respiratory Infections with Depression.

Depression is one of the most important causes of disability and loss of useful life of people around the world. Acute respiratory infection caused a large number of severe illnesses and deaths of the world and most of these due to viral infections, which is estimated more than 80% of respiratory infections. Detection of viruses by immune pathogen recognition receptors activates the intracellular signaling cascade and eventually cause produces interferons. Inflammatory process begins with secretion of interferons and the expression of interferon-stimulated genes. One of the most important of these genes is indoleamine-pyrrole 2,3-dioxygenase (IDO), which plays a major role in tryptophan catabolism. IDO is an intracellular monomeric enzyme that is also responsible for breaking down and consuming tryptophan in the Kynurenine pathway. Increased inflammation has been linked to decrease tryptophan concentrations and increase kynurenine levels. We tried to explain the role of inflammation by viral respiratory infections in causing depression.

Vector-borne diseases in Iran: epidemiology and key challenges.

Vector-borne diseases have become a global health concern in recent decades as a result of global warming, globalization, growth in international trade and travel, use of insecticide and drug resistance. This review study addressed the key vector-borne diseases and their current status in Iran to emphasize the requirements for further research on vector-borne diseases. The dispersion patterns of these diseases differ in various regions. Some of them such as Crimean-Congo hemorrhagic fever, and Q fever are distributed all across Iran, whereas some others such as plague, leishmaniasis, tularemia, and malaria are restricted to specific areas. The high prevalence of vectors throughout the country necessitates enhancing the monitoring and surveillance of emerging and reemerging vector-borne diseases and their potential vectors.

First Cases of SARS-CoV-2 in Iran, 2020: Case Series Report.

In Jan 2020, the outbreak of the 2019 novel coronavirus (SARS-CoV-2) in Wuhan, Hubei Province of China spread increasingly to other countries worldwide which WHO declared it as a public health emergency of international concern. Iran was included in the affected countries. Throat swab specimens were collected and tested by using real-time reverse transcription PCR (RT-PCR) kit targeting the E region for screening and RNA dependent RNA polymerase for confirmation. Conventional RT-PCR was conducted for the N region and the PCR products were sequenced by Sanger sequencing. The first seven cases of SARS-CoV-2 infections were identified in Qom, Iran. This report describes the clinical and epidemiological features of the first cases of SARS-CoV-2 confirmed in Iran. Future research should focus on finding the routes of transmission for this virus, including the possibility of transmission from foreign tourists to identify the possible origin of SARS-CoV-2 outbreak in Iran.

Molecular characterization of non-polio enteroviruses isolated from children with acute flaccid paralysis in IRAN, 2015-2018.

In addition to polioviruses, non-polio enteroviruses (NPEVs) are frequently isolated from patients with acute flaccid paralysis (AFP) worldwide. In polio-free countries, there have been expectations that with disappearing wild poliovirus from the community, the rate of AFP would decrease, but the increasing number of AFP cases proved this notion to be wrong. There are speculations that NPEVs might be the cause of increasing AFP rate. The aim of this study was to investigate frequency, genetic diversity, circulation patterns of NPEVs isolated from AFP cases in Iran from 2015 to 2018. Fifty-three NPEVs were isolated from stool specimens of AFP cases during four years of AFP surveillance. Nested PCR and VP1 sequencing revealed 20 NPEV types in which Echovirus 3 (13.2%), Echovirus 6 (13.2%), Echovirus 7 (7.5%), Echovirus 13 (7.5%) and Echovirus 21 (7.5%) were the most frequent. Coxsackie B viruses were isolated for the first time in AFP cases in Iran. The phylogenetic analysis of Echovirus 3 and Echovirus 6 revealed that Iranian echovirus strains belonged to the same cluster, indicating these viruses have been circulating in Iran for a long time. Compared to global Echovirus 3 and Echovirus 6 references, Echovirus 3 and Echovirus 6 strains detected in this study were closely related to Indian and Malaysia strains, respectively. The results of this study demonstrated a wide variety of NPEV types in Iranian patients, some of which had not been reported in previous studies. Moreover, this study highlights the need for NPEV surveillance in AFP cases.

Enteroviruses and Adenoviruses in stool specimens of paralytic children- can they be the cause of paralysis?

BACKGROUND AND OBJECTIVES: Acute flaccid paralysis (AFP) is a complicated clinical syndrome with a wide range of potential etiologies. Several infectious agents including different virus families have been isolated from AFP cases. In most surveys, Non-polio Enteroviruses (NPEVs) have been detected as main infectious agents in AFP cases; however, there are also some reports about Adenovirus isolation in these patients. In this study, NPEVs and Adenoviruses in stool specimens of AFP cases with or without Residual Paralysis (RP) with negative results for poliovirus are investigated. MATERIALS AND METHODS: Nucleic acid extractions from 55 AFP cases were examined by nested PCR or semi-nested PCR with specific primers to identify NPEVs or Adenoviruses, respectively. VP1 (for Enteroviruses) and hexon (for Adenoviruses) gene amplification products were sequenced and compared with available sequences in the GenBank. RESULTS: From 55 fecal (37 RP+ and 18 RP-) specimens, 7 NPEVs (12.7%) (2 cases in RP+) and 7 Adenoviruses (12.7%) (4 cases in RP+) were identified. Echovirus types 3, 17 and 30, Coxsackie virus A8, and Enterovirus 80 were among NPEVs and Adenoviruses type 2 and 41 were also identified. CONCLUSION: Our finding shows that NPEVs and Adenoviruses may be isolated from the acute flaccid paralyses but there is no association between the residual paralyses and virus detection.

The frequency of Klebsiella pneumonia encoding genes for CTX-M, TEM-1 and SHV-1 extended-spectrum beta lactamases enzymes isolated from urinary tract infection.

BACKGROUND: The extended- spectrum ß-lactamase producing bacteria are widely spread worldwide. The productions of these enzymes cause bacterial resistance to a wide range of antibiotics. The aim of this study was to investigated the frequency of K. pneumonia encoding genes for CTX-M, TEM-1 and SHV-1 extended-spectrum beta lactamases enzymes isolated from urinary tract infection. METHODS: This study is cross-sectional study. All K. pneumonia isolates from urine samples, which had grown on media culture more than 105 were delivered to the medical microbiology laboratory. K. pneumonia susceptibility of 198 samples were confirmed by disk diffusion. The gene frequency of genes was determined using PCR, and analyzed using SPSS version 21 software. FINDING: Most of the K. pneumonia isolated from urine producing ß-lactamase were resistant to cotrimoxazole (53.2%) followed by cefotaxime (50%), ceftazidime, ceftriaxone (40.3%), nalidixic acid (17.8%), amikacin and imipenem (1.6%) and meropenem (0%) respectively. Out of the 198 confirmed isolates of K. pneumonia, 62 cases (31.3%) have the gene phenotype of broad spectrum ß-lactamase enzymes and highest frequency of gene phenotype was related to the SHV-1 gene (85.5%). Then in the terms of abundance from highest to lowest CTXM-3 (56.5%), CTXM-1 (27.4%), TEM-1 (16.1%) and CTXM-2 (8.1%), were respectively. CONCLUSION: This study showed that K. pneumonia isolated from urine producing ß-lactamase were resistance to a wide range of antibiotics. Due to the increasing resistance of most antibiotics, control and supervision in the use of antibiotics and identification of broad spectrum ß-lactamase enzymes by phenotypic methods appears to be essential.

Emerging and re-emerging infectious diseases in Iran.

Despite development of preventive and controlling strategies regarding infectious diseases, they are still considered as one of the most significant leading causes of morbidity and mortality, worldwide. Changes in humans' demographics and behaviors, microbial and ecological alterations, agricultural development, international travels and susceptibility to infectious diseases have resulted in increased reports of emerging infectious diseases (EIDs) and reemerging infectious diseases (RIDs) in various geographical areas. Because of the various types of geographic properties in Iran, substantial climatic variability, as well as unstable political situations and poor public health conditions in some of neighboring countries, EIDs and RIDs are serious public health problems; among them, zoonotic and drug resistant diseases are the most significant. Hence, this review provides an overview of the significant bacterial, viral and fungal EIDs and RIDs in Iran regarding their epidemiological aspects.

Molecular characterization of Staphylococcus aureus isolates from southwest of Iran using spa and SCCmec typing methods.

Staphylococcus aureus remains a major cause of nosocomial infection worldwide. Characterization of S. aureus isolates circulating in the southwest of Iran will contribute to understand and control the spread of the strains in this area. spa and SCCmec typing methods were used for genotyping of 125 S. aureus isolates obtained from two teaching hospitals in Ahvaz. Drug susceptibility testing was performed by using disk diffusion method. Frequency of the methicillin resistant S. aureus (MRSA) isolates was 39% (n = 34) and 27% (n = 10) in Emam Khomeini and Golestan hospitals, respectively. Except for Erythromycin, MRSA strains showed high rate of resistance to antimicrobial agents including penicillin (100%), norfloxacine (80%), azitromycin (80%), ciprofloxacin (80%), gentamycin (77%), cotrimoxazole (75%), cephotaxime. All isolates were sensitive to vancomycin. Out of 44 MRSA strains, 39 (88.5%) were SCCmec III, three (7%) were IVc and two (4.5%) of them were nontypeable. spa types t037 (26 isolates; 59%), and t1149 (25 isolates; 31%) were the most dominant types found in MRSA and methicillin sensitive S. aureus (MSSA) strains, respectively. We found SCCmec type III as the most prominent type indicating that most of the studied bacterial population had hospital origin. spa type t037, the most frequent genotype in this study were significantly (100%) associated with MRSA. For the first time we are reporting spa types t692, t706 and t018 from Iran and t342, t704, t2622, t5598, t11270 and t2864 from Asia. Moreover we are reporting types t6871 and t2684 for the second time in the world.

High Frequency of Class 1 Integrons in Escherichia coli Isolated From Patients With Urinary Tract Infections in Yasuj, Iran.

BACKGROUND: Most urinary tract infections (UTI) are caused by Escherichia coli. Integrons have an important role in distributing antibiotic resistance genes among bacteria. OBJECTIVES: The aim of this study was to investigate the presence of class 1, 2 and 3 integrons and their association with antibiotic resistance in E. coli isolated from patient with UTI in Yasuj, Iran. PATIENTS AND METHODS: In this cross-sectional study a total of 200 E. coli were collected from 1820 patients diagnosed with UTI that had been referred to two clinical laboratories between February 2013 and November 2014 in Yasuj city, southwest of Iran. Susceptibility of isolates to 11 different antibiotics was determined by the disk agar diffusion method. multiplex-polymerase chain reaction (PCR) was used for detection of class 1, 2 and 3 integrons. The data were analyzed using the SPSS software (version 16) and the chi-square test. A P value of < 0.05 was considered statistically significant. RESULTS: The highest rate of resistance was observed toward cephalothin (99%) and amoxicillin (76%) while only two (1%) isolates showed resistance to imipenem. Overall, 79% of isolates were multi drug resistant (MDR). Class 1 and 2 integrons were detected in 104 (52%) and 5 (2.5%) isolates respectively, while none of the isolates were positive for class 3 integrons. A significant association was observed between the presence of integrons and resistance to co-trimoxazole, nalidixic acid, ciprofloxacin, amoxicillin, ceftazidime and tetracycline (P < 0.05). CONCLUSIONS: High MDR isolates of E. coli were observed in this study. The significant association between class 1 integrons and resistance to ciprofloxacin, nalidixic acid, co-trimoxazole, amoxicillin, ceftazidime and tetracycline showed that class 1 integrons have an important role in resistance to these antibiotics in this region.

High frequency of multidrug-resistant Staphylococcus aureus with SCCmec type III and Spa types t037 and t631 isolated from burn patients in southwest of Iran.

Methicilin resistance Staphylococcus aureus (MRSA) infections are the major challenges in hospitals, especially in the burn units. The use of molecular typing methods is essential for tracking the spread of S. aureus infection and epidemiological investigations. The aim of this study was to find the profile of the spa types and also the prevalence of each SCCmec type of S. aureus strains in a central burn hospital in southwest of Iran. A total of 81 non-duplicate S. aureus were isolated from burn patients between April 2011 and February 2012. The susceptibility of the isolates against 13 different antibiotics was tested by disk agar diffusion (DAD) method. MRSA strains were identified by amplification of mecA gene. Multiplex-polymerase chain reaction (PCR) technique was used to determine the SCCmec types of MRSA strains and all the S. aureus isolates were typed by spa typing method. Detection of mecA gene showed that 70 (86.4%) of the isolates were MRSA. The highest rate of resistance was observed for penicillin (97.5%) and erythromycin (77.8%). None of the isolates were resistant to vancomycin. Sixty-seven of the 70 MRSA isolates harbored only SCCmec type III and three untypeable isolates. Five different spa types were detected. The most common spa types were t037 (42.5%) and t631 (34.5%) and were only found in MRSA isolates. Only SCCmec type III was found in burn patients which emphasizes the HA-MRSA origin of these strains. Only five different spa types identified in this study are in accordance with one SCCmec type which indicates that a limited number of bacterial colons are circulated in the burn unit in this hospital.

Spread of extended-spectrum ß-lactamase genes of blaOXA-10, blaPER-1 and blaCTX-M in Pseudomonas aeruginosa strains isolated from burn patients.

BACKGROUND: Pseudomonas aeruginosa is resistant to many antibiotics due to production of different classes of extended spectrum ß-lactamases (ESBLs). Prevalence of ESBLs among P. aeruginosa has been increased in recent years, demonstrate a serious health problem especially in burn units worldwide. OBJECTIVE: Present study was designed to determine the ESBL producing strains and identify the genes encoding three different ESBLs of bla PER-1, bla OXA-10 and bla CTX-M genes in P. aeruginosa isolates from burn patients. METHODS: In total 185 clinical isolates of P. aeruginosa were collected from infectious wounds of hospitalized burn patients. Antimicrobial susceptibility testing and phenotypic detection of ESBL were performed by disk diffusion method and Double disk Synergy Test (DDST). Polymerase Chain Reaction (PCR) was done for detection of bla OXA-10, bla PER-1 and bla CTX-M ESBL encoding genes. RESULTS: In total, 176 (95.13%) isolates were multidrug resistant. The DDST demonstrated 96 (51.9%) isolates as putative ESBL producers with 100% or highly resistance to ofloxacin, cephalexin, aztreonam (97.57%) and ceftriaxone (91.6%). By PCR amplification, bla PER-1, bla OXA-10 and bla CTX-M genes were detected in 52 (54.16%), 66 (68.75%) and 1 (1.04%) isolates of ESBL producers respectively. Forty-three isolates (44.79%) were simultaneously positive for both bla OXA-10 and bla PER-1 related genes. CONCLUSION: The rate of ESBL producing P. aeruginosa was notable in present study. Since there are only limited effective antibiotics against the bacterium, therefore all isolates must be investigated by antimicrobial susceptibility testing, which limits resistance development in burn units and helps the management of treatment strategy.

Bacterial urinary tract infection in renal transplant recipients and their antibiotic resistance pattern: A four-year study.

BACKGROUND AND OBJECTIVE: Urinary tract infections (UTIs) are the most common infections in renal transplant recipients and are considered a potential cause of bacteremia, sepsis, and affects graft outcomes. The aim of the present study was to determine the incidence of UTI among renal transplant recipients and investigation of antimicrobial susceptibility pattern of causative agents. MATERIALS AND METHODS: In total, 1165 patients from March 2009 to December 2012, in transplant center of Golestan Hospital, Ahvaz, Iran, were investigated. Qualitative urine cultures were performed for all cases, causative microorganisms were identified and colony count was performed according to the standard protocol. Antibiotic susceptibility testing was then performed to determine the susceptibility pattern of recovered bacteria from confirmed UTIs. RESULTS: UTI was diagnosed in 391 patients(33.56%). Gram-negative bacteria were the most prevalent isolated microorganisms with E. coli (43.53%), followed by Enterobacter spp. (35.37%) as the major organisms. Among Gram positives, Coagulase-negative Staphylococci was isolated from 6.8% of cases. The rate of resistance to all tested antibiotics was highest in Enterobacter spp., however the most common resistance were seen against cefixime, cephalotin, and cotrimoxazole in all tested gram negatives. CONCLUSION: the rate of UTIs among renal transplant recipients was noticeable in this study with high antibiotic resistance. Multi-resistant bacterial infections are potentially life-threatening emerging problem in renal transplantation. Prophylactic measures must be applied to patients at greater risk.

Genotyping of Pseudomonas aeruginosa strains isolated from burn patients by RAPD-PCR.

BACKGROUND: Pseudomonas aeruginosa is one of the important causes of nosocomial infections that easily gains resistance to many antibiotics. This opportunistic pathogen is a major health hazard particularly in immunodeficient patients, patients in intensive care units (ICU) and burn units with life threatening outcome. The bacterium may be originated from different or common sources, and comprises a high colonization and transmission capacity. OBJECTIVE: The aim of present study was to investigate the genotypic variation of Pseudomonas aeroginosa strains isolated from burn patients by using Random Amplified Polymorphic DNA (RAPD) method. METHODS: Totally 70 clinical samples were collected from burn patients in Taleghani Burn Hospital of Ahvaz. Fifty out of total samples were positive for P. aeruginosa by application of conventional culture and biochemical identification tests. DNA was extracted from the isolates and the RAPD-PCR method was applied to the DNA extracts according to standard method using a short single primer of 272. The technique created repetitive electrophoresis patterns which was used for genotypic differentiation. RESULTS: RAPD-PCR, created 9 genotypic profiles designated as I-IX with base pair length ranging from 180 to 2700. Each genotype showed between 3 and 6 different weight DNA bands. Genotype I was the most prevalent, identified in 10 bacterial isolates (20%). Genotypes I, II and VI were mostly common in patients with more severe burn, and were mainly isolated from wound and blood samples obtained from the same patients. CONCLUSION: In present study, we found RAPD-PCR technique as a useful tool for investigation of the genetic variation among P. aeruginosa strains. This is a rapid, low cost, genotypic method with high discriminatory power. The results could assist to screen for the original of infection caused by this organism with subsequent control of colonization and transmission.