Gene Encoding Research Articles

Cellulomonas taurus sp. nov., a novel bacteria with multiple hydrolase activity isolated from livestock, and potential application in wastewater treatment.

A Gram-positive, smooth, sub-transparent, faint yellow,0.5-0.7µm diameter, rod shaped aerobic or facultative aerobic strain P40-2Twas isolated from livestock farms in Northeast China. Strain P40-2T grew at 25-40°C (optimum 30-38 °C), and in 0-4% (w/v) NaCl (optimum 0%) in LB medium. Based on 16S rRNA gene sequence analysis, strain P40-2T belongs to the class Cellulomonas and is most closely related to C. denverensis strain W6929, C. pakistanensis strain NCCP-11and C. hominis strain CE40.DNA-DNA hybridization rate of strain P40-2T was 29%, and the ANI with C.denverensisstrainW6929 was 85.33%. The genome is 3437431bp long with a G + C content of 71.99%. Of the 3177 predicted genes, 3119 were protein-coding genes and 58 were RNA encoding genes. The chemotaxonomic data: menaquinone was MK-9(H4), anteiso-C15: 0, C16:0 and anteiso-C17: 0 were the major cellular fatty acids, and the main cell-wall amino acids were ornithine,alanine, glycine and glutamate. The cell wall peptidogly can sugars included glucose, rhamnose, galactose and mannose. The polar lipid present were DPG, PG, PE, and PIM. On the basis of DNA-DNA relatedness, phylogenetic position, complete genome sequence and physiological characteristics, strain P40-2T can be differentiated from other species of the genus Cellulomonas with validly published names and thus represents a novel species, for which the name Cellulomonas taurus is proposed. The type strain is Cellulomonas taurus P40-2T (= CGMCC No.1.17732T).The acute toxicity test in mice showed that LD50 of strain P40-2T was rather high with 1.5 × 1011CFU/mouse, which indicated low pathogenicity. Drug susceptibility showed that strainP40-2T was resistant to most antibiotics and only sensitive to six antibiotics. Strain P40-2T contained a variety of hydrolytic enzymes including the ability to hydrolyze cellulose, β-glucan, chitin, xylan, and casein. Microbial flocculant MBF-P40 for sewage was prepared with strain P40-2T, after strain P40-2T was confirmed that had good flocculation effect. MBF-P40 was used to prepare flocculation rate of 99.40%. MBF-P40 treatmented sewage from eight different sources. Flocculation rate for pig farm wastewater was 96.07%, COD removal rate is 71.05%, ammonia nitrogen removal rate is 18.22%. The result shows that MBF-P40 has a good flocculation effect, and good prospect of development and application for wastewater treatment.

ESBL and AmpC β-Lactamase Encoding Genes in E. coli From Pig and Pig Farm Workers in Vietnam and Their Association With Mobile Genetic Elements.

Animals are considered important sources of ESBL/AmpC-producing bacteria in humans. We analyzed indications of transfer of ESBL/AmpC genes between pigs and pig farmers in Vietnam by analyzing whole genome sequences of 114 ESBL/AmpC-producing E. coli isolated from the two hosts, and performed conjugation experiments and plasmid profiling to confirm that such transfer could have happened. ESBL-encoding genes detected in pigs and pig farmers included blaCTX–M-55, blaCTX-M-27, blaCTX-M-65, blaCTX-M-15, blaCTX-M-14, blaCTX-M-3, blaCTX-M-24, and blaCARB-2, and AmpC β-lactamases included blaCMY-2, blaDHA-1, and blaCMY-42. The most frequent ESBL gene, blaCTX-M-55, was carried on plasmid with replicons types IncF, IncX, IncH, IncN, IncR, and IncP. The insertion transposases downstream of the blaCTX-M-55 gene were different in plasmids carried by different strains. The second most detected gene, blaCTX-M-27, is found in a stable genetic arrangement with the same flanking transposons seen across strains, and the gene was located on similar conjugal IncF plasmid types, suggesting a horizontal spread of these plasmids. In three strains, we observed a novel blaCTX-M-27 harboring IncF type of plasmid which had not been reported before. Its closest reference in NCBI was the non-ESBL Salmonella Typhimurium plasmid pB71 that might have experienced an insertion of blaCTX-M-27. Our data also point to an emergence of plasmids co-carrying ESBL genes, mcr genes, quinolones and other antimicrobials resistance determinants, and such plasmids require special attention. Plasmids phylogeny confirmed that the blaCTX-M-55 encoding plasmids varied considerably, while those encoding blaCTX-M-27 were closely related. Plasmids harboring both ESBL genes were confirmed to be conjugative and not to differ in transfer efficacy. The isolates carrying the plasmids, even those with plasmids of similar types, showed wide genetic variation with high number of SNPs, suggesting horizontal spread of plasmids into different clonal lines. Their virulence profiles did not confirm to known pathotypes, suggesting that unrelated commensals are a main reservoir for ESBL and AmpC β-lactamases in both humans and pigs. Overall, despite evidence of transferability of plasmids in the analyzed strains, our findings do not support that ESBL-producing E. coli from pigs or their ESBL/AmpC encoding plasmids are commonly spread to workers in close contact with the animals.

Genetic Identification of Shiga Toxin Encoding Gene from Cases of Multidrug Resistance (MDR) Escherichia coli Isolated from Raw Milk

Escherichia coli is one of bacteria which have resistant to three or more classes of antimicrobial agents. E. coli having resistant to three or more classes of antimicrobial drugs can be defined as multidrug-resistant (MDR) bacteria. The aim of the study was to evaluate the expression of Shiga toxin gen in MDR E. coli. A total of 250 raw milks samples were taken from dairy farms in Kediri, Probolinggo, Pasuruan, Blitar, and Batu Region, East Java Province, Indonesia. Each sample was cultured into enrichment media Brilliant Green Bile Lactose Broth and Eosin Methyllen blue agar, then identified with TSIA agar and IMVIC biochemistry test. Antibiotic sensitivity testing was done using Kirby-Bauer disc diffusion assay on medium Mueller-Hinton agar (Oxoid, CM0337). Antibiotics disks used were 30 µg of Tetracycline (Oxoid, CT0054), 10 µg of Streptomycin (Oxoid, CT0047), 30 µg of Chloramphenicol (Oxoid, CT0013), 5 µg of Trimethoprim (Oxoid, CT0057), and 30 µg of Aztreonam (Oxoid, CT0264). Isolate showing resistance to at least 3 antibiotics disk were then continued with PCR assay to identify Shiga toxin E. coli (STEC) encoding stx2 gene. The study was designed to evaluate the nucleotide analysis of STEC gene. The result showed that 6.25% (1/16) of STEC encoding gene was found in MDR E. coli. This report of molecular identification on the presence of STEC gene in MDR E. coli confirmed a wider spread of MDR E. coli that can threaten animal health and human health.

Polymorphisms in the Gene Encoding Caspase 8 May Predict the Response to First-Line Platinum-Based Chemotherapy in Locally Advanced or Advanced Non-Small-Cell Lung Cancer.

Caspase 8 is a protein involved in the process of cell apoptosis, which may affect the efficacy of anti-cancer treatment. The aim of our study was to determine the impact of polymorphisms in the CASP-8 gene encoding caspase 8 on the prognosis in non-small-cell lung cancer (NSCLC). The study involved 99 patients with newly diagnosed locally advanced or metastatic NSCLC treated with platinum-based chemotherapy. The presence of the GG genotype was associated with distant metastases, smoking, and a family history of cancer. The higher risk of early progression was associated with weight loss and the CASP-8 genotype (GG vs. AG or AA: 20.51% vs. 2.86%). The higher risk of progression-free survival (PFS) shortening was associated with a higher stage of disease (hazard ratio (HR) = 2.50, 95% CI: 1.61–3.89, p < 0.0001), distant metastases (HR = 2.30, 95% CI: 1.42–3.72, p = 0.0016), and the GG genotype (HR = 1.68, 95% CI: 1.10–2.57, p = 0.0152). The influence of the GG genotype on the PFS was confirmed in a multivariate analysis (HR = 1.80, 95% CI: 1.06–3.05, p = 0.0317). We did not confirm the influence of CASP-8 genotypes on the overall survival (OS).

Repurposing of antidiabetics as Serratia marcescens virulence inhibitors.

Serratia marcescens becomes an apparent nosocomial pathogen and causes a variety of infections. S. marcescens possess various virulence factors that are regulated by intercellular communication system quorum sensing (QS). Targeting bacterial virulence is a proposed strategy to overcome bacterial resistance. Sitagliptin anti-QS activity has been demonstrated previously and we aimed in this study to investigate the effects of antidiabetic drugs vildagliptin and metformin compared to sitagliptin on S. marcescens pathogenesis. We assessed the effects of tested drugs in subinhibitory concentrations phenotypically on the virulence factors and genotypically on the virulence encoding genes' expressions. The protection of tested drugs on S. marcescens pathogenesis was performed in vivo. Molecular docking study has been conducted to evaluate the interference capabilities of tested drugs to the SmaR QS receptor. Vildagliptin reduced the expression of virulence encoding genes but did not show in vitro or in vivo anti-virulence activities. Metformin reduced the expression of virulence encoding genes and inhibited bacterial virulence in vitro but did not show in vivo protection. Sitagliptin significantly inhibited virulence factors in vitro, reduced the expression of virulence factors and protected mice from S. marcescens. Docking study revealed that sitagliptin is more active than metformin and fully binds to SmaR receptor, whereas vildagliptin had single interaction to SmaR. The downregulation of virulence genes was not enough to show anti-virulence activities. Hindering of QS receptors may play a crucial role in diminishing bacterial virulence.

The ldhA Gene Encoding Fermentative l-Lactate Dehydrogenase in Corynebacterium Glutamicum Is Positively Regulated by the Global Regulator GlxR

Bacterial metabolism shifts from aerobic respiration to fermentation at the transition from exponential to stationary growth phases in response to limited oxygen availability. Corynebacterium glutamicum, a Gram-positive, facultative aerobic bacterium used for industrial amino acid production, excretes l-lactate, acetate, and succinate as fermentation products. The ldhA gene encoding l-lactate dehydrogenase is solely responsible for l-lactate production. Its expression is repressed at the exponential phase and prominently induced at the transition phase. ldhA is transcriptionally repressed by the sugar-phosphate-responsive regulator SugR and l-lactate-responsive regulator LldR. Although ldhA expression is derepressed even at the exponential phase in the sugR and lldR double deletion mutant, a further increase in its expression is still observed at the stationary phase, implicating the action of additional transcription regulators. In this study, involvement of the cAMP receptor protein-type global regulator GlxR in the regulation of ldhA expression was investigated. The GlxR-binding site found in the ldhA promoter was modified to inhibit or enhance binding of GlxR. The ldhA promoter activity and expression of ldhA were altered in proportion to the binding affinity of GlxR. Similarly, l-lactate production was also affected by the binding site modification. Thus, GlxR was demonstrated to act as a transcriptional activator of ldhA.

Rapid Identification for Serotyping of African Swine Fever Virus Based on the Short Fragment of the EP402R Gene Encoding for CD2-Like Protein

Abstract The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.

Association of Rs13476 Polymorphism in lncRNA GAS5 Encoding Gene with Type 2 Diabetes in Isfahan, Iran

Association of Rs13476 Polymorphism in lncRNA GAS5 Encoding Gene with Type 2 Diabetes in Isfahan, Iran

Time-Course Changes of Extracellular Matrix Encoding Genes Expression Level in the Spinal Cord Following Contusion Injury-A Data-Driven Approach.

The involvement of the extracellular matrix (ECM) in lesion evolution and functional outcome is well recognized in spinal cord injury. Most attention has been dedicated to the “core” area of the lesion and scar formation, while only scattered reports consider ECM modification based on the temporal evolution and the segments adjacent to the lesion. In this study, we investigated the expression profile of 100 genes encoding for ECM proteins at 1, 8 and 45 days post-injury, in the spinal cord segments rostral and caudal to the lesion and in the scar segment, in a rat model. During both the active lesion phases and the lesion stabilization, we observed an asymmetric gene expression induced by the injury, with a higher regulation in the rostral segment of genes involved in ECM remodeling, adhesion and cell migration. Using bioinformatic approaches, the metalloproteases inhibitor Timp1 and the hyaluronan receptor Cd44 emerged as the hub genes at all post-lesion times. Results from the bioinformatic gene expression analysis were then confirmed at protein level by tissue analysis and by cell culture using primary astrocytes. These results indicated that ECM regulation also takes place outside of the lesion area in spinal cord injury.

Exosomes are Secreted at Distinct Densities in Human Cancer Cells and are Promising Vectors for Hydrophobic Drug Delivery

Exosomes are cell-derived vesicles less than 200 nm in size containing gene encoding and gene silencing RNA and cytosolic proteins with roles in intercellular communication. Interest in the use of exosomes as targeted drug delivery vehicles has grown since it was shown that they can bind to specific cells and deliver intact genetic material to the cytosol of target cells. We present the detailed isolation protocol which allowed us to isolate secreted vesicles, including exosomes, from human prostate cancer (PC3) and melanoma (M21) cell lines. Transmission electron microscopy (TEM) showed that the isolated vesicles displayed a cup-shaped morphology as well as prolate shapes previously unreported in the literature. Nanoparticle Tracking Analysis (NTA) and western blot assays demonstrated that these vesicles were largely within the 30 - 200 nm size range expected for exosomes and were enriched for CD63, an exosome marker protein. NTA data further revealed discrete vesicle subpopulations with narrowly defined size ranges and consistent ratios of 30 - 200 nm vesicles to vesicles of all sizes secreted from both cell lines. These findings suggest a cellular control mechanism governing the fraction of secreted vesicles within the 30 - 200 nm range. Our work provides novel insights on exosome biogenesis and highlights the potential advantages of exosomes as hydrophobic drug carriers.

Cross-reactivity of 4CMenB vaccine-induced antibodies against meningococci belonging to non-B serogroups in Italy

ABSTRACT The four-component meningococcal serogroup B vaccine (4CMenB) contains antigens present in the majority of meningococci causing invasive meningococcal disease (IMD) and may potentially offer protection against strains belonging to non-B serogroups. This study aimed to evaluate the ability of 4CMenB-induced antibodies to kill, in a human serum bactericidal assay (hSBA), non-B meningococci belonging to the main genotypes responsible for IMD in Italy. Meningococci, collected between 2015 and 2017, was characterized for PorA, FetA and sequence type, and for clonal complex. Twenty non-B isolates, representative of the most frequent genotypes, were molecularly characterized for 4CMenB antigens and tested in hSBA with sera from 4CMenB-vaccinated infants and adolescents. Among twenty isolates, eleven were serogroup C, five were Y, two W and two X. All isolates contained genes encoding for fHbp and NHBA antigens and four harbored the NadA full-length encoding gene. Positive hSBA titers were obtained against all serogroup W, X and Y isolates and against five serogroup C isolates. These data show that the 4CMenB vaccine can induce bactericidal antibodies against genetically representative meningococcal W, Y and X strains from Italy. For serogroup C, different susceptibilities to killing were observed for strains with similar antigenic repertoires.

Association of YKL-40 Encoding Gene CHI3L1 rs946263 with Insulin Resistance and Severity of Coronary Artery Disease in Type 2 Diabetes Mellitus Patients

Association of YKL-40 Encoding Gene CHI3L1 rs946263 with Insulin Resistance and Severity of Coronary Artery Disease in Type 2 Diabetes Mellitus Patients

The bacterial diversity of raw Moroccon camel milk

Dromedary camel milk is generally considered a valuable and marketable commodity but its production suffers from poor hygienic conditions that result in low microbiological quality and the presence of various pathogens. The objective of the present study was to provide a detailed report of the bacterial species level composition of Moroccan raw camel milk samples that can serve as a starting point for the selection of starter cultures to facilitate a change in manufacturing practices to an improved and safer production system. The composition of the bacterial community in four freshly collected raw camel milk samples was analyzed by performing a large-scale isolation campaign combined with 16S rRNA gene amplicon sequencing. A total of 806 isolates were obtained from four raw camel milk samples using ten combinations of growth media and incubation conditions. Subsequent isolate dereplication using MALDI-TOF mass spectrometry and identification of representative isolates through sequence analysis of protein encoding and 16S rRNA genes revealed the presence of established and novel dairy lactic acid bacteria, as well as bacteria that are considered indicators of poor hygienic conditions and psychrotrophic spoilage organisms. The large numbers of Lactococcus and Enterococcus isolates obtained present an interesting resource for starter culture selection.

UAV Task Allocation for Hierarchical Multiobjective Optimization in Complex Conditions Using Modified NSGA-III with Segmented Encoding

With the recent boom in unmanned aerial vehicle (UAV) technology, many UAV applications involving complex and risky tasks in military and civilian fields have emerged, such as military strikes and disaster monitoring. Task allocation for UAVs is the process of planning the division of work among UAVs, controlled from ground stations by human operators. This study formulates the UAV task-allocation problem as an extended traveling salesman problem and presents a novel UAV task-allocation model for complex air concentration monitoring tasks. Then, an optimized non-dominated sorting genetic algorithm III (NSGA-III) based on a twin-exclusion mechanism, hierarchical objective-domination operator, and segmented gene encoding (i.e., NSGA-III-TEHOD) is developed to solve complex task-allocation problems involving multiple UAVs, hierarchical objectives, obstacles, and ambient wind. The algorithm is tested in several simulations, and the results demonstrate that the new algorithm outperforms NSGA-III, non-dominated sorting genetic algorithm II (NSGA-II), and genetic algorithm (GA) in terms of efficiency of global convergence and early maturation prevention and is available for the hierarchical objective-optimization problems.

Polymorphisms of Encoding Genes IL1RN and P2RX7 in Apical Root Resorption in Patients after Orthodontic Treatment.

External apical root resorption (EARR) is one of the most serious complications associated with orthodontic treatment. The aim of the study was to analyze the relationships between selected single nucleotide polymorphisms (SNPs) in Interleukin 1 receptor antagonist (IL1RN), purinoreceptor P2X7 (P2RX7) and EARR in patients after orthodontic treatment. The study comprised 101 patients who underwent a complex orthodontic treatment with a combination of fixed appliances. Roots were measured based on orthopantomograms and lateral cephalometric radiographs taken before and at the end of the treatment using diagnostic software. Proportional measurements of selected teeth were made using the modified Linge and Linge methods. Based on the presence or absence of EARR, patients were divided into two groups: control group, 61 patients without EARR (with 0.90 ≤ rRCR ≤ 1.00), and EARR group, 40 patients with EARR (rRCR < 0.90). Root resorption in selected groups was also evaluated with the scores of Malmgren and Levander. SNP analysis was performed using the real-time polymerase chain reaction (PCR) method. The analysis indicated that a specific haplotype of P2RX7 (rs208294) and IL1RN (rs419598) modified the risk of EARR development (p < 0.05), with a Bonferroni correction. The analysis of the P2RX7 and IL1RN gene polymorphisms showed that the presence of SNPs of these genes may predispose individuals to EARR. These findings indicate that EARR is a complex condition influenced not only by environmental factors and needs further study on the genetic risk factors.

Is there a correlation between viral load and olfactory & taste dysfunction in COVID-19 patients?

ObjectivesTo evaluate the correlation between cycle threshold (Ct) value and occurrence of olfactory and taste dysfunction in COVID-19 patients. MethodsThis comparative study included COVID-19 patients diagnosed by reverse transcription- polymerase chain reaction (RT-PCR) based test at our hospital with mild to moderate disease. The demographic details and detailed clinical history of the patient, including history of loss of smell and taste was taken at the time of presentation. The patients were divided into 2 groups, group A: COVID-19 patients with OTD; group B: COVID-19 patients without OTD. 100 contiguous patients were recruited in each group. The COVID-19 test by RT-PCR was done and Ct value of the 3 genes: E (Envelope encoding) gene, N (Nucleocapsid encoding) gene, and RdRp (RNA-dependent RNA polymerase) gene, was used for data analysis. The Ct values of each of the three genes were compared between groups A and B. ResultsGroup A and B did not differ significantly in terms of basic demographics. The differences in the Ct values of the 3 genes E gene, N gene and RdRp gene, of group A and B were found to be statistically significant (p = 0.005, p = 0.001 and p = 0.002, respectively). ConclusionThe patients with OTD had a lower Ct value at diagnosis, and hence, a higher viral load than those without OTD. The evaluation of Ct value and viral load in COVID-19 patients may help in further reducing the transmission of the virus in the community.

Impact of COMT val158met on tDCS-induced cognitive enhancement in older adults

BackgroundPrevious studies suggest that genetic polymorphisms and aging modulate inter-individual variability in brain stimulation-induced plasticity. However, the relationship between genetic polymorphisms and behavioral modulation through transcranial direct current stimulation (tDCS) in older adults remains poorly understood. ObjectiveLink individual tDCS responsiveness, operationalized as performance difference between tDCS and sham condition, to common genetic polymorphisms in healthy older adults. Methods106 healthy older participants from five tDCS-studies were re-invited to donate blood for genotyping of apoliproprotein E (APOE: ε4 carriers and ε4 non-carriers), catechol-O-methyltransferase (COMT: val/val, val/met, met/met), brain-derived neurotrophic factor (BDNF: val/val, val/met, met/met) and KIdney/BRAin encoding gene (KIBRA: C/C, C/T, T/T). Studies had assessed cognitive performance during tDCS and sham in cross-over designs. We now asked whether the tDCS responsiveness was related to the four genotypes using a linear regression models. ResultsWe found that tDCS responsiveness was significantly associated with COMT polymorphism; i.e., COMT val carriers (compared to met/met) showed higher tDCS responsiveness. No other significant associations emerged. ConclusionUsing data from five brain stimulation studies conducted in our group, we showed that only individual variation of COMT genotypes modulated behavioral response to tDCS. These findings contribute to the understanding of inherent factors that explain inter-individual variability in functional tDCS effects in older adults, and might help to better stratify participants for future clinical trials.

Association of a Polymorphism in the Gene Encoding Methylenetetrahydrofolate Dehydrogenase 1 (MTHFD1) 1958G&gt;A with Orofacial Cleft

Objective: To evaluate the possible association of a polymorphism in the gene encoding methylenetetrahydrofolate dehydrogenase 1 (MTHFD1), 1958G>A, with the susceptibility to orofacial cleft in an Indonesian population. Material and Methods: A total of 200 stored secondary biological samples from 30 cases of orofacial cleft and 170 unaffected controls were analyzed to determine the polymorphism status at base 1958. The analysis was conducted using the PCR-restriction fragment length polymorphism technique after digestion with the Msp1 restriction enzyme . The samples were then subjected to agarose gel electrophoresis to investigate the presence or absence of the following fragments: genotype GG, 196, 86 and 40 base pairs (bp); genotype AA, 282 and 28 bp and genotype AG, 282, 196, 86, 40 and 28 bp. The test groups were compared using the Chi-square test. Results: The wild-type allele containing 1958G, as well as the genotype GG, were significantly more common in the control group than in the orofacial cleft group. Conclusion: The MTHFD1 1958G>A polymorphism was significantly associated with orofacial cleft susceptibility in the tested Indonesian population.

Detection of Biofilm and Siderophore Encoding Genes Implicated in the Pathogenesis of Klebsiella pneumoniae Isolated from Different Clinical Specimens

Background: Biofilm and siderophores protect K. pneumoniae, making them resistant to antimicrobials and increase the ability to evade immune system. Objective: The aim of the present study was to detect some of the virulence genes responsible for biofilm formation and siderophore production implicated in the pathogenesis of K. pneumoniae. Methodology: Crystal violet method was used to detect biofilm phenotypically, hypermucoviscosity by string test and virulence genes were detected by PCR. Results: Phenotypically, biofilm formation was positive in 76% and negative in 24%. Genotypically, FimH and ycfm were detected in 100% of isolates while mrkD in 98%. Kfu, entB and iutA were found in 100%, 68% and 34% respectively. Phenotypically by string test 66% of isolates were classified as hypermucoviscous, while 34% as classical. rmpA gene was detected in 52% of isolates. Conclusion: FimH, mrkD, ycfm, Kfu and iutA genes are implicated in multisystemic infection of K. pneumoniae. Enterobactin gene is more predominant than other siderophore genes.

Lissencephaly in an epilepsy cohort: Molecular, radiological and clinical aspects.

Lissencephaly is a rare malformation of cortical development due to abnormal transmantle migration resulting in absent or reduced gyration. The lissencephaly spectrum consists of agyria, pachygyria and subcortical band heterotopia. In this study we compared genetic aetiology, neuroradiology, clinical phenotype and response to antiepileptic drugs in patients with epilepsy and lissencephaly spectrum malformations. The study group consisted of 20 patients - 13 males and 7 females, aged 18 months to 21yearsat the time of data collection. Genetic testing was performed by oligonucleotide array comparative genomic hybridization (microarray), multiplex ligation-dependent probe amplification (MLPA), targeted gene panels and whole exome/genome sequencing. All neuroradiological investigations were re-evaluated and the malformations were classified by the same neuroradiologist. Clinical features and response to anti-epileptic drugs (AEDs) were evaluated by retrospective review of medical records. In eleven patients (55%) mutations in PAFAH1B1 (LIS1) or variable microdeletions of 17p13.3 including the PAFAH1B1 gene were detected. Four patients (20%) had tubulin encoding gene mutations (TUBA1A, TUBG1 and TUBGCP6). Mutations in DCX, DYNC1H1, ADGRG1 and WDR62 were identified in single patients. In one patient, a possibly pathogenic intragenic deletion in TRIO was detected. A clear radiologic distinction could be made between tubulinopathies and PAFAH1B1 related lissencephaly. The majority of the patients had therapy resistant epilepsy and epileptic spasms was the most prominent seizure type. The best therapeutic response to seizure control in our cohort was obtained by the ketogenic diet, vigabatrin, clobazam, phenobarbital and valproate. The most common genetic aetiologies in our cohort of 20 individuals with epilepsy and lissencephaly spectrum were intragenic deletions or single nucleotide mutations in PAFAH1B1 or larger deletions in 17p13.3, encompassing PAFAH1B1, followed by mutations in tubulin encoding genes. Radiological findings could reliably predict molecular results only in agyria with a posterior to anterior gradient. Radiological and molecular findings did not correlate consistently with severity of clinical outcome or therapeutic response.