Evaluation of Phenotypic Tests to Detect Extended-Spectrum ß-Lactamase (ESBL)-Producing Klebsiella oxytoca Complex Strains.

Klebsiella oxytoca complex (KoC) species may overproduce their chromosomal class A OXY ß-lactamases, conferring reduced susceptibility to piperacillin-tazobactam, expanded-spectrum cephalosporins and aztreonam. Moreover, since clavulanate maintains its ability to inhibit these enzymes, the resulting resistance phenotype may falsely resemble the production of acquired extended-spectrum ß-lactamases (ESBLs). In this work, a collection of 44 KoC strains of human and animal origin was characterized with whole-genome sequencing (WGS) and broth microdilution (BMD) susceptibility testing. Comparison of ESBL producers (n = 11; including CTX-M-15 [n = 6] and CTX-M-1 [n = 5] producers) and hyperproducers of OXYs (n = 21) showed certain phenotypic differences: piperacillin-tazobactam (MIC90s: 16 versus >64 µg/mL), cefotaxime (MIC90s: 64 versus 4 µg/mL), ceftazidime (MIC90s: 32 versus 4 µg/mL), cefepime (MIC90s: 8 versus 4 µg/mL) and associated resistance to non-ß-lactams (e.g., trimethoprim-sulfamethoxazole: 90.9% versus 14.3%, respectively). However, a clear phenotype-based distinction between the two groups was difficult. Therefore, we evaluated 10 different inhibitor-based confirmatory tests to allow such categorization. All tests showed a sensitivity of 100%. However, only combination disk tests (CDTs) with cefepime/cefepime-clavulanate and ceftazidime/ceftazidime-clavulanate or the double-disk synergy test (DDST) showed high specificity (100%, 95.5%, and 100%, respectively). All confirmatory tests in BMD or using the MIC gradient strip did not perform well (specificity, ≤87.5%). Of note, ceftazidime/ceftazidime-avibactam tests also exhibited low specificity (CDT, 87.5%; MIC gradient strip, 77.8%). Our results indicate that standard antimicrobial susceptibility profiles can raise some suspicion, but only the use of cefepime/cefepime-clavulanate CDT or DDST can guarantee distinction between ESBL-producing KoC strains and those hyperproducing OXY enzymes.

Practical Agar-Based Disk Diffusion Tests Using Sulfamoyl Heteroarylcarboxylic Acids for Identification of Subclass B1 Metallo-ß-Lactamase-Producing Enterobacterales.

The worldwide distribution of carbapenemase-producing Enterobacterales (CPE) is a serious public health concern as they exhibit carbapenem resistance, thus limiting the choice of antimicrobials for treating CPE infections. Combination treatment with a ß-lactam and one of the newly approved ß-lactamase inhibitors, such as avibactam, relebactam, or vaborbactam, provides a valuable tool to cope with CPE; however, these inhibitors are active only against serine-type carbapenemases and not against metallo-ß-lactamases (MßLs). Therefore, it is important to readily differentiate carbapenemases produced by CPE by using simple and reliable methods in order to choose an appropriate treatment. Here, we developed three practical agar-based disk diffusion tests (double-disk synergy test [DDST], disk potentiation test, and modified carbapenem inactivation method [mCIM]) to discriminate the production of subclass B1 MßLs, such as IMP-, NDM-, and VIM-type MßLs, from the other carbapenemases, especially serine-type carbapenemases. This was accomplished using B1 MßL-specific sulfamoyl heteroarylcarboxylic acid inhibitors, 2,5-dimethyl-4-sulfamoylfuran-3-carboxylic acid (SFC) and 2,5-diethyl-1-methyl-4-sulfamoylpyrrole-3-carboxylic acid (SPC), originally developed by us. The DDST and mCIM using SFC and SPC revealed high sensitivity (95.3%) and specificity (100%) in detecting B1 MßL-producing Enterobacterales. In the disk potentiation test, the sensitivities using SFC and SPC were 89.1% and 93.8%, respectively, whereas the specificities for both were 100%. These methods are simple and inexpensive and have a high accuracy rate. These methods would therefore be of immense assistance in the specific detection and discrimination of B1 MßL-producing Enterobacterales in clinical microbiology laboratories and would lead to better prevention against infection with such multidrug-resistant bacteria in clinical settings.

Antibiogram of ESBL and MBL producing Pseudomonas aeruginosa among the population of Hazara division, KPK, Pakistan.

OBJECTIVE: To investigate the frequency rate and sensitivity pattern of extended-spectrum beta-lactamase and metallobeta- lactamase producing Pseudomonas aeruginosa isolated from major hospitals. METHODS: The cross-sectional study was conducted in the Microbiology section of the Pathology Department of Ayub Medical College, Abbottabad, Pakistan, from September 2017 to April 2018, and comprised clinical samples collected from different medical wards of major hospitals in the study area. For the selective growth of Pseudomonas aeruginosa, Cetrimide agar was used, and different antibiotics were evaluated for the sensitivity pattern following Kirby-Bauer diffusion method. Pseudomonas aeruginosa producing extended-spectrum beta lactamase and metallo-beta-lactamase were identified through double disk synergy test and imipenem ethylenediaminetetraacetic acid tests respectively. Patient's demographic and medical history was noted on a proforma. Data was analysed using SPSS 22.0. RESULTS: Of the 242 samples screened, 46 (19%) were positive for Pseudomonas aeruginosa. These samples were highly sensitive to levofloxacin, amikacin, imipenem, meropenem and ciprofloxacin (p<0.05). Of the positive cases, 11 (23.91%) were detected for extended-spectrum beta-lactamase production, while 3 (6.52%) samples were detected for metallo-beta-lactamase production. CONCLUSIONS: Pseudomonas aeruginosa samples were widely resistant to most antibiotics, but were sensitive for some antibiotics which may be recommended by physicians when treating Pseudomonas aeruginosa infection.

Body composition during early infancy and developmental progression from 1 to 5 years of age: the Infant Anthropometry and Body Composition (iABC) cohort study among Ethiopian children.

Early nutrition and growth have been found to be important early exposures for later development. Studies of crude growth in terms of weight and length/height, however, cannot elucidate how body composition (BC) might mediate associations between nutrition and later development. In this study, we aimed to examine the relation between fat mass (FM) or fat-free mass (FFM) tissues at birth and their accretion during early infancy, and later developmental progression. In a birth cohort from Ethiopia, 455 children who have BC measurement at birth and 416 who have standardised rate of BC growth during infancy were followed up for outcome variable, and were included in the statistical analysis. The study sample was restricted to mothers living in Jimma town who gave birth to a term baby with a birth weight ≥1500 g and no evident congenital anomalies. The relationship between the exposure and outcome variables was examined using linear-mixed regression model. The finding revealed that FFM at birth was positively associated with global developmental progression from 1 to 5 years (ß=1·75; 95 % CI 0·11, 3·39) and from 4 to 5 years (ß=1·34; 95 % CI 0·23, 2·44) in the adjusted model. Furthermore, the rate of postnatal FFM tissue accretion was positively associated with development at 1 year of age (ß=0·50; 95 % CI 0·01, 0·99). Neither fetal nor postnatal FM showed a significant association. In conclusion, fetal, rather than postnatal, FFM tissue accretion was associated with developmental progression. Intervention studies are needed to assess whether nutrition interventions increasing FFM also increase cognitive development.

Double-Layered Hollow Mesoporous Cuprous Oxide Nanoparticles for Double Drug Sequential Therapy of Tumors.

Cancer stem cells (CSCs) are one of the determinants of tumor heterogeneity and are characterized by self-renewal, high tumorigenicity, invasiveness, and resistance to various therapies. To overcome the resistance of traditional tumor therapies resulting from CSCs, a strategy of double drug sequential therapy (DDST) for CSC-enriched tumors is proposed in this study and is realized utilizing the developed double-layered hollow mesoporous cuprous oxide nanoparticles (DL-HMCONs). The high drug-loading contents of camptothecin (CPT) and all-trans retinoic acid (ATRA) demonstrate that the DL-HMCON can be used as a generic drug delivery system. ATRA and CPT can be sequentially loaded in and released from CPT3@ATRA3@DL-HMCON@HA. The DDST mechanisms of CPT3@ATRA3@DL-HMCON@HA for CSC-containing tumors are demonstrated as follows: 1) the first release of ATRA from the outer layer induces differentiation from CSCs with high drug resistance to non-CSCs with low drug resistance; 2) the second release of CPT from the inner layer causes apoptosis of non-CSCs; and 3) the third release of Cu+ from DL-HMCON itself triggers the Fenton-like reaction and glutathione depletion, resulting in ferroptosis of non-CSCs. This CPT3@ATRA3@DL-HMCON@HA is verified to possess high DDST efficacy for CSC-enriched tumors with high biosafety.

Detection of extended-spectrum ß-lactamases in Klebsiella pneumoniae: comparison of phenotypic characterization methods.

OBJECTIVE: Extended-spectrum ß-lactamase producing K. pneumoniae is a serious threat to the patients. This manuscript shows the comparison of phenotypic characterization methods used for ESBL K. pneumoniae and frequency distribution of these isolates in various clinical samples. METHODOLOGY: Eleven different types of pathological samples collected on various time intervals were analyzed. K. pneumoniae were identified with API 20E system (bioMerieux) and initial screening of ESBL K. pneumoniae was performed using the ceftazidime antimicrobial disc. Double-disc synergy test (DDST) and CLSI confirmatory test were compared for the phenotypic detection of ESBL K. pneumoniae. RESULTS: A total number of 214 ESBL producing K. pneumoniae were isolated from various clinical samples. Frequency distribution of ESBL producing K. pneumoniae was found to be highest among blood 117 (54.7%) and urine 46 (21.5%) samples. Data regarding the use of various interventions among these patients showed most common presence of intravenous line 209 (97.7%) and urinary catheters 46 (21.5%). Comparison of DDST and CLSI confirmatory test showed that the DDST detected 145 (67.8%) isolates while 213 (99.5%) ESBL K. pneumoniae were characterized by CLSI confirmatory test. CONCLUSION: The use of CLSI confirmatory test is very efficient in the early detection of ESBL K. pneumoniae especially when the facilities for molecular characterization are not available.

Phenotypic characterization of ESBL producing Enterobacter cloacae among children.

OBJECTIVE: The emergence of ESBL producing Enterobacter cloacae in clinical isolates is posing a serious threat for treating nosocomial infections. The aim of the study was to determine the frequency of extended spectrum ß-lactamase (ESBL) producing Enterobacter cloacae and to compare the phenotypic methods used for the characterization of ESBL producing strains. METHODOLOGY: This cross sectional observational study was conducted during April 2011 to March 2012 at Microbiology department of The Children's Hospital and Institute of Child Health, Lahore. A total number of 20,257 various clinical samples were analyzed during the study period. Enterobacter cloacae were identified using API 20E system and ESBL detection was carried out using double-disk synergy test (DDST) and CLSI confirmatory test. RESULTS: Enterobacter cloacae were isolated from 221 samples, out of which 33 (14.93%) were ESBL producers and 188 (85.07%) were non-ESBL producers. The gender distribution of ESBL producing Enterobacter cloacae was 21 (63.6%) in males and 12 (36.4%) in females. Highest frequency (63%) of ESBL producing Enterobacter cloacae was detected in blood samples. Comparison of DDST and CLSI confirmatory test showed that 25 (75.75%) isolates were characterized by DDST and 33 (100%) using CLSI confirmatory test. CONCLUSION: The present study shows moderately high frequency of ESBL producing Enterobacter cloacae among children. DDST was found to be less efficient in ESBL detection as compared to CLSI confirmatory test.

Comparative evaluation of phenotypic and genotypic methods for the rapid and cost-effective detection of carbapenemases in extensively drug resistant Klebsiella pneumoniae.

PURPOSE: Carbapenemases are the enzymes that can hydrolyze carbapenems and other ß-lactam antibiotics. These enzymes confer resistance to multiple antibiotics and act as a stumbling block in the treatment of infections caused by gram-negative bacteria. Therefore, rapid and specific detection of these enzymes is crucial for deciding the course of treatment and better clinical outcomes. MATERIAL AND METHODS: This study was conducted to compare various phenotypic and PCR based methods for the detection of carbapenemases in carbapenem- and colistin-resistant Klebsiella pneumoniae. One hundred clinical isolates of extensively resistant Klebsiella pneumoniae were included in the study. Phenotypic detection for carbapenemases was performed by Rapidec® Carba NP (Biomerieux), modified carbapenem inactivation method (mCIM), imipenem-ethylenediaminetetraacetic acid disk synergy (EDS), double disk synergy test using mercaptopropionic acid (DDST-MPA), and combined disk method (CD) and for colistin by microbroth dilution method. Genotypic detection for carbapenemases and colistin resistance was performed by targeted PCR. RESULTS: The sensitivity of Carba NP test and mCIM were positive in 95% and 96% respectively and specificity was 100% for both methods. The sensitivity of EDS, DDST-MPA, and CD were 55.6%, 88.9% and 54.5% respectively. Among the carbapenem resistance genes, blaOXA-48 (82%) genes were the most prevalent. Among metallo-beta lactamases, blaVIM (56%) was most common followed by blaNDM (54%) and blaIMP (20%). The mcr-1 gene for colistin resistance was not detected in any isolate. CONCLUSION: Among the five phenotypic assays analyzed, the mCIM is the most simple, inexpensive, accurate and reproducible method for carbapenemase detection in Klebsiella pneumoniae. The DDST-MPA test provides the best sensitivity for the detection of carbapenemases, although specificity is low. These tests, when applied in a clinical laboratory and assessed by the microbiologist, can help in guiding the course of treatment.

Developmental outcomes in children from cryopreserved embryos compared to their siblings from fresh embryo transfers: a case series.

OBJECTIVE: This study aimed to evaluate the developmental outcomes in children from cryopreserved embryos, with minimum influences of interparental variation that would cause potential bias. Hence we examined siblings, in which the older sibs were from fresh embryo transfers, while the younger sibs were from cryopreserved embryos. METHODS: Three pairs of siblings were evaluated. All routine prenatal and neonatal evaluation were performed, while personal-social, language, fine and gross motor evaluation were all evaluated by the Denver Developmental Screening Test (DDST)-II. Wechsler Preschool and Primary Scale of Intelligence (WPPSI) test was used to measure the Intelligent Quotient (IQ) in 5 of 6 children. RESULTS: Standard prenatal measurements of all children suggested uneventful pregnancies, followed by uneventful deliveries. DDST-II results showed that the aspects of personal-social, language, fine and gross motor in every child are as expected according to their ages. Results from WPPSI tests suggest that 5 of 6 evaluated children acquired average to high-average intelligences. CONCLUSIONS: The results suggest that the developmental outcomes in children from cryopreserved embryos have no significant differences with the outcomes in children from fresh embryo transfers.

Developmental evaluation in children experiencing febrile convulsions.

BACKGROUND: The objective of this study was to determine the effect of febrile convulsion (FC) on neuromotor development. METHODS: Data of 325 patients, who were followed up at our outpatient clinic and diagnosed with FC between January 2012 and December 2018, were retrospectively evaluated. Of these patients, 203 underwent the Denver Developmental Screening Test II (DDST II) and were included in the study as the patient group and 100 healthy children as the control group. RESULTS: Of the study group, 84 (41.4%) were girls and 119 (58.6%) were boys (B/G: 1.4). Of all patients, 163 (80.3%) were diagnosed with simple FC, 22 (10.8%) with complicated FC, and 18 (8.9%) with FC+. There was no significant relationship found between FC subtypes and gender, family history of FC, family history of epilepsy, iron (Fe) deficiency, and Fe deficiency anemia. DDST II subtest points were significantly lower in all developmental areas in the patient group when compared to the controls (p < 0.001), while suspected and abnormal test results were higher in all developmental areas in the patient group compared to the controls (p=0.01). It was also determined that the language points were lower as the age of first seizure increased (r=- 0.319, p < 0.01). CONCLUSIONS: Although FC is known to usually having a good prognosis, the low DDST II test results measured in this study indicated that the FC may pose a developmental risk and patients with FC should be followed up in terms of developmental features. Because of the retrospective nature of the study, there was no `preconvulsion` developmental evaluation. This is a major limitation of our study.

Neurodevelopmental Findings and Epilepsy in Malformations of Cortical Development.

AIM: The purpose of this study is to classify the malformations of cortical development in children according to the embryological formation, localization, and neurodevelopmental findings. Seizure/epilepsy and electrophysiological findings have also been compared. MATERIAL AND METHODS: Seventy-five children (age: 1 month-16.5 years; 56% male) followed with the diagnosis of malformation of cortical development, in Marmara University Pendik Research and Educational Hospital Department of Pediatric Neurology, were included in the study. Their epilepsy characteristics, electroencephalogram (EEG) findings, and prognosis were reported. Neurodevelopmental characteristics were evaluated by the Bayley Scales of Infant and Toddler Development (Bayley-III) for the ages of 0-42 months (n = 30); the Denver Developmental Screening Test-II (DDST-II) for ages 42 months-6 years (n = 11); and the Wechsler Intelligence Scales for Children (WISC-R), used for children 6 years and older (n = 34). RESULTS: The patients were classified as 44% premigrational (14.6% microcephaly, 24% tuberous sclerosis, 2.7% focal cortical dysplasia, 1.3% hemimegalencephaly, and 1.3% diffuse cortical dysgenesis); 17.3% migrational (14.6% lissencephaly, 2.7% heterotopia); and 38.6% postmigrational (14.6% schizencephaly, 24% polymicrogyria) developmentally. According to involved area, the classification was 34.7% hemispheric/multilobar, 33.3% diffuse, and 32% focal. Seventy-five percent of the patients had a history of epilepsy, and 92% were resistant to treatment. The seizures started before the age of 12 months in diffuse malformations, and epileptic encephalopathy was more common in microcephaly with a rate of 80% and lissencephaly with a rate of 54.5% in the first EEGs. Ninety-five percent of patients had at least one level of neurodevelopmental delay detected by DDST/Bayley-III; this was more common in patients with accompanying epilepsy (P < .05). As seen more commonly in patients with diffuse pathologies and intractable frequent seizures, mental retardation was detected by WISC-R in 64.5% of patients (P < .05). CONCLUSION: In cases with cortical developmental malformation, epilepsy/EEG features and neurodevelopmental prognosis can be predicted depending on the developmental process and type and extent of involvement. Patients should be followed up closely with EEG.

Structural changes in the brain on cranial magnetic resonance imaging in severely malnourished children before and after treatment.

OBJECTIVE: The aim of this study was to examine the structural changes in brain on magnetic resonance imaging (MRI) in severely malnourished children before and after treatment. METHODS: This was a prospective study conducted at a Severe Malnutrition Treatment Unit (SMTU) at a tertiary care center. The study was carried out with 52 severely malnourished children 0 to 5 y of age admitted to the SMTU. The cases with neurologic diseases such as epilepsy, cerebral palsy, meningitis, tumor, or hydrocephalus; or history of perinatal asphyxia were excluded to enable us to detect central nervous system changes attributable to malnutrition only. MRI brain without contrast was performed on all 52 cases at admission and after 6 mo of treatment and were reported by an experienced radiologist. The common MRI findings in the study included cerebral atrophy, dilated ventricles, periventricular white matter changes, widened cortical sulci, enlarged basal cisterns, and widened interhemispheric fissure. Development assessment using the Denver Developmental Screening Test II was also performed to infer correlation with MRI findings. RESULTS: Of the 52 children who were severely malnourished, 10 (i.e., 19.2%) had abnormal MRI findings. Development assessment revealed that of the 10 abnormal MRI cases, 80% had delayed developmental milestones as well. These changes are reversible in most cases after nutritional rehabilitation. CONCLUSIONS: The present study revealed that malnutrition per se has a significant effect on the structure of the developing brain and also affects neurologic development. These changes are reversible after treatment so early stimulation and nutritional rehabilitation can help to prevent permanent neurologic derangements.

Emerge of NDM-1-Producing Multidrug-Resistant Pseudomonas aeruginosa and Co-Harboring of Carbapenemase Genes in South of Iran.

BACKGROUND: New Delhi metallo-beta-lactamase-1 (NDM-1) is one of the most important emerging antibiotic resistance. Co-harboring three or four carbapenemases is rare and only a few reports exist in the literature. We described the characteristics of the large epidemic outbreaks and reports co-producing blaNDM-1 with the other carbapenemase genes in P. aeruginosa isolates. METHODS: This present cross-sectional research was conducted on 369 P. aeruginosa isolates obtained from burn and general hospitals within years 2013 to 2016. Beta-lactamase classes A, B and D genes were identified by PCR method. Modified hodge test (MHT), double-disk potentiation tests (DDPT) and double disk synergy test (DDST) were performed for detection carbapenemase and metallo beta-lactamase (MBL) production of blaNDM-1 positive P. aeruginos isolates. RESULTS: From 236 carbapenem-resistant P. aeruginosa (CRPA), 116 isolates have had MBL genes and twenty-nine isolates were found positive for blaNDM-1 . In CRPA isolates, blaIMP-1 , blaVIM-2 and blaOXA-10 were identified in 27.5%, 21.1% and 32.2% of isolates respectively, while co-producing blaNDM-1 , blaIMP-1 , blaOXA-10 , co-producing blaNDM-1 , blaVIM-2 , blaOXA-10 and co-producing blaIMP-1 , blaVIM-2 were determined in 11 (4.6%), 8 (3.4%) and 27 (11.4%) of isolates respectively. CONCLUSION: The finding of this co-existence of multiple carbapenemase resistance genes is threating for public health. Dipicolinic acid is a superior MBL inhibitor in DDPT antique than EDTA in DDST method for the detection of MBL-blaNDM-1 producing P. aeruginosa.

Prevalence of Anemia and Its Associated Risk Factors Among 6-Months-Old Infants in Beijing.

Objective: The worldwide prevalence of anemia is ~24.8%. Iron deficiency anemia is common in children and women and associated with sensory, motor, cognitive, language, and socioemotional deficits. Therefore, detection and early intervention strategies for anemia in infants are urgently needed. To prevent the occurrence of iron deficiency anemia, we aimed to identify risk factors associated with anemia in infants. Methods: This investigation involved a cross-sectional study of 6-months-old infants discharged between April 2014 and September 2017 from Peking University First Hospital. We assessed birth information, maternal age, and maternal educational level as well as data on feeding style, complementary foods and primary caregivers. The infants were assessed with the Denver Developmental Screening Test (DDST). Results: A total of 1,127 6-months-old infants were enrolled at the hospital. We found that the prevalence of anemia among infants in Beijing was ~11.8%. Premature infants had a higher rate of anemia than full-term infants (χ2 = 40.103, P < 0.001). Infants born in autumn or winter were at an elevated risk of developing anemia (χ2 = 22.949, P < 0.001). Birth weight had no effect on the rate of anemia in infants (χ2 = 0.023, P = 0.568). Infants who were exclusively breastfeeding had higher anemia rates than those who were fed formula (χ2 = 38.466, P < 0.001). Infants whose caregivers added no complementary foods had higher anemia rates (24.7%) than those whose caregivers added more than two kinds of complementary food (8.2%). The type of caregiver had no effect on the anemia rate in infants (χ2 = 0.031, P = 1.000). Conclusions: The following factors resulted in a higher prevalence of anemia in our study a gestational age at birth of <37 weeks, exclusive breastfeeding, a lack of supplementation with complementary foods and a spring birth date. No significant differences in DDST pass rates were evident between infants with and without anemia.

Corrigendum: Prevalence of Anemia and Its Associated Risk Factors Among 6-Months-Old Infants in Beijing.

[This corrects the article DOI: 10.3389/fped.2019.00286.].

Clinical characteristics and course of oral somatic delusions: a retrospective chart review of 606 cases in 5 years.

OBJECTIVE: Oral cenesthopathy is characterized by foreign body sensations without medical and dental evidence for them. It is thought to be a rare disease in psychiatry, but many patients are visiting dental clinics seeking treatment to remove a foreign body. Even though the features of oral cenesthopathy might be different between a psychiatric clinic and a dental clinic, there has been no clinic-statistical study from dentists. In this study, we report a clinico-statistical study of patients with oral cenesthopathy in dentistry. METHODS: This is a retrospective chart review of 606 outpatients with oral cenesthopathy in Tokyo Medical and Dental University from April 2010 through to March 2015. RESULTS: A total of 159 male and 447 female patients were included in this study. The mean age was 62.08 years, and female patients were older than male patients. The trigger of the dental treatment and the acute phase of depression at the onset were significantly related (p=0.037). Only 128 patients (36%) had clinically significant improvement after 6 months of pharmacotherapy. No history of psychiatric disorders (odds ratio [OR] 0.479 [95% confidence interval {CI}: 0.262-0.875], p=0.017) and longer duration of illness (>18 months) (OR 2.626 [95% CI: 1.437-4.799], p=0.002) were significant factors for clinical outcomes. CONCLUSION: Patients with oral cenesthopathy in our clinic were predominantly elderly female patients. Dental treatment in the acute phase of depression might be a risk factor for oral cenesthopathy. Therefore, comprehending the situation of psychiatric disorder and obtaining adequate informed consent might be required to prevent the trouble concerning oral cenesthopathy.

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.

Comparision of Three Laboratory Tests for Detection of AmpC ß Lactamases in Klebsiella Species and E. Coli.

BACKGROUND AND OBJECTIVE: AmpC ß lactamases are one of the important causes of drug resistance in gram negative bacteria. Failure to detect these enzymes in the laboratory has contributed to therapeutic failures but there are till date no standard guideline available. This study was therefore undertaken to evaluate three phenotypic laboratory tests and the inhibitors used in two of the tests to detect AmpC ß lactamases produced by E. coli and Klebsiella species as they are most commonly isolated organisms. METHODS: E. coli and Klebsiella isolates from different clinical samples were tested for ESBLs production as per CLSI guidelines and excluded from the study. The non-ESBLs isolates were then screened for AmpC ß lactamases production, by cefoxitin and then confirmed by three different methods, i.e., Disc Potentiation Test (DPT) , Double Disc Synergy Test (DDST) and Modified Three Dimensional Test (M3DT) which in the absence of molecular methods, was taken as the gold standard. Boronic acid and cloxacillin were used as inhibitory agents in the Disc Potentiation and Double Disc synergy Tests. RESULTS: A total of 2,933 isolates were tested out of which 165 isolates were detected as non ESBLs producers,135 (81.82%) when screened for AmpC ß lactamases based on resistance to cefoxitin were labelled as positive. 30 (18.18%) cefoxitin sensitive isolates were labelled as probably non AmpC producers . M3DT, in addition to detecting all the 135 (100%) cefoxitin resistant isolates, also detected 5 (16.67%) cefoxitin sensitive isolates as AmpC producers. Other phenotypic tests, DPT and DDST with different inhibitors like boronic acid and cloxacillin in different potencies were all found to be less sensitive. The best results among these two methods were obtained with DDST using cloxacillin 500µg. CONCLUSION: In the absence of recommended guidelines for AmpC detection, the study reports, among the tests performed, M3DT as the best phenotypic method for AmpC confirmation, as it is not only the most sensitive but also specific test for AmpC as it rules out the resistance due to other mechanisms like the porin channel.

Prevalence and antibiogram of Extended Spectrum ß-Lactamase (ESBL) producing Gram negative bacilli and further molecular characterization of ESBL producing Escherichia coli and Klebsiella spp.

BACKGROUND AND OBJECTIVES: Resistance to a wide variety of common antimicrobials has made the proliferation of Extended spectrum ß-lactmase (ESBL) producing strains a serious global health concern that has complicated treatment strategies. The high proportion of ESBL producers among the Enterobacteriaceae and the complex molecular epidemiology with diverse types of ESBL genes are alarming. This study was undertaken to identify ESBL production in various Gram negative bacilli isolated and to further characterize ESBL producers among Escherichia coli and Klebsiella spp by PCR, which were initially screened by phenotypic method. MATERIALS AND METHODS: A total of 722 isolates of Gram negative bacilli were isolated. Presence of ESBL positivity was detected using the double disk synergy test (DDST). Their antibiogram was studied. PCR analysis for ß-lactamase (bla) genes of the family TEM, SHV and CTX-M was also carried out using designed primers in 20 ESBL isolates each of Escherichia coli and Klebsiella spp. RESULTS: Among 722 Gram negative bacilli isolated 379 (52.49%) were ESBL producers. The major source of ESBL producers were respiratory tract samples, highest ESBL production was observed in Klebsiella sp. (67.04%). Resistance to multiple classes of antibiotics was observed among ESBL producers. Among ESBL producing genes prevalence of bla-CTX-M (82.5%) was highest, followed by bla-TEM (67.5%) and bla-SHV (57.5%) in the present study. The frequency of ESBL producing strains among clinical isolates has been steadily increasing. Advance drug resistance surveillance and molecular characteristics of ESBL isolates is necessary to guide the appropriate and judicious antibiotic use.

The impact of anemia and body mass index (BMI) on neuromotor development of preschool children

Summary Objective: According to data from the World Health Organization (WHO), anemia is a prevalent health problem that leads to increased morbidity and mortality, especially in preschool children. Anemia is recognized as a major health problem due to its negative effects on the mental and physical development during childhood. The aim of our study was to determine the levels of anemia of children in a kindergarten affiliated to the Directorate of National Education using a non-invasive method, and to investigate the effects of anemia on the physical, mental and neuromotor development of children. Method: The levels of anemia was evaluated by using a non-invasive measurement device. Data collection was performed by means of a questionnaire to evaluate the children's physical development and set Denver Developmental Screening Test II scores. Results: Our findings show that 21% of non-anemic and 15% of anemic children are in the suspected abnormal group according to their DDST II total score. Furthermore, it has been identified that mild anemia has a positive effect on neuromotor development, while overweight and obesity affect neuromotor development in a negative way. Conclusion: According to the results obtained from the study, mild anemia may have a positive effect on the children's neuromotor development, while malnutrition could have a negative impact.