Staphylococcus aureus Induced Wound Infections Which Antimicrobial Resistance, Methicillin- and Vancomycin-Resistant: Assessment of Emergence and Cross Sectional Study.

Background: Wound infection is a prevalent concern in the medical field, being is a multi-step process involving several biological processes. Methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) infections often occur in areas of damaged skin, such as abrasions and open wounds. Methods: This research aims to light the incidence of MRSA and VRSA in wound swabs, the antimicrobial susceptibility configuration of isolated S. aureus patterns in pus/wound samples collected from Saudi Arabian tertiary hospital. The cross section study, ß- lactamase detection, VRSA genotyping, MAR index, D-test and VRSA genotyping are methods, which used for completed this research. Results: Patients of several ages and genders delivered specimens from two hospitals in the Al jouf area, in the northern province of Saudi Arabia. S. aureus was found in 188 (34.7%) of the 542 wounds. The traumatized wounds provided 71 isolates (38.8%), surgical wound provided 49 isolates (26.8%) and abscess were represented 16 by isolates (8.7%). In the study, 123 (65.4%) out of 188 were MRSA, 60 (31.9%) were MSSA, and five (2.7%) were VRSA. Linezolid and rifampin were found to be the most effective antimicrobials with 100% in vitro antibacterial activity against S. aureus isolates. The Multiple antimicrobials resistance (MAR) index revealed 73 isolates (38.9%) with a MAR index greater than 0.2, and 115 (61.1%) less than 0.2. The D-test showed that of MLSb phenotypes among S. aureus, 22 (11.7%) strains were D-test positive (MLSbi phenotype), 53 (28.2%) strains were constitutive MLSc phenotypes, and 17 (9%) strains were shown to have MSb phenotypes. All VRSA isolates (n=5) were found to be positive for vanA, and no vanB positive isolates were detected in the study. Conclusion: Regular monitoring and an antimicrobials stewardship program should be in place to provide critical information that can be utilized for empirical therapy and future prevention strategies.

Developmental Eye Movement (DEM) and King-Devick (K-D) Performance in Multiple Sclerosis.

Eye movement disorders have been reported in patients with multiple sclerosis (MS) as saccadic disturbances. Several methods have been described for the assessment of saccades, including the K-D and DEM tests. The performance of these tests also involves attention, language, and other brain areas which have been reported to be altered in MS patients. The aim of the study was to determine how developmental eye movement (DEM) and King-Devick (K-D) tests are affected in MS-patients. It was also to analyze whether a resolved episode of optic neuritis (ON) has a negative influence. Subjects with MS showed worse outcomes (higher times) in DEM test (72 (26) s and a K-D test (56 (22) s compared to healthy subjects (64 (7) s and 55 (11) s, respectively). However, a previous ON episode did not worsen the MS-times of DEM (80 (33) s or of K-D (62 (33) s. Horizontal saccades with the DEM showed differences between subjects with MS (with and without ON) and healthy ones (p < 0.05), whereas no such differences were found in the vertical saccades. According to the DEM instructions, MS patients would present heterogeneous oculomotor and non-visual disturbances. Regarding the K-D test, only the third card (the most complex one) showed differences (p < 0.05) between groups. These tests can capture impairment of attention, language, and other areas that correlate with suboptimal brain function in addition to the oculomotor dysfunctions present in subjects with MS.

Huntsman spider phylogeny informs evolution of life history, egg sacs, and morphology.

Huntsman spiders (Araneae: Sparassidae) are among the most speciose spider families, with a near-worldwide distribution, diverse habitats, equally diverse life histories, and five prolonged subsocial species. Previous molecular phylogenies have focused on individual subfamilies or clades. Here, we provide a phylogenetic inference with broadened sampling from 37 genera and eight of the eleven sparassid subfamilies. We increased taxon sampling by including species not previously sequenced and most available data on GenBank of two mitochondrial (COI, 16S rRNA) and two nuclear (H3, 28S rRNA) genes for a total of 262 ingroup taxa and nine outgroup taxa. Divergence dates were estimated using outgroup fossil taxa suggesting the sparassids evolved âˆ¼ 100 mya (stem age), while the clade containing all subfamilies except Sparianthinae evolved âˆ¼ 90 mya (stem age). Using a stochastic map approach with 40 species, this is the first sparassid phylogeny to incorporate extensive biology and life history data. Correlations of life history traits with solitary, subsocial, and prolonged subsocial behavior are examined using the D-test. Sparassid sociality is associated with life history traits that allow developing spiders to remain in their natal retreat longer (e.g., larger permanent retreats, plastered egg sacs, and ontogenetically delayed foraging), but is unrelated to body size or lifespan. Detailed morphological scoring of the endemic Australian subfamily Deleninae contextualizes existing molecular data, including in the Isopeda-Holconia-Isopedella complex. This study supports the monophyly of many major lineages, including for the first time, the Sparianthinae, but indicates multiple clades (Sparassinae and Eusparassinae) are paraphyletic and need further revision.

Comparison of Phenotypic and Genotypic Characterization Methods for the Detection of Methicillin-Resistant Staphylococcus Aureus.

Introduction Methicillin-resistant Staphylococcus aureus (MRSA) is associated with high morbidity and mortality due to the development of antimicrobial resistance secondary to irrational use of antibiotics, nonadherence to infection control practices, and increased use of intravascular devices in healthcare systems. Detection of MRSA is critical in clinical microbiology laboratories as it helps identify MRSA carriers and avoid treatment failure in patients. Hence, this study compared various phenotypic methods with the standard genotyping method to determine a method that permits rapid and accurate detection of MRSA. Materials & Methods Staphylococcus aureus (S. aureus) was initially identified based on colony morphology, Gram staining, standard biochemical tests, and antibiotic susceptibility using disk diffusion. MRSA was identified based on the detection of the mecA gene by polymerase chain reaction (PCR) and subsequent gel electrophoresis. Disk diffusion using cefoxitin or oxacillin and mannitol salt agar with 6-µg/ml oxacillin were used for phenotypic detection of MRSA. The D test was used to detect inducible clindamycin resistance in S. aureus isolates. Results Of the 100 S. aureus isolates analyzed, 37% were identified as MRSA by PCR and the cefoxitin disk diffusion method; however, only 31% were detected by the oxacillin disk diffusion method and 29% by the mannitol salt agar method. The sensitivity of the cefoxitin disk diffusion test, oxacillin disk diffusion, and mannitol salt agar methods was 86.05%, 83.78%, and 70.73%, respectively. Specificity was 100% for all the three phenotypic methods (p < 0.001). Notably, inducible clindamycin resistance was found in 37.2% of the MRSA isolates, indicating potential challenges in treatment. Conclusion Among the three phenotypic methods tested, the cefoxitin disk diffusion method had 100% sensitivity and specificity, which is similar to that of PCR-based MRSA detection. Hence, the cefoxitin disk diffusion method is recommended for use in clinical laboratories, where molecular methods are not available as it is both cost-effective and easy to perform.

Huntsman spider phylogeny informs evolution of life history, egg sacs, and morphology

Huntsman spiders (Araneae: Sparassidae) are among the most speciose spider families, with a near-worldwide distribution, diverse habitats, equally diverse life histories, and five prolonged subsocial species. Previous molecular phylogenies have focused on individual subfamilies or clades. Here, we provide a phylogenetic inference with broadened sampling from 37 genera and eight of the eleven sparassid subfamilies. We increased taxon sampling by including species not previously sequenced and most available data on GenBank of two mitochondrial (COI, 16S rRNA) and two nuclear (H3, 28S rRNA) genes for a total of 262 ingroup taxa and nine outgroup taxa. Divergence dates were estimated using outgroup fossil taxa suggesting the sparassids evolved ∼100 mya (stem age), while the clade containing all subfamilies except Sparianthinae evolved ∼90 mya (stem age). Using a stochastic map approach with 40 species, this is the first sparassid phylogeny to incorporate extensive biology and life history data. Correlations of life history traits with solitary, subsocial, and prolonged subsocial behavior are examined using the D-test. Sparassid sociality is associated with life history traits that allow developing spiders to remain in their natal retreat longer (e.g., larger permanent retreats, plastered egg sacs, and ontogenetically delayed foraging), but is unrelated to body size or lifespan. Detailed morphological scoring of the endemic Australian subfamily Deleninae contextualizes existing molecular data, including in the Isopeda-Holconia-Isopedella complex. This study supports the monophyly of many major lineages, including for the first time, the Sparianthinae, but indicates multiple clades (Sparassinae and Eusparassinae) are paraphyletic and need further revision.

Distribution of Macrolide-Lincosamide-Streptogramin B Antibiotics Resistance Genes in Clinical Isolates of Staphylococci.

BACKGROUND: Staphylococci are posing threat due to increasing trend of antimicrobial resistance particularly methicillin. Macrolide lincosamide streptogramin B (MLSB) family of antibiotics is commonly used to treat such infections. This study was aimed to determine the prevalence of inducible clindamycin resistance and observation of erm and msr genes among Staphylococci isolated from tertiary care hospital of Nepal during July 2017 to March 2018. METHODS: Staphylococci from different clinical specimens were identified and antibiotic susceptibility profile was assessed following Kirby Bauer disc diffusion method. The double disc diffusion or D-zone test as outlined in CLSI document M100-S24 was performed to examine inducible clindamycin resistant isolates. Multiplex PCR was performed for detection of erm and msr gene in isolates using specific primers for ermA, ermB, ermC, msrA and msrB genes. RESULTS: Of the 60 Staphylococci isolates, 39 (65%) were S. aureus and 21 (35%) were coagulase negative Staphylococci (CNS) with 25 (64%) and 15 (71%) representing methicillin resistant S. aureus and CNS respectively. Constitutive and inducible MLSB phenotype was observed among 24 (40%) and 14 (23%) isolates respectively by D test. The most prevalent resistant gene was ermC (37%) followed by msrB (12%), ermB (10%) and msrA (10%). None of the isolates were found to possess ermA gene. CONCLUSIONS: The presence of constitutive and inducible MLSB as well as resistant genes among Staphylococci necessitates detection of such isolates to minimize treatment failure. The result from this study may help elucidate the predominant resistant characteristics in clinical Staphylococci isolated from tertiary care hospital of Nepal.

Macrolide-lincosamide-streptogramin B resistance phenotypes and their associated genotypes in Staphylococcus aureus isolates from a tertiary level public hospital of Uruguay / Fenotipos de resistencia a macrólidos lincosamidas-estreptograminas B y sus genotipos asociados en Staphylococcus aureus aislados en un hospital público de nivel terciario en Uruguay

Abstract This study was undertaken to investígate the resistance phenotypes to macrolide-lincosamide-streptogramin B (MLSb) antibiotics and their associated genotypes in isolates of Staphylococcus aureus. We analyzed one hundred, consecutive, non-duplicate isolates (methicillin-susceptible MSSA, n = 53 and methicillin-resistant MRSA, n =47) obtained from var-ious clinical samples between July 2012 to December 2013. The resistance profile to MLSb antibiotics was determined by phenotypic methods and the resistance genes were detected by PCR assays. All of the isolates were subjected to pulsed-field gel electrophoresis (SmaI-PFGE). The overall prevalence of resistance to MLSb antibiotics was 38% and the resistance phenotype distribution was as follows: cMLSb, 22%; iMLSB, 10%; MSb, 5% and L, 1%. We detected ermA, ermC, ermB and mrsA/B genes in these resistant isolates. The single ermA gene was commonly observed mainly in those with a cMLSb R phenotype, whereas the combination ermA and ermC was more commonly observed in isolates with inducible expression. The patterns of SmaI-PFGE suggest a great genetic diversity in both MRSA and MSSA resistant to MLSb antibiotics. The results demonstrate the local presence of S. aureus resistant to MLSb antibiotics and its most frequently described responsible genes. Some of these isolates, especially those with the iMLSB phenotype, may be associated with therapeutic failure. Therefore, efforts should be directed to the correct detection of all MLSb resistant isolates using appropriate laboratory tests. PFGE results reveal a wide spread of resistance genes rather than the circulation of S. aureus clones resistant to MLSb antibiotics.

Resumen Los objetivos de este estudio fueron investigar en Staphylococcus aureus la presencia de fenotipos resistentes a los antibióticos macrólidos, lincosamidas y estreptograminas tipoB (MLSb) y conocer sus genotipos responsables. Analizamos 100 aislamientos consecutivos, no duplicados (53 sensibles a meticilina [MSSA] y 47 resistentes a meticilina [MRSA]), obtenidos entre 2012 y 2013 a partir de diferentes muestras clínicas. El perfil de resistencia a los antibióticos MLSb fue determinado por métodos fenotípicos y los genes de resistencia se detectaron por PCR. Todos los aislamientos fueron comparados por SmaI-PFGE. La prevalencia global de resistencia a los antibióticos MLSB fue del 38% y la distribución de los fenotipos de resistencia fue la siguiente: cMLSB, 22%; iMLSB, 10%; MSB, 5%; L, 1%. Se detectaron los genes ermA, ermC y mrsA/B en los aislamientos resistentes. El gen ermA se observó, sobre todo, en aislamientos con fenotipo resistente constitutivo R (cMLSB), mientras que la combinación ermA y ermC se detectó principalmente en aislamientos con resistencia inducible (iMLSB). Los patrones de Smal-PFGE sugieren una gran diversidad genética en los aislamientos resistentes a los antibióticos MLSb, tanto MRSA como MSSA. Los resultados demuestran la presencia local de S. aureus resistentes a los antibióticos MLSB y de sus genes responsables más frecuentemente descritos. Estos cultivos, especialmente aquellos con fenotipo resistente iMLSB, pueden asociarse con fallas terapéuticas. Por lo tanto, los esfuerzos deben dirigirse a la correcta detección de todos los cultivos resistentes a MLSB utilizando pruebas de laboratorio adecuadas. Los resultados de Smal-PFGE sugieren una amplia diseminación de genes de resistencia, más que la circulación de clones resistentes a los antibióticos MLSB.

Domestic wastewater treatment plants as sources of macrolide-lincosamide-streptogramin B- and penicillin-resistant Staphylococcus aureus in Brazil / Plantas de tratamiento de aguas residuales domésticas como fuentes de Staphylococcus aureus resistente a macrólidos-lincosamida-estreptogramina B- y penicilina en Brasil / Estações de tratamento de águas residuais domésticas como fontes de Staphylococcus aureus resistente a macrolídeo-lincosamida-estreptograma e B- e penicilina no Brasil

SUMMARY Staphylococcus aureus is one of the main bacteria that affect human health. Its reduced susceptibility to beta-lactam antibiotics has driven the clinical use of macrolides and lincosamides. However, the presence of macrolide-lincosamide-streptogramin B (MLSB)-resistant S. aureus strains is increasingly common. Wastewater treatment plants (WWTPs) are the main anthropogenic source of resistance determinants. However, few studies have assessed the importance of this environment on the dissemination of MLSB-resistant S. aureus strains. Thus, we aimed to evaluate the impact of a domestic WWTP on the resistance to MLSB and penicillin in S. aureus in southeast Brazil. Of the 35 isolates tested, 40.6% were resistant to penicillin. Resistance to erythromycin (8.6%) and quinolones (2.8%) was less common. Despite the low rate of resistance to clindamycin (2.8%), many isolates showed reduced susceptibility to this antibiotic (57.1%). Regarding the resistance phenotypes of staphylococci isolates, inducible MLSB resistance (D-test positive) was found in two isolates. In addition, 27 S. aureus isolates showed the ability to produce penicillinase. In this article, we report for the first time the importance of WWTPs in the dissemination of MSLB resistance among S. aureus from southeast Brazil.

RESUMEN Staphylococcus aureus es una de las principales bacterias que afectan la salud humana. Su susceptibilidad reducida a los antibióticos betalactámicos ha impulsado el uso clínico de macrólidos y lincosamidas. Sin embargo, la presencia de cepas resistentes a macrólido-lincosamida-estreptogramina B (MLSB) de S. aureus es cada vez más común. Las plantas de tratamiento de aguas residuales (PTAR) son la principal fuente antropogénica de determinantes de resistencia. Sin embargo, pocos estudios han evaluado la importancia de este entorno en la diseminación de cepas de S. aureus resistentes a MLSB. Nuestro objetivo fue evaluar el impacto de una PTAR doméstica en MLSB y la resistencia a la penicilina en S. aureus en el sureste de Brasil. De los 35 aislamientos analizados, el 40,6% eran resistentes a la penicilina. La resistencia a la eritromicina (8,6%) y quinolonas (2,8%) fue menos común. A pesar de la baja tasa de resistencia a la clindamicina (2,8%), muchos aislamientos mostraron sensibilidad reducida a este antibiótico (57,1%). Con respecto a los fenotipos de resistencia de los aislamientos de estafilococos, la resistencia inducible a MLSB (prueba D positiva) se encontró en dos aislamientos. Además, 27 aislamientos de S. aureus mostraron la capacidad de producir penicilinasa. En este artículo informamos, por primera vez, la importancia de las PTAR en la difusión de la resistencia a MSLB entre S. aureus del sureste de Brasil.

RESUMO O Staphylococcus aureus é uma das principais bactérias que afetam a saúde humana. Sua reduzida suscetibilidade aos antibióticos beta-lactâmicos tem impulsionado o uso clínico de macrolídeos e lincosamidas. No entanto, a presença de cepas de S. aureus resistentes a macrolídeo-lincosamida-estreptogramina B (MLSB) é cada vez mais comum. As estações de tratamento de esgoto (ETEs) são a principal fonte antropogênica de determinantes de resistência. No entanto, poucos estudos avaliaram a importância desse ambiente na disseminação de cepas de S. aureus resistentes ao MLSB. Assim, nosso objetivo foi avaliar o impacto de uma ETE doméstico na resistência ao MLSB e à penicilina em S. aureus no sudeste do Brasil. Dos 35 isolados testados, 40,6% eram resistentes à penicilina. Resistência à eritromicina (8,6%) e quinolonas (2,8%) foi menos comum. Apesar da baixa taxa de resistência à clindamicina (2,8%), muitos isolados apresentaram sensibilidade reduzida a esse antibiótico (57,1%). Em relação aos fenótipos de resistência dos isolados de estafilococos, a resistência induzível ao MLSB (D-teste positivo) foi encontrada em dois isolados. Além disso, 27 isolados de S. aureus mostraram a capacidade de produzir penicilinase. Neste artigo relatamos pela primeira vez a importância das ETEs na disseminação da resistência do MSLB entre S. aureus do sudeste do Brasil.

Phenotypic and Genotypic Characterization of Resistance to Macrolides and Lincosamides in Streptococcus agalactiae Isolated from Pregnant Women in Misiones, Argentina.

The aim of this work was to determine the susceptibility, molecular profile, and clonal relationship in Streptococcus agalactiae (group B Streptococcus [GBS]) isolated from vaginal-rectal swab samples. We worked with 200 isolates collected from pregnant women between 35 and 37 weeks of gestation. The macrolide-lincosamide-streptogramin B (MLSB) resistance phenotypes were determined using the double-disc assay. Susceptibility to erythromycin (ERI) and clindamycin (CLI) was performed with the E-test. Resistance genes ermB and ermTR were detected by polymerase chain reaction. Clonal studies were performed using the random amplification of polymorphic DNA. Twelve (6%) of the isolates were resistant to ERI and 10 (5%) of them to CLI. Fifty percent of the resistant strains corresponded to serotype III, 25% to serotype V, and the remaining 25% to serotype Ia, II, and nontypeable strains. The cMLSB phenotype was detected in eight strains (66.67%) and the iMLSB phenotype in four (33.33%). The minimum inhibitory concentration values were between 1.5 and 16 µg/mL for ERI, and between 1 and 32 µg/mL for CLI. Out of the 25 strains susceptible to ERI and CLI, the presence of the ermB gene was detected in eight of them and the ermTR gene in one strain. The ermB gene was detected in the 12 strains that initially had some macrolide resistance phenotype. The ermTR gene was detected in three out of the four strains with the iMLSB phenotype. The resistance to macrolides in the province of Misiones is due to multiclonal spread. The phenotypic and genotypic characterization of macrolide resistance in GBS strains are crucial to contribute to the correct intrapartum prophylactic antibiotic therapy of allergic pregnant women and the epidemiological surveillance of these strains.

Assessment of inducible clindamycin resistance and Hyper Variable Region (HVR) of mecA gene in clinical staphylococci.

OBJECTIVE: To study the prevalence of inducible clindamycin along with vancomycin and methicillin resistance and assessment of hyper variable region (HVR) of mecA gene among different clinical isolates of Staphylococcus spp. METHODS: A total of 176 clinical isolates of Staphylococci were collected from Pakistan Institute of Medical Sciences (PIMS), Islamabad during 2014-2015. The sample sources were pus, blood, urine, sputum, tracheal secretions and tissue fluids. Bacterial identification was done by colony morphology and biochemical tests. Kirby-Bauer disc-diffusion method was carried out to assess the susceptibility against different antibiotics. Minimal inhibitory concentrations (MICs) were done for vancomycin resistance. Double Disk Diffusion test (D-test) was used to detect the clindamycin inducible resistance. PCR was performed to detect erm(C), mecA and HVR genes. RESULTS: Clindamycin inducible resistance among Staphylococcal isolates was found to be 7%, whereas in S. aureus it was 4%, and in coagulase negative Staphylococci (CoNS) it was 11%. The highest resistance was observed against fosfomycin, fusidic acid and cefoxitin. Vancomycin resistance was observed in 23 isolates (13%) of Staphylococci. erm(C), mecA and HVR genes were found in 18%, 50% and 42% respectively. CONCLUSIONS: D-test must be performed routinely to avoid clindamycin failure. A high level of resistance against vancomycin in Staphylococcal isolates is a concern for public health.

Macrolide-lincosamide-streptogramin B resistance phenotypes and their associated genotypes in Staphylococcus aureus isolates from a tertiary level public hospital of Uruguay.

This study was undertaken to investigate the resistance phenotypes to macrolide-lincosamide-streptogramin B (MLSB) antibiotics and their associated genotypes in isolates of Staphylococcus aureus. We analyzed one hundred, consecutive, non-duplicate isolates (methicillin-susceptible MSSA, n=53 and methicillin-resistant MRSA, n=47) obtained from various clinical samples between July 2012 to December 2013. The resistance profile to MLSB antibiotics was determined by phenotypic methods and the resistance genes were detected by PCR assays. All of the isolates were subjected to pulsed-field gel electrophoresis (SmaI-PFGE). The overall prevalence of resistance to MLSB antibiotics was 38% and the resistance phenotype distribution was as follows: cMLSB, 22%; iMLSB, 10%; MSB, 5% and L, 1%. We detected ermA, ermC, ermB and mrsA/B genes in these resistant isolates. The single ermA gene was commonly observed mainly in those with a cMLSB R phenotype, whereas the combination ermA and ermC was more commonly observed in isolates with inducible expression. The patterns of SmaI-PFGE suggest a great genetic diversity in both MRSA and MSSA resistant to MLSB antibiotics. The results demonstrate the local presence of S. aureus resistant to MLSB antibiotics and its most frequently described responsible genes. Some of these isolates, especially those with the iMLSB phenotype, may be associated with therapeutic failure. Therefore, efforts should be directed to the correct detection of all MLSB resistant isolates using appropriate laboratory tests. PFGE results reveal a wide spread of resistance genes rather than the circulation of S. aureus clones resistant to MLSB antibiotics.

Inducible clindamycin resistance and erm genes in Staphylococcus aureus in school children in Kathmandu, Nepal.

AIM: Resistance to methicillin and Macrolide-Lincosamide and Streptogramins B and their association with erm genes in Staphylococcus aureus are unknown in Nepal. MATERIALS & METHODS: Nonduplicate nasal swabs from 160 school children were collected from April to September 2018 and processed using standard microbiological procedures. RESULTS: Out of 160 samples, 64 (40%) were S. aureus in which 17 (26.6%) were methicillin-resistance Staphylococcus aureus (MRSA). D-test identified 15 (23.4%) as inducible clindamycin-resistant, which were more prevalent in MRSA (76.4%) than methicillin-sensitive S. aureus (MSSA; 4.2%). 18.7% of isolates harbored the ermC gene followed by ermA (15.6%) and ermB (3.1%), and were more in MRSA than MSSA. CONCLUSION: To prevent treatment failure by inducible resistance, D-test must be performed on erythromycin-resistant and/or clindamycin-sensitive isolates.

Nanoporous gold: a hierarchical and multiscale 3D test pattern for characterizing X-ray nano-tomography systems.

Full-field transmission X-ray microscopy (TXM) is a well established technique, available at various synchrotron beamlines around the world as well as by laboratory benchtop devices. One of the major TXM challenges, due to its nanometre-scale resolution, is the overall instrument stability during the acquisition of the series of tomographic projections. The ability to correct for vertical and horizontal distortions of each projection image during acquisition is necessary in order to achieve the effective 3D spatial resolution. The effectiveness of such an image alignment is also heavily influenced by the absorption properties and strong contrast of specific features in the scanned sample. Here it is shown that nanoporous gold (NPG) can be used as an ideal 3D test pattern for evaluating and optimizing the performance of a TXM instrument for hard X-rays at a synchrotron beamline. Unique features of NPG, such as hierarchical structures at multiple length scales and high absorbing capabilities, makes it an ideal choice for characterization, which involves a combination of a rapid-alignment algorithm applied on the acquired projections followed by the extraction of a set of both 2D- and 3D-descriptive image parameters. This protocol can be used for comparing the efficiency of TXM instruments at different synchrotron beamlines in the world or benchtop devices, based on a reference library of scanned NPG samples, containing information about the estimated horizontal and vertical alignment values, 2D qualitative parameters and quantitative 3D parameters. The possibility to tailor the ligament sizes of NPG to match the achievable resolution in combination with the high electron density of gold makes NPG an ideal 3D test pattern for evaluating the status and performance of a given synchrotron-based or benchtop-based TXM setup.

Elevated markers of brain injury as a result of clinically asymptomatic high-acceleration head impacts in high-school football athletes.

OBJECTIVEThis prospective observational cohort study of high-school football athletes was performed to determine if high-acceleration head impacts (HHIs) that do not result in clinically diagnosed concussion still lead to increases in serum levels of biomarkers indicating traumatic brain injury (TBI) in asymptomatic athletes and to determine the longitudinal profile of these biomarkers over the course of the football season.METHODSSixteen varsity high-school football athletes underwent baseline neurocognitive testing and blood sampling for the biomarkers tau, ubiquitin C-terminal hydrolase L1 (UCH-L1), neurofilament light protein (NF-L), glial fibrillary acidic protein (GFAP), and spectrin breakdown products (SBDPs). All athletes wore helmet-based accelerometers to measure and record head impact data during all practices and games. At various time points during the season, 6 of these athletes met the criteria for HHI (linear acceleration > 95g and rotational acceleration > 3760 rad/sec2); in these athletes a second blood sample was drawn at the end of the athletic event during which the HHI occurred. Five athletes who did not meet the criteria for HHI underwent repeat blood sampling following the final game of the season. In a separate analysis, all athletes who did not receive a diagnosis of concussion during the season (n = 12) underwent repeat neurocognitive testing and blood sampling after the end of the season.RESULTSTotal tau levels increased 492.6% ± 109.8% from baseline to postsession values in athletes who received an HHI, compared with 164% ± 35% in athletes who did not receive an HHI (p = 0.03). Similarly, UCH-L1 levels increased 738.2% ± 163.3% in athletes following an HHI, compared with 237.7% ± 71.9% in athletes in whom there was no HHI (p = 0.03). At the end of the season, researchers found that tau levels had increased 0.6 ± 0.2 pg/ml (p = 0.003) and UCH-L1 levels had increased 144.3 ± 56 pg/ml (p = 0.002). No significant elevations in serum NF-L, GFAP, or SBDPs were seen between baseline and end-of-athletic event or end-of-season sampling (for all, p > 0.05).CONCLUSIONSIn this pilot study on asymptomatic football athletes, an HHI was associated with increased markers of neuronal (UCH-L1) and axonal (tau) injury when compared with values in control athletes. These same markers were also increased in nonconcussed athletes following the football season.

Phenotypic and genotypic characterization of macrolide resistance among Staphylococcus aureus isolates in Isfahan, Iran.

BACKGROUND AND OBJECTIVES: Macrolide, lincosamide and streptogramin B (MLS B) are noteworthy antibiotics for the treatment of Staphylococcus aureus infections. The purpose of this study, was to determine the phenotypic and genotypic characterization of macrolide resistance, among S. aureus, isolated from clinical samples and nasal swabs. MATERIALS AND METHODS: Totally, 162 non-duplicate S. aureus isolates were collected from clinical samples and nasal swabs, from patients and healthcare workers (HCWs), between March 2016 and September 2016, at four teaching hospitals in Isfahan. The antibiotic resistance profile was determined using disk diffusion test and the presence of resistance genes was detected, using PCR. RESULTS: Of 162 S. aureus isolates, 43.8% (71/162) and 34% (55/162) isolates were erythromycin-resistant and methicillin-resistant S. aureus (MRSA), respectively. The prevalence of constitutive MLS B (cMLS B), inducible MLS B (iMLS B), macrolide-streptogramin B-resistant (MS B) and lincosamide-streptogramin-A resistance (LS A) phenotype was 32%, 6%, 6% and 2%, respectively. The most common erythromycin resistance genes, in S. aureus isolates were ermC (35.2%), followed by ermA (20.4%) and msrA (17.3%). Meanwhile, msrA was detected in 43.6% of MRSA isolates. The frequency of coexistence of ermA+ermC+msrA, in S. aureus isolates was 7% and it was only detected in MRSA isolates. CONCLUSION: In the current study, cMLS B phenotype was the most common erythromycin resistance pattern and ermC was the most prevalent gene in erythromycin-resistant isolates. The results revealed that the various mechanisms of erythromycin resistance are expanding in Isfahan.

A Mainly Circum-Mediterranean Origin for West Eurasian and North African mtDNAs in Puerto Rico with Strong Contributions from the Canary Islands and West Africa.

Maternal lineages of West Eurasian and North African origin account for 11.5% of total mitochondrial ancestry in Puerto Rico. Historical sources suggest that this ancestry arrived mostly from European migrations that took place during the four centuries of the Spanish colonization of Puerto Rico. This study analyzed 101 mitochondrial control region sequences and diagnostic coding region variants from a sample set randomly and systematically selected using a census-based sampling frame to be representative of the Puerto Rican population, with the goal of defining West Eurasian-North African maternal clades and estimating their possible geographical origin. Median-joining haplotype networks were constructed using hypervariable regions 1 and 2 sequences from various reference populations in search of shared haplotypes. A posterior probability analysis was performed to estimate the percentage of possible origins across wide geographic regions for the entire sample set and for the most common haplogroups on the island. Principal component analyses were conducted to place the Puerto Rican mtDNA set within the variation present among all reference populations. Our study shows that up to 38% of West Eurasian and North African mitochondrial ancestry in Puerto Rico most likely migrated from the Canary Islands. However, most of those haplotypes had previously migrated to the Canary Islands from elsewhere, and there are substantial contributions from various populations across the circum-Mediterranean region and from West African populations related to the modern Wolof and Serer peoples from Senegal and the nomad Fulani who extend up to Cameroon. In conclusion, the West Eurasian mitochondrial ancestry in Puerto Ricans is geographically diverse. However, haplotype diversity seems to be low, and frequencies have been shaped by population bottlenecks, migration waves, and random genetic drift. Consequently, approximately 47% of mtDNAs of West Eurasian and North African ancestry in Puerto Rico probably arrived early in its colonial history.

Analysis on effect of blood anti-D antibody detection in infusing RhD positive homotype RBC for patients with RhD negative / 国际检验医学杂志

Objective To investigate the anti-D antibody level after infusing homotype RhD positive RBC in the patient with RhD negative.Methods The clinical data in 20 cases of RhD negative infusing homotype RhD positive RBC in our hospital from January 2010 to January 2016 were collected.The anti-D antibody levels before blood infusion and on 10,20,30,90 d after blood infusion and the titers in the patients with RhD positive were analyzed.Results Among 20 cases of RhD negative infusing RhD positive homotype RBC blood,6 cases were RhD positive within 90 d after blood infusion,in which the RhD positive rates were 25%(3/12) in male and 37.5%(3/8) in female.The anti-D antibody titers in 3 cases of RhD positive were 35,278 and 508 respectively.Conclusion Infusing RhD positive RBC blood in the patients with RhD negative can stimulate the immune mechanism,generates the anti-D antibody at RBC surface.Moreover the RBC phenotype in partial patients with RhD negative may change.

Frequency of genes encoding erythromycin ribosomal methylases among Staphylococcus aureus clinical isolates with different D-phenotypes in Tehran, Iran.

BACKGROUND AND OBJECTIVES: Macrolide, lincosamide and streptogramin type B (MLSB) antibiotics are important in the treatment of Staphylococcus aureus infections and existence of isolates with ability to resist against MLSB antibiotics is worrisome. MATERIALS AND METHODS: In this cross sectional study, 101 S. aureus isolates were collected from patients of five selected hospitals in Tehran over a period of five months. Disk diffusion tests and differentiation between constitutive and inducible resistances were carried out by D-test. The presence of mecA, msrA, ermA and ermC genes were detected using PCR or multiplex PCR. RESULTS: Out of 101 S. aureus isolates, 58 (57.4%) were methicillin resistant and 57 (56.4%) expressed resistance to erythromycin. The prevalence of constitutive MLSB (cMLSB), inducible MLSB (iMLSB) and MS (Negative) phenotype in all erythromycin resistant isolates were 71.9, 26.3 and 1.7%, respectively. Out of all the erythromycin resistant isolates, 57.8% harbored both ermA and ermC genes which possessed constitutive resistance. 8.7% of the isolates contained ermA gene alone which possessed inducible resistance with D phenotype and 5.2% of isolates just contained ermC gene which had inducible resistance with D+ phenotype. msrA gene was detected in 3.5% of the erythromycin resistant S. aureus isolates with constitutive resistance. None of the genes were detected among MS phenotypes. CONCLUSION: In this study, most of S. aureus isolates carried both ermA and ermC genes and there was a significant relationship (P value ≤ 0.05) between different resistance phenotypes and erm genes.

Detection of inducible and constitutive clindamycin resistance among Staphylococcus aureus isolates in a tertiary care hospital, Eastern India.

INTRODUCTION: Clindamycin is an excellent drug for skin and soft tissue Staphylococcus aureus infections, but resistance mediated by inducible macrolide-lincosamide-streptogramin B (iMLSB) phenotype leads to in vivo therapeutic failure even though they may be in vitro susceptible in Kirby-Bauer disk diffusion method. OBJECTIVE: The study was aimed to detect the prevalence of iMLSB phenotype among S. aureus isolates by double disk approximation test (D-test) in a tertiary care hospital, Eastern India. MATERIALS AND METHODS: A total of 209 consecutive S. aureus isolates were identified by conventional methods and subjected to antimicrobial susceptibility testing by Kirby-Bauer disk diffusion method. Erythromycin-resistant isolates were tested for D-test. RESULTS: From 1282 clinical specimens, 209 nonrepeated S. aureus isolates were obtained. Majority of isolates 129 (61.7%) were methicillin-resistant S. aureus (MRSA). There was statistically significant difference between outpatients 60.1% and inpatients 39.9% (P < 0.0001). From 209 S. aureus isolates, 46 (22%) were D-test positive (iMLSB phenotype), 41 (19.6%) were D-test negative (methicillin sensitive [MS] phenotype), and 37 (17.7%) were constitutively resistant (constitutive macrolide-lincosamide-streptogramin B phenotype). The incidence of inducible, constitutive, and MS phenotype was higher in MRSA isolates compared to MS S. aureus (MSSA). The constitutive clindamycin resistance difference between MSSA and MRSA isolates were found to be statistically significant (P = 0.0086). CONCLUSION: The study revealed 22% of S. aureus isolates were inducible clindamycin resistant, which could be easily misidentified as clindamycin susceptible in Kirby-Bauer disk diffusion method. Therefore, clinical microbiology laboratory should routinely perform D-test in all clinically isolated S. aureus to guide clinicians for the appropriate use of clindamycin.

Distribution of erm genes among Staphylococcus aureus isolates with inducible resistance to clindamycin in Isfahan, Iran.

BACKGROUND: The rising frequency of methicillin resistant Staphylococcus aureus (MRSA) has led to an increased use of antibiotics such as macrolide, lincosamide, streptogramin B (MLSB) for the treatment of S. aureus infections. Resistance to MLSB in S. aureus is commonly encoded by erm genes, which can be constitutive MLSB (cMLSB) or inducible MLSB (iMLSB). The purpose of this study was to determine the frequency of cMLSB, iMLSB, and MS phenotypes using D-test and polymerase chain reaction (PCR) methods. MATERIALS AND METHODS: A total of 215 isolates of S. aureus were collected from January 2010 to May 2012 from Al-Zahra Hospital in Isfahan. PCR was performed for detection of mecA gene on all isolates using specific primers. The frequency of MLSB-resistant isolates was determined using D-test, and then a multiplex PCR was performed for detection of ermA, ermB, and ermC genes. RESULTS: Among 215 S. aureus isolates examined, 82 (40.9%) were MRSA, and iMLSB, cMLSB, and MS resistance phenotypes had a frequency of 9 (4.18%), 58 (26.9%), and 11 (5.1%), respectively. Among nine isolates with iMLSB resistance phenotype, four isolates contained ermC gene, two isolates ermB gene, and one isolate ermA gene. Two isolates did not have any erm gene. CONCLUSION: In the current study, cMLSB was the most frequent phenotype and ermC was the most common gene in iMLSB resistant phenotypes.