Morphological and molecular characterization of fungus isolated from tropical bed bugs in Northern Peninsular Malaysia, Cimex hemipterus (Hemiptera: Cimicidae)

Objective: To investigate some morphological and molecular characteristics of fungal parasites isolated from wild tropical bed bug, Cimex hemipterus. Methods: A series of culture methods were used to obtain fungal isolates from field-collected bed bugs. Characteristics of the isolates such as colony appearance, mycelial texture and pigmen-tation were studied to explore their morphology. Isolates were also subjected to a PCRbased genotyping test. Results: There were noticeable differences in morphological characteristics among the four isolates. Conidial areas of one isolate were dark green, whereas those of the remaining colonies were olive-green, black or dark brown. Conidia of the dark green isolate were globose, while those of olive-green, black and dark brown isolates were globose to subglobose, globose to spherical and globose to subglobose/finely roughened, respectively. These morphological specificities and the molecular analyses showed that the fungal internal transcribed spacer ribosomal region and β-tubulin gene sequences of the isolates shared clade with Trichoderma and Aspergillus sequences. Conclusions: Overall, the new discovery of common pathogens in agricultural field developed in live bed bugs storage tank may initiate the use of biological agents in later years

The development and validation of a scoring tool to predict the operative duration of elective laparoscopic cholecystectomy

Background: The ability to accurately predict operative duration has the potential to optimise theatre efficiency and utilisation, thus reducing costs and increasing staff and patient satisfaction. With laparoscopic cholecystectomy being one of the most commonly performed procedures worldwide, a tool to predict operative duration could be extremely beneficial to healthcare organisations. Methods: Data collected from the CholeS study on patients undergoing cholecystectomy in UK and Irish hospitals between 04/2014 and 05/2014 were used to study operative duration. A multivariable binary logistic regression model was produced in order to identify significant independent predictors of long (> 90 min) operations. The resulting model was converted to a risk score, which was subsequently validated on second cohort of patients using ROC curves. Results: After exclusions, data were available for 7227 patients in the derivation (CholeS) cohort. The median operative duration was 60 min (interquartile range 45–85), with 17.7% of operations lasting longer than 90 min. Ten factors were found to be significant independent predictors of operative durations > 90 min, including ASA, age, previous surgical admissions, BMI, gallbladder wall thickness and CBD diameter. A risk score was then produced from these factors, and applied to a cohort of 2405 patients from a tertiary centre for external validation. This returned an area under the ROC curve of 0.708 (SE = 0.013, p  90 min increasing more than eightfold from 5.1 to 41.8% in the extremes of the score. Conclusion: The scoring tool produced in this study was found to be significantly predictive of long operative durations on validation in an external cohort. As such, the tool may have the potential to enable organisations to better organise theatre lists and deliver greater efficiencies in care

IL-12 and IL-15 induce the expression of CXCR6 and CD49a on peripheral natural killer cells

Murine hepatic NK cells exhibit adaptive features, with liver-specific adhesion molecules CXCR6 and CD49a acting as surface markers. We investigated human liver-resident CXCR6+ and CD49a+ NK cells using RNA sequencing, flow cytometry and functional analysis. We further assessed the role of cytokines in generating NK cells with these phenotypes from the peripheral blood. Hepatic CD49a+ NK cells could be induced using cytokines and produce high quantities of IFNγ and TNFα, in contrast to hepatic CXCR6+ NK cells. RNA sequencing of liver-resident CXCR6+ NK cells confirmed a tolerant immature phenotype with reduced expression of markers associated with maturity and cytotoxicity. Liver-resident double positive CXCR6+CD49a+ hepatic NK cells are immature but maintain high expression of Th1 cytokines as observed for single-positive CD49a+ NK cells. We show that stimulation with activating cytokines can readily induce upregulation of both CD49a and CXCR6 on NK cells in the peripheral blood. In particular IL-12 and IL-15 can generate CXCR6+CD49a+ NK cells in vitro from NK cells isolated from the peripheral blood, with comparable phenotypic and functional features to liver-resident CD49a+ NK cells, including enhanced IFNγ and NKG2C expression. IL-12 and IL-15 may be key for generating NK cells with a tissue-homing phenotype and strong Th1 cytokine profile in the blood, and links peripheral activation of NK cells with tissue-homing. These findings may have important therapeutic implications for immunotherapy of chronic liver disease

Morphological and molecular characterization of fungus isolated from tropical bed bugs in Northern Peninsular Malaysia, Cimex hemipterus (Hemiptera: Cimicidae)

Objective: To investigate some morphological and molecular characteristics of fungal parasites isolated from wild tropical bed bug, Cimex hemipterus. Methods: A series of culture methods were used to obtain fungal isolates from field-collected bed bugs. Characteristics of the isolates such as colony appearance, mycelial texture and pigmentation were studied to explore their morphology. Isolates were also subjected to a PCR-based genotyping test. Results: There were noticeable differences in morphological characteristics among the four isolates. Conidial areas of one isolate were dark green, whereas those of the remaining colonies were olive-green, black or dark brown. Conidia of the dark green isolate were globose, while those of olive-green, black and dark brown isolates were globose to subglobose, globose to spherical and globose to subglobose/finely roughened, respectively. These morphological specificities and the molecular analyses showed that the fungal internal transcribed spacer ribosomal region and β-tubulin gene sequences of the isolates shared clade with Trichoderma and Aspergillus sequences. Conclusions: Overall, the new discovery of common pathogens in agricultural field developed in live bed bugs storage tank may initiate the use of biological agents in later years