Functional platelet-derived mitochondria induce the release of human neutrophil microvesicles.

Inflammation is an essential process of host defense against infections, illness, or tissue damage. Polymorphonuclear neutrophils (PMN) are among the first immune cells involved in acute inflammatory responses and are on the front line in the fight against bacterial infections. In the presence of bacterial fragments, PMN release inflammatory mediators, enzymes, and microvesicles in the extracellular milieu to recruit additional immune cells required to eliminate the pathogens. Recent evidence shows that platelets (PLTs), initially described for their role in coagulation, are involved in inflammatory responses. Furthermore, upon activation, PLT also release functional mitochondria (freeMitos) within their extracellular milieu. Mitochondria share characteristics with bacterial and mitochondrial damage-associated molecular patterns, which are important contributors in sterile inflammation processes. Deep sequencing transcriptome analysis demonstrates that freeMitos increase the mitochondrial gene expression in PMN. However, freeMitos do not affect the mitochondrial-dependent increase in oxygen consumption in PMN. Interestingly, freeMitos significantly induce the release of PMN-derived microvesicles. This study provides new insight into the role of freeMitos in the context of sterile inflammation.

Molecular characterization of the anticancer properties associated with bee venom and its components in glioblastoma multiforme.

Glioblastoma multiforme (GBM) is a frequent form of malignant glioma. Strategic therapeutic approaches to treat this type of brain tumor currently involves a combination of surgery, radiotherapy and chemotherapy. Nevertheless, survival of GBM patients remains in the 12-15 months range following diagnosis. Development of novel therapeutic approaches for this malignancy is therefore of utmost importance. Interestingly, bee venom and its components have shown promising anti-cancer activities in various types of cancer even though information pertaining to GBMs have been limited. The current work was thus undertaken to better characterize the anti-cancer properties of bee venom and its components in Hs683, T98G and U373 human glioma cells. MTT-based cell viability assays revealed IC50 values of 7.12, 15.35 and 7.60 µg/mL for cell lines Hs683, T98G and U373 treated with bee venom, respectively. Furthermore, melittin treatment of these cell lines resulted in IC50 values of 7.77, 31.53 and 12.34 µg/mL, respectively. Cell viability assessment by flow cytometry analysis confirmed signs of late apoptosis and necrosis after only 1 h of treatment with either bee venom or melittin in all three cell lines. Immunoblotting-based quantification of apoptotic markers demonstrated increased expression of Bak and Bax, while Caspsase-3 levels were significantly lower when compared to control cells. Quantification by qRT-PCR showed increased expression levels of long non-coding RNAs RP11-838N2.4 and XIST in glioma cells treated with either bee venom or melittin. Overall, this study provides preliminary insight on molecular mechanisms via which bee venom and its main components can impact viability of glioma cells and warrants further investigation of its anticancer potential in gliomas.

Evaluating the mitochondrial activity and inflammatory state of dimethyl sulfoxide differentiated PLB-985 cells.

Neutrophils play a key role in the innate immunity with their ability to generate and release inflammatory mediators that promote the inflammatory response and consequently restore the hemostasis. As active participants in several steps of the normal inflammatory response, neutrophils are also involved in chronic inflammatory diseases such as asthma, atherosclerosis, and arthritis. Given their dual role in the modulation of inflammation, regulating the inflammatory response of neutrophils has been suggested as an important therapeutic approach by numerous researchers. The neutrophils have a relatively short lifespan, which can be problematic for some in vitro experiments. To address this issue, researchers have used the human monomyelocyte cell line PLB-985 as an in vitro model for exploratory experiments addressing neutrophil-related physiological functions. PLB-985 cells can be differentiated into a neutrophil-like phenotype upon exposure to several agonists, including dimethyl sulfoxide (DMSO). Whether this differentiation of PLB-985 affects important features related to the neutrophil's normal functions (i.e., mitochondrial activity, eicosanoid production) remains elusive, and characterizing these changes will be the focal point of this study. Our results indicate that the differentiation affected the proliferation of PLB-985 cells, without inducing apoptosis. A significant decrease in mitochondrial respiration was observed in differentiated PLB-985 cells. However, the overall mitochondria content was not affected. Immunoblotting with mitochondrial antibodies revealed a strong modulation of the succinate dehydrogenase A, superoxide dismutase 2, ubiquinol-cytochrome c reductase core protein 2 and ATP synthase subunit α in differentiated PLB-985 cells. Finally, eicosanoids (leukotriene B4, 12-hydroxyheptadecatrienoic and 15-hydroxyeicosatetraenoic acids) production was significantly increased in differentiated cells. In summary, our data demonstrate that the differentiation process of PLB-985 cells does not impact their viability despite a reduced respiratory state of the cells. This process is also accompanied by modulation of the inflammatory state of the cell. Of importance, our data suggest that PLB-985 cells could be suitable in vitro candidates to study mitochondrial-related dysfunctions in inflammatory diseases.

Amyotrophic lateral sclerosis: update on clinical management.

PURPOSE OF REVIEW: The current review will provide recent updates in the clinical management of amyotrophic lateral sclerosis (ALS). RECENT FINDINGS: Although there is no cure for ALS, there are new treatments, growing knowledge of genetics, development of clinical staging systems, and the recent coronavirus disease 2019 pandemic that have recently impacted the clinical management of ALS. Increased understanding of genetics has helped provide insights into pathophysiology, the staging systems and clinical measures help to provide tools for monitoring disease clinically, and the recent coronavirus disease 2019 pandemic has provided opportunities to develop telemedicine and remote monitoring of disease thereby increasing accessibility to care and reducing burden of travel to centers for people living with the disease and their caregivers. SUMMARY: ALS is a progressive neurodegenerative disease that causes degeneration of the motor neurons which leads to paralysis and respiratory failure. Despite the lack of a cure, multidisciplinary care, proactive respiratory management, nutritional care and management of symptoms as well as pharmacological interventions that can improve quality of life and survival.

Defining the research agenda to measure and reduce tuberculosis stigmas.

Crucial to finding and treating the 4 million tuberculosis (TB) patients currently missed by national TB programmes, TB stigma is receiving well-deserved and long-delayed attention at the global level. However, the ability to measure and evaluate the success of TB stigma-reduction efforts is limited by the need for additional tools. At a 2016 TB stigma-measurement meeting held in The Hague, The Netherlands, stigma experts discussed and proposed a research agenda around four themes: 1) drivers: what are the main drivers and domains of TB stigma(s)?; 2) consequences: how consequential are TB stigmas and how are negative impacts most felt?; 3) burden: what is the global prevalence and distribution of TB stigma(s) and what explains any variation? 4): intervention: what can be done to reduce the extent and impact of TB stigma(s)? Each theme was further subdivided into research topics to be addressed to move the agenda forward. These include greater clarity on what causes TB stigmas to emerge and thrive, the difficulty of measuring the complexity of stigma, and the improbability of a universal stigma 'cure'. Nevertheless, these challenges should not hinder investments in the measurement and reduction of TB stigma. We believe it is time to focus on how, and not whether, the global community should measure and reduce TB stigma.

Diagnosis of Helicobacter pylori by carbon-13 urea breath test using a portable mass spectrometer.

CONTEXT: In the non-invasive detection of markers of disease, mass spectrometry is able to detect small quantities of volatile markers in exhaled air. However, the problem of size, expense and immobility of conventional mass spectrometry equipment has restricted its use. Now, a smaller, less expensive, portable quadrupole mass spectrometer system has been developed. Helicobacter pylori has been implicated in the development of chronic gastritis, gastric and duodenal ulcers and gastric cancer. OBJECTIVES: To compare the results obtained from the presence of H. pylori by a carbon-13 urea test using a portable quadrupole mass spectrometer system with those from a fixed mass spectrometer in a hospital-based clinical trial. METHODS: Following ethical approval, 45 patients attending a gastroenterology clinic at the Royal Liverpool University Hospital exhaled a breath sample into a Tedlar gas sampling bag. They then drank an orange juice containing urea radiolabelled with carbon and 30 min later gave a second breath sample. The carbon-13 content of both samples was measured using both quadrupole mass spectrometer systems. If the post-drink level exceeded the pre-drink level by 3% or more, a positive diagnosis for the presence of H. pylori was made. RESULTS: The findings were compared to the results using conventional isotope ratio mass spectrometry using a laboratory-based magnetic sector instrument off-site. The results showed agreement in 39 of the 45 patients. CONCLUSIONS: This study suggests that a portable quadrupole mass spectrometer is a potential alternative to the conventional centralised testing equipment. Future development of the portable quadrupole mass spectrometer to reduce further its size and cost is indicated, together with further work to validate this new equipment and to enhance its use in mass spectrometry diagnosis of other medical conditions.

New Zealand pathologists: a case study in occupational control.

This paper examines the progressive exertion of external managerial control over New Zealand pathologists as the country's New Public Management health reforms were implemented during the 1990s. Perspectives on professionalism, and its role in the effective use of resources, are discussed as part of the examination of this shift in decision-making power from pathologists to external management. Our analysis, based on a range of archived and interview data collected over the period 1997-2000, suggests that publicly unacceptable compromises in pathology service quality were risked by the pursuit of tight bureaucratic and free market controls over pathology practice. The paper concludes with suggestions for a health professional control model facilitative of maximal health gain.

Contribution of dialysis to endogenous oxalate production in patients with chronic renal failure.

We tested the possibility that the buffering agents in dialysis bath fluid might contribute to increased endogenous oxalate production in dialyzed patients. Using stable isotope dilution mass spectrometry, we obtained oxalate production rates and pool sizes directly for 10 patients in chronic renal failure, 5 of whom were undergoing continuous ambulatory peritoneal dialysis (lactate-buffered fluid). All peritoneal dialysis patients had either increased oxalate production rates or expanded oxalate pools when compared with undialyzed patients in renal failure. From a further four patients receiving maintenance hemodialysis we took blood samples immediately before and after three consecutive dialysis sessions in which the bath-fluid buffering agent (bicarbonate or acetate) was alternated; we analyzed these samples for oxalate and key precursors by capillary gas chromatography. Plasma glycine and serine concentrations remained within the physiological range. Glycolate and oxalate concentrations decreased, but the oxalate remained above normal after dialysis. All changes were independent of the bath-fluid buffering agent. We suggest that dialysis might stimulate the formation of oxalate by removing product inhibition of a late catabolic step.

The child's perception of the human energy field using Therapeutic Touch.

This is a descriptive, exploratory, phenomenological study on children's lived experiences of perceiving the human energy field using Therapeutic Touch (TT). Eleven children, 3 to 9 years of age, willingly participated in the study. The study's findings suggest that children can feel the human energy field with purpose or intent to help, thus supporting TT as an innate potential.

Decrease in xenon clearance during response to cold, dry air: problems of interpretation.

In order to increase our understanding of the nasal response to cold, dry air (CDA), we studied changes in xenon clearance as an indicator of nasal blood flow. Eight individuals previously shown to respond to CDA had measurements of xenon clearance made in the left inferior turbinate before, during, and after a 15-minute exposure to either CDA (-7 degrees C to 0 degrees C, less than 10% relative humidity) or room air. The half-life in seconds for xenon clearance on the day when CDA was inhaled was 56 +/- 6, 41 +/- 5, and 110 +/- 31, before, during, and 10 minutes after challenge, respectively. On the control day, with subjects breathing room air, the equivalent measurements of half-life in seconds were 54 +/- 8, 41 +/- 6, and 42 +/- 4, respectively. Xenon clearance was prolonged significantly (p less than .01) after exposure to CDA during the clinical response. The interpretation of the change in xenon clearance as an indicator of nasal blood flow is discussed.

CT of severe renal trauma in children: evaluation and course of healing with conservative therapy.

Over a 2-year period, blunt renal injuries were classified on a four-point scale: grade 1, contusions; grade 2, tears limited to the cortex (renal lacerations); grade 3, tears extending to the collecting system (renal fractures); and grade 4, renal vascular pedicle injuries. We report our findings in nine children with grade 2 and grade 3 blunt renal injuries who were evaluated with CT. One patient had a nephrectomy; the other eight were managed nonsurgically. Six patients had follow-up CT scans 5-19 months later to assess healing. Scars were evident in each case, and the extent of deformity paralleled the magnitude of the initial injury. One patient with a grade 2 injury and two patients with grade 3 injuries healed with small focal scars; three patients with grade 3 injuries healed with large polar scars. In five patients, the CT findings were compared with the findings on 99mTc-DTPA renal imaging. The injured kidneys contributed 30-45% (mean, 38%) of total renal function. In six patients with renal trauma who were treated conservatively, the involved kidneys healed and significant kidney function was preserved, although early surgical intervention might have been beneficial for one of these patients. Prospective studies are needed to evaluate further the effectiveness of this conservative approach.

Measurement of plasma oxalate by capillary gas chromatography and its validation by isotope dilution mass spectrometry.

A capillary gas chromatographic method for plasma oxalate and an isotope dilution mass spectrometric reference method, both using the same tert.-butyldimethylsilyl derivatives, are described. Similar reference ranges for both were found (4.93 +/- 1.48 and 4.70 +/- 1.44 mumol/l, respectively), together with a close correlation for results covering a wide range of oxalate concentrations.

Familial generalised resistance to thyroxine: case study.

We describe the clinical features of members of a family with an atypical form of thyroid hormone resistance syndrome, affecting thyroxine predominantly. Relevant diagnostic clinical and biochemical investigations are outlined and discussed.

An inherited albumin variant with enhanced affinity for the Amerlex thyroxin-analog.

We describe two siblings with artefactually increased results for free thyroxin in serum as measured with the Amerlex analog method, despite normal thyroxin transport. The cause of the artefact is identified as a variant albumin with enhanced affinity for the Amerlex thyroxin-analog.