Developing and Investigating a Nanovibration Intervention for the Prevention/Reversal of Bone Loss Following Spinal Cord Injury.

Osteoporosis disrupts the fine-tuned balance between bone formation and resorption, leading to reductions in bone quantity and quality and ultimately increasing fracture risk. Prevention and treatment of osteoporotic fractures is essential for reductions in mortality, morbidity, and the economic burden, particularly considering the aging global population. Extreme bone loss that mimics time-accelerated osteoporosis develops in the paralyzed limbs following complete spinal cord injury (SCI). In vitro nanoscale vibration (1 kHz, 30 or 90 nm amplitude) has been shown to drive differentiation of mesenchymal stem cells toward osteoblast-like phenotypes, enhancing osteogenesis and inhibiting osteoclastogenesis simultaneously. Here, we develop and characterize a wearable device designed to deliver and monitor continuous nanoamplitude vibration to the hindlimb long bones of rats with complete SCI. We investigate whether a clinically feasible dose of nanovibration (two 2 h/day, 5 days/week for 6 weeks) is effective at reversing the established SCI-induced osteoporosis. Laser interferometry and finite element analysis confirmed transmission of nanovibration into the bone, and microcomputed tomography and serum bone formation and resorption markers assessed effectiveness. The intervention did not reverse SCI-induced osteoporosis. However, serum analysis indicated an elevated concentration of the bone formation marker procollagen type 1 N-terminal propeptide (P1NP) in rats receiving 40 nm amplitude nanovibration, suggesting increased synthesis of type 1 collagen, the major organic component of bone. Therefore, enhanced doses of nanovibrational stimulus may yet prove beneficial in attenuating/reversing osteoporosis, particularly in less severe forms of osteoporosis.

Specialized DNA Structures Act as Genomic Beacons for Integration by Evolutionarily Diverse Retroviruses.

Retroviral integration site targeting is not random and plays a critical role in expression and long-term survival of the integrated provirus. To better understand the genomic environment surrounding retroviral integration sites, we performed a meta-analysis of previously published integration site data from evolutionarily diverse retroviruses, including new experimental data from HIV-1 subtypes A, B, C and D. We show here that evolutionarily divergent retroviruses exhibit distinct integration site profiles with strong preferences for integration near non-canonical B-form DNA (non-B DNA). We also show that in vivo-derived HIV-1 integration sites are significantly more enriched in transcriptionally silent regions and transcription-silencing non-B DNA features of the genome compared to in vitro-derived HIV-1 integration sites. Integration sites from individuals infected with HIV-1 subtype A, B, C or D viruses exhibited different preferences for common genomic and non-B DNA features. In addition, we identified several integration site hotspots shared between different HIV-1 subtypes, all of which were located in the non-B DNA feature slipped DNA. Together, these data show that although evolutionarily divergent retroviruses exhibit distinct integration site profiles, they all target non-B DNA for integration. These findings provide new insight into how retroviruses integrate into genomes for long-term survival.

Differential diagnosis of acute traumatic hip pain in the elderly.

Elderly patients who present with an inability to weight bear following a fall, with normal radiographs, should be appropriately investigated to rule out an occult hip fracture (OHF). We aim to identify both the range and incidence of the differential diagnosis of acute traumatic hip pain in a large series of patients investigated for OHF. A retrospective analysis of consecutive patients investigated for an OHF with magnetic resonance imaging (MRI) was performed. Dedicated musculo- skeletal radiologists reported the MRI scans. All diagnoses including hip fractures, other fractures and soft tissue injuries were recorded. Case notes were reviewed for all patients to identify subsequent complications, management and outcomes. A total of 157 patients fulfilled the inclusion criteria. 52 (33%) patients had a fracture of the proximal femur. The majority of patients with proximal femoral fractures required surgical intervention. 9 patients who had fractures of the greater trochanter of the femur without fracture extension across the femoral neck were managed non-operatively. 40 (25%) patients had fractures of the pelvis, with a combined pubic rami and sacral fracture occurring frequently. The most common diagnosis was a soft tissue injury alone that was seen in 60 (38%) patients imaged. Injuries to the gluteal muscle group, iliopsaos complex and trochanteric bursa were most prevalent. All patients with soft tissue injuries or fractures of the pelvis were successfully managed non-operatively. This study highlights a wide range of differential diagnoses in elderly patients presenting with acute traumatic hip pain. The proximal femur was fractured in 33% of patients imaged for OHFs in our series. The most common diagnosis was a soft tissue injury around the hip and pelvis ; these injuries can be successfully managed without surgery.

Treating COVID-19 Acute Severe Hypoxemic Respiratory Failure: First Case Report Utilizing Dexamethasone, Remdesivir, and Convalescent Plasma in Operation Inherent Resolve.

Coronavirus 2019 (COVID-19) has spread across the globe with a concerningly high infectivity resulting in the World Health Organization deeming it a pandemic. It has resulted in thousands of deaths and placed enormous strain on communities, healthcare systems and healthcare workers as they battle shortages of ventilators, supplies, and difficulties in protecting patients and hospital staff alike. Challenges in managing the disease have led to new treatment and management strategies as healthcare teams struggle to adapt. We present the first case of COVID-19 managed in the austere deployed environment of Operation Inherent Resolve in which the patient was treated with dexamethasone, remdesivir, COVID-19 convalescent plasma, positive pressure ventilation, and proning. We discuss some of the inherent and unique challenges of caring for a patient in this resource constrained environment with a brief review of the literature on the treatment and management.

Novel high throughput pooled shRNA screening identifies NQO1 as a potential drug target for host directed therapy for tuberculosis.

UNLABELLED: Chemical regulation of macrophage function is one key strategy for developing host-directed adjuvant therapies for tuberculosis (TB). A critical step to develop these therapies is the identification and characterization of specific macrophage molecules and pathways with a high potential to serve as drug targets. Using a barcoded lentivirus-based pooled short-hairpin RNA (shRNA) library combined with next generation sequencing, we identified 205 silenced host genes highly enriched in mycobacteria-resistant macrophages. Twenty-one of these "hits" belonged to the oxidoreductase functional category. NAD(P)H: quinone oxidoreductase 1 (NQO1) was the top oxidoreductase "hit". NQO1 expression was increased after mycobacterial infection, and NQO1 knockdown increased macrophage differentiation, NF-κB activation, and the secretion of pro-inflammatory cytokines TNF-α and IL-1ß in response to infection. This suggests that mycobacteria hijacks NQO1 to down-regulate pro-inflammatory and anti-bacterial functions. The competitive inhibitor of NQO1 dicoumarol synergized with rifampin to promote intracellular killing of mycobacteria. Thus, NQO1 is a new host target in mycobacterial infection that could potentially be exploited to increase antibiotic efficacy in vivo. Our findings also suggest that pooled shRNA libraries could be valuable tools for genome-wide screening in the search for novel druggable host targets for adjunctive TB therapies.

Identification of variants in primary and recurrent glioblastoma using a cancer-specific gene panel and whole exome sequencing.

Glioblastoma (GBM) is an aggressive, malignant brain tumor typically resulting in death of the patient within one year following diagnosis; and those who survive beyond this point usually present with tumor recurrence within two years (5-year survival is 5%). The genetic heterogeneity of GBM has made the molecular characterization of these tumors an area of great interest and has led to identification of molecular subtypes in GBM. The availability of sequencing platforms that are both fast and economical can further the adoption of tumor sequencing in the clinical environment, potentially leading to identification of clinically actionable genetic targets. In this pilot study, comprised of triplet samples of normal blood, primary tumor, and recurrent tumor samples from three patients; we compared the ability of Illumina whole exome sequencing (ExomeSeq) and the Ion AmpliSeq Comprehensive Cancer Panel (CCP) to identify somatic variants in patient-paired primary and recurrent tumor samples. Thirteen genes were found to harbor variants, the majority of which were exclusive to the ExomeSeq data. Surprisingly, only two variants were identified by both platforms and they were located within the PTCH1 and NF1 genes. Although preliminary in nature, this work highlights major differences in variant identification in data generated from the two platforms. Additional studies with larger samples sizes are needed to further explore the differences between these technologies and to enhance our understanding of the clinical utility of panel based platforms in genomic profiling of brain tumors.

Method development and validation for the simultaneous determination of organochlorine and organophosphorus pesticides in a complex sediment matrix.

The Xochimilco area in the southeastern part of Mexico City has a variety of socioeconomic activities, such as periurban agriculture, which is of great importance in the Mexico City metropolitan area. Pesticides are used extensively, some being legal, mostly chlorpyrifos and malathion, and some illegal, mostly DDT. Sediments are a common sink for pesticides in aquatic systems near agricultural areas, and Xochimilco sediments have a complex composition with high contents of organic matter and clay that are ideal adsorption sites for organochlorine (OC) and organophosphorus (OP) pesticides. Therefore, it is important to have a quick, affordable, and reliable method to determine these pesticides. Conventional methods for the determination of OC and OP pesticides are long, laborious, and costly owing to the high volume of solvents and adsorbents. The present study developed and validated a method for determining 18 OC and five OP pesticides in sediments with high organic and clay contents. In contrast with other methods described in the literature, this method allows isolation of the 23 pesticides with a 12 min microwave-assisted extraction (MAE) and one-step cleanup of pesticides. The method developed is a simpler, time-saving procedure that uses only 3.5 g of dry sediment. The use of MAE eliminates excessive handling and the possible loss of analytes. It was shown that the use of LC-Si cartridges with hexane-ethyl acetate (75+25, v/v) in the cleanup procedure recovered all pesticides with rates between 70 and 120%. The validation parameters demonstrated good performance of the method, with intermediate precision ranging from 7.3 to 17.0%, HorRat indexes all below 0.5, and tests of accuracy with the 23 pesticides at three concentration levels demonstrating recoveries ranging from 74 to 114% and RSDs from 3.3 to 12.7%.

Memory complaint questionnaire performed poorly as screening tool: validation against psychometric tests and affective measures.

OBJECTIVE: This study examined the internal and external validity of the Memory Complaint Questionnaire (MAC-Q), a brief measure of subjective memory complaint in people with normal cognitive function. STUDY DESIGN AND SETTING: The Study of Health Outcomes in Aircraft Maintenance Personnel was a retrospective cohort study investigating the association between aircraft fuel tank deseal/reseal activities and health status in Royal Australian Air Force personnel. Cross-sectional comparison tests included measures of executive functioning, psychomotor speed, attention/working memory, new learning/memory, depression, and anxiety. An adjusted regression analysis accounted for confounders including age, dates of posting, rank, education, alcohol use, tobacco use, and affective status. RESULTS: Eight hundred seventy-nine participants completed the MAC-Q. Although the MAC-Q tested as highly reliable and internally valid, it was highly associated with affective status and was only associated with Digit Symbol Coding after adjustment for depression/anxiety. CONCLUSION: The MAC-Q is greatly influenced by affective status but not memory performance. It is probably not useful as a specific screen of memory complaint for general population research.

Hearing impairment in F-111 maintenance workers: the study of health outcomes in aircraft maintenance personnel (SHOAMP) general health and medical study.

BACKGROUND: We sought to examine hearing loss in a group from the Royal Australian Air Force who undertook fuel tank maintenance on F-111 aircraft, with exposure to formulations containing ototoxins, relative to two different comparison groups. METHODS: Using pure-tone audiometry, hearing thresholds were assessed in 614 exposed personnel, 513 technical-trade comparisons (different base, same job), and 403 non-technical comparisons (same base, different job). We calculated percentage loss of hearing (PLH) and used regression models to examine whether there was an association between PLH and F-111 fuel tank maintenance, adjusting for possible confounders. In addition, the difference between the observed hearing thresholds and the expected thresholds based on an otologically normal population (ISO-7029-2003) was determined. RESULTS: The PLH ranged from nil to 96 (median 1.5, quartiles 0.3, 5.5). A logistic regression model showed no statistically significant difference in PLH among the three exposure groups (exposed vs. non-technical controls 1.1: 95% CI 0.7, 2.0 and exposed vs. technical OR 0.9: 95% CI 0.6, 1.3). The model also highlighted a number of other risk factors for PLH including age, tinnitus, smoking, depression, and use of depression medications. However, at all eight frequencies measured, all populations had lower than expected hearing thresholds based on published ISO-7029 medians. CONCLUSIONS: Although there was no difference in PLH between the three exposure groups, the study did reveal a high degree of hearing loss between the 3 groups and a normal population.

Is psychological distress in people living with cancer related to the fact of diagnosis, current treatment or level of disability? Findings from a large Australian study.

OBJECTIVE: To investigate whether the observed elevated levels of psychological distress in cancer survivors relate specifically to aspects of cancer diagnosis, to treatment or to disability. DESIGN, PARTICIPANTS AND SETTING: Self-reported questionnaire data on demographic, health and lifestyle factors and mental health from 89574 Australian men and women aged 45 years or older, sampled from the Medicare database for the 45 and Up Study from 1 February 2006 to 30 April 2008. Logistic regression was used to examine the risk of high levels of psychological distress in relation to cancer diagnosis and disability, adjusting for age, sex, income and education. MAIN OUTCOME MEASURE: High psychological distress (Kessler Psychological Distress Scale score > or = 22). RESULTS: Overall, 7.5% of participants had high levels of psychological distress. Among cancer survivors, the median time since diagnosis was 7.3 years. Compared with people without cancer, the odds ratios (95% CIs) for psychological distress were: 1.17 (1.09-1.26) in people reporting having had any cancer apart from non-melanoma skin cancer; 1.34 (1.08-1.67) in those with cancer diagnosed in the previous year; 1.53 (1.33-1.76) for those reporting treatment for cancer in the previous month and 1.11 (1.03-1.19) for those with cancer but without recent treatment. Using individuals with neither cancer nor disability as the reference group, the adjusted odds ratio (95% CI) for psychological distress was 6.51 (5.95-7.12) in those reporting significant disability but no cancer, 1.14(1.04-1.24) in those without disability but with cancer and 5.81(4.88-6.91) in those with both cancer and disability. CONCLUSION: The risk of psychological distress in individuals with cancer relates much more strongly to their level of disability than it does to the cancer diagnosis itself.

Environmental health impacts of equine estrogens derived from hormone replacement therapy.

Many factors have been considered in evaluations of the risk-benefit balance of hormone replacement therapy (HRT), used for treating menopausal symptoms in women, but not its potential risks for the environment We investigated the possible environmental health implications of conjugated equine estrogens (CEEs), the most common components of HRT, including their discharge into the environment, their uptake, potency, and ability to induce biological effects in wildlife. Influents and effluents from four U.K. sewage treatment works (STWs), and bile of effluent-exposed fish, were screened for six equine estrogens. In vitro estrogen receptor (ER) activation assays were applied in humans and fish to compare their potencies, followed by in vivo exposures of fish to equine estrogens and evaluation of bioaccumulation, estrogenic responses, and ER gene expression. The equine estrogen equilenin (Eqn), and its metabolite 17beta-dihydroequilenin (17beta-Eqn), were detected by tandem GC-MSMS in all STW influent samples and 83% of STW effluent samples analyzed, respectively, at low concentrations (0.07-2.6 ng/L) and were taken-up into effluent-exposed fish. As occurs in humans, these estrogens bound to and activated the fish ERs, with potencies at ERalpha 2.4-3490% of thatfor 17beta-estradiol. Exposure of fish for 21 days to Eqn and 17beta-Eqn induced estrogenic responses including hepatic growth and vitellogenin production at concentrations as low as 0.6-4.2 ng/L. Associated with these effects were inductions of hepatic ERalpha and ERbeta1 gene expression, suggesting ER-mediated mechanism(s) of action. These data provide evidence for the discharge of equine estrogens from HRT into the aquatic environment and highlight a strong likelihood that these compounds contribute to feminization in exposed wildlife.

Polychlorinated biphenyls in bile of patients with biliary tract cancer.

BACKGROUND: Polychlorinated biphenyls (PCBs) are anthropogenic, organic compounds. Although banned in the 1970s, PCBs are poorly biodegradable and hence ubiquitous in the environment. They accumulate in adipose tissue and are implicated various malignancies, including breast and pancreatic cancer. The hepatobiliary system is the main excretory route for such xenobiotic toxins. Incidence rates of intrahepatic biliary tract cancer are increasing worldwide. Measurement and comparison of PCB levels in bile from human patients with benign and malignant bile duct disease has not previously been done. OBJECTIVES: To compare PCB concentrations in bile from patients with malignant (n=8) and non-malignant (n=7) biliary disease. METHODS AND RESULTS: Fifteen human bile samples, collected endoscopically, were analysed using gas chromatography mass spectrometry for seven target PCB congeners (28, 52, 101, 118, 153, 138, and 180), known to occur in the environment and food. Amongst males, total PCB concentrations in bile ranged from 6 ng mL(-1) (aged 73 years) to 49 ng mL(-1) (aged 90 years); and in females between 8 ng mL(-1) (aged 33 years) to 43 ng mL(-1) (aged 67 years) bile. Although there was no overall difference in mean PCB levels between non-cancer and cancer patients, levels of congener 28 were significantly higher in patients with biliary tract cancer (p<0.05). CONCLUSIONS: Despite the banning of PCBs over 30 years ago, these xenobiotics are present in the bile of patients with biliary disease. PCB levels tend to increase with age, suggesting chronic bioaccumulation. Further research is necessary to investigate the relevance of increased levels of congener 28 in bile in biliary tract cancer.

The analysis of a group of acidic pharmaceuticals, carbamazepine, and potential endocrine disrupting compounds in wastewater irrigated soils by gas chromatography-mass spectrometry.

The analysis of pharmaceuticals and potential endocrine disruptors in the environment has rightly concentrated on their presence in wastewaters and possible contamination of receiving bodies, such as groundwaters. However, wastewater is increasingly being reused for irrigation and in order to fully understand the environmental fate of these compounds, reliable methods for their analysis in soil are required, of which there are relatively few available. This article reports a method for a range of acidic pharmaceuticals, carbamazepine, and endocrine disrupting compounds in soils with final analysis by gas chromatography-mass spectrometry. Two soil types (Phaeozom and Leptosol) and three fortification levels were used to validate the method. Recoveries of acidic pharmaceuticals varied between 62 and 102%, carbamazepine from 75 to 118%, and potential endocrine disruptors between 54 and 109%; most recoveries were between 75 and 95% and relative standard deviations were generally less than 10%. Detection limits were between 0.25 and 2.5 ng/g except for phthalates and 4-nonylphenols (25 ng/g). The method was used to analyze soils where untreated wastewaters have been used to irrigate crops for approximately 90 years. Concentrations of acidic pharmaceuticals in the soil were <1 ng/g and potential endocrine disruptors varied from below the limit of detection (estrone, 17beta-estradiol, and 17alpha-ethinylestradiol) to 2079 ng/L (bis-diethylhexyl phthalate). This data indicated that despite the continuous application of the contaminants over many years, concentrations were generally lower than those expected to be contributed by a single irrigation event. Only carbamazepine, at concentrations of 6.48 ng/g (in Phaeozem) and 5.14 ng/g (in Leptosol), showed any evidence of persistence in the soils analyzed.

Cancer incidence and mortality in aircraft maintenance workers.

BACKGROUND: A cancer incidence and mortality study was conducted in response to health concerns raised by workers from F-111 aircraft deseal/reseal fuel tank maintenance programs, to determine whether personnel exposed to deseal/reseal had an excess of cancers and mortality. METHODS: Number of deaths and cancers for individuals involved in F-111 DSRS activities were matched against two Air Force comparison groups. Analyses were weighted to adjust for differences in age, exposure period and rank. RESULTS: Eight hundred seventy-three exposed, 7,577 comparison group one, and 9,408 comparison group two individuals were matched against death and cancer data, with 431 cancers and 431 deaths. Cancer incidence was higher in the exposed group, with marginally significant increases of 40-50% (cancer incidence rate ratio range 1.45-1.62). Exposed group mortality was significantly lower than both comparison groups, likely due to survivor bias in the exposed group (mortality rate ratio range 0.33-0.44). CONCLUSIONS: On the balance of probabilities, there is an increased risk of cancer associated with participation in F-111 deseal/reseal activities.

Determination of acidic pharmaceuticals and potential endocrine disrupting compounds in wastewaters and spring waters by selective elution and analysis by gas chromatography-mass spectrometry.

Although the trend in development of analytical methods for emerging contaminants is towards reduced sample preparation and increased detector selectivity, there are still benefits from removal of matrix material during sample preparation. This paper describes a simple method for acidic pharmaceuticals and a range of potential endocrine disrupting compounds in untreated wastewaters and spring waters. It is based on separation of the two classes during elution from the extraction cartridge with final analysis by gas chromatography-mass spectrometry. 3,4-D was used as the recovery standard for the acids while 4-n-nonylphenol and [2H4]estrone were used for the endocrine disrupters; mean recoveries varied between 89% and 111%. The method was also extensively validated by fortification with the target compounds. Recoveries of acids were from 68% to 97% with relative standard deviations generally less than 10% and recoveries of endocrine disrupters were 68-109% with relative standard deviations less than 20%. Detection limits varied from 0.005 to 1 ng/L in spring water, and from 0.5 to 100 ng/L in untreated wastewater. Concentrations of the analytes in the wastewater ranged from 0.018 to 22.4 microg/L. Values were comparable to reported data, although concentrations were generally relatively high, probably because of a lack of treatment. Triclosan, phthalates, estrone, 17beta-estradiol, ibuprofen, and naproxen were present in the spring water from aquifers recharged indirectly with this wastewater after its use for irrigation; concentrations ranged from 0.01 to 25.0 ng/L. The much lower concentrations compared to wastewater indicate effective removal processes on passage through the soil and subsoil.

Continence promotion for older hospital patients following surgery for fractured neck of femur: pilot of a randomized controlled trial.

Evidence suggests that bladder control problems develop or worsen as a result of fractured neck of femur (#NOF) and its subsequent management. The primary aim of this study was to reduce the prevalence and severity of post surgery continence problems among patients, aged from 60-years, undergoing surgery for #NOF, using a best practice "case-management model" multifactorial intervention. Eligible consenting patients admitted with #NOF were randomized to intervention or control group. Self-report questionnaires compared pre-surgery, post surgery, and follow-up continence status between groups. This pilot randomized controlled trial, which included 45 eligible patients aged 60 to 93-years, found no evidence that the intervention was effective in reducing prevalence of post-surgery incontinence in this acute setting. Staff surveys highlighted the need for open communication between the research team and hospital staff. Unclear results were attributed to the small sample size. A central outcome was evidence that intervention to improve continence management for older people post-surgery is imperative. Focused assessment and treatment for those most at risk of incontinence after #NOF would be more acceptable to staff and a more efficient use of resources. A simple screening tool would ensure that those most at risk are detected, and targeted for care.

Trends in women's risk factors and chronic conditions: findings from the Australian Longitudinal Study on Women's Health.

Chronic diseases present a growing challenge to women's health. This paper presents data from the Australian Longitudinal Study on Women's Health to show prevalence and incidence among three cohorts of women of six chronic conditions: hypertension, heart disease, diabetes, asthma, osteoporosis and arthritis. It also examines the role of five important risk factors (body mass index, level of physical activity, smoking, alcohol consumption and level of education) on these chronic conditions. The most striking finding is that being overweight or obese is the most important risk factor for chronic disease for women in all three age groups.

Endogenous thrombin potential for predicting risk of venous thromboembolism in carriers of factor V Leiden.

Measurement of endogenous thrombin potential (ETP) detects hypercoagulability and can be used to identify activated protein C resistance due to factor V Leiden (FVL). However, not all carriers of FVL suffer thrombosis and therefore we sought to determine if the test for ETP could be modified in such a way as to enable detection of FVL patients who were at increased risk of venous thromboembolism. Protac, an activator of both protein C and factor V, was incorporated into the traditional thrombin generation reaction and ratios (reaction with Protac:reaction without Protac) were calculated. Plasma samples from 42 FVL heterozygotes (12 with a history of thrombosis and 30 with no prior thrombosis) and 38 controls (non-FVL with no history of thrombosis) were analysed. The mean ETP ratio was significantly higher in FVL heterozygotes (0.90 +/- 0.06) compared to normal controls (0.41 +/- 0.10; p = 0.00004). Multivariate analysis indicated that the average ETP ratio was significantly and inversely correlated with factor V levels in FVL heterozygotes (p = 0.002) but not controls. Within the FVL group, patients with a history of thrombosis had higher ETP ratios (0.92 +/- 0.06) compared to those without (0.89 +/- 0.05), however, this did not reach statistical significance (p = 0.09). Further investigation into the use of ETP for detecting risk of thrombosis in people who are genetically predisposed is warranted. The recent introduction of diagnostic ETP measurements in the form of the calibrated automated thrombin generation from Thrombinoscope and the TechnoThrombin from Baxter should facilitate such studies.