Standard Collection Methods Research Articles

Effectiveness of Biological and Surgical Sampling for Detection of Mycobacterium Tuberculosis

Introduction. Tuberculosis remains a significant public health issue, ranking second only to COVID-19 in terms of mortality from infectious diseases, with more than 10 million individuals affected annually. Despite a decline in incidence rate, its diagnosis continues to be challenging, particularly in cases with negative results from microbiological and histological examinations.Aim. To compare the effectiveness of surgical diagnostic methods for tuberculosis during the pre-analytical phase of bacteriological verification.Materials and methods. The retrospective study, conducted at the Republican Clinical Tuberculosis Treatment Dispensary of the Republic of Bashkortostan, analyzed data from 1272 patients who were hospitalized in the Tuberculous Pulmonary Surgery Unit from 2021 to 2023 and underwent bacteriological diagnostics, including microscopy and polymerase chain reaction.Results and discussion. The results indicate that out of 1272 surgeries, 65.2% involved resection procedures, with confirmed tuberculosis infection in 83.1% of patients. Bronchoalveolar lavage identified mycobacteria in 153 patients (20.5%), whereas sputum collection revealed them in 106 patients. The new collection method, although less effective, demonstrated a detection rate of 15.7% through molecular genetic analysis.Conclusion. Surgical diagnostic methods for tuberculosis appeared to be effective, detecting mycobacterium tuberculosis in 83.1% of cases, surpassing the standard sputum collection method, which has a detection rate of 10.8% via molecular genetic analysis. Bronchoalveolar lavage proved to be more productive, yielding 20.5% positive results on a molecular genetic basis, 11% microscopically, and 8% through culture. Although the new method is less effective, it is recorded as being more accurate than the standard, with detection rates of 15.7% molecularly, 6.6% microscopically, and 5.2% through culture.

A fish community comparison upstream and downstream a high-head dam on the Upper Mississippi River

Abstract Standardized sampling is useful for detecting and monitoring variations in fish communities and the establishment and spread of invasive species. Lock and Dam 19, a high-head dam with a 10- meter hydraulic head, creates the largest impoundment on the Upper Mississippi River. Upstream, Pool 19 is characterized by static water levels and expansive macrophyte beds. Downstream, Pool 20 is characterized by lotic conditions and sparse vegetation. Lock and Dam 19 serves as barrier to upstream migration of fishes, particularly Bigheaded carp (Bighead Carp Hypophthalmichthys nobilis and Silver Carp H. molotrix). There is limited standardized fisheries data available for Pools 19 and 20, particularly in off-channel areas. In 2013 and 2014, we conducted two years of standardized electrofishing in Pools 19 and 20 to 1) detect a presence of adult or juvenile Bigheaded carp, 2) quantify differences in fish community structure (i.e., relative abundance of species) between Pools 19 and 20 and among strata (i.e., backwater, side channel, main channel, and impounded reaches), and 3) quantify hydrological differences (i.e., water quality variables) between Pools 19 and 20 and among strata. We detected two Bigheaded carp upstream Lock and Dam 19, the first to be documented using a standardized collection method. We used permutational multivariate analysis of variance using distance matrices (veganR package version 2.6-4) to determine that fish community structure differed significantly between pools (P< 0.01) and among strata (P< 0.01), except between main and side channel borders. We determined water quality variables differed significantly between the pools and among strata demonstrating strata proportional differences may drive the fish community differences between Pools 19 and 20. We have demonstrated that expanding standardized sampling efforts to Pools 19 and 20 is important to understand long-term changes in fish assemblages and the effects of Lock and Dam 19 on the ecology of the Upper Mississippi River.

The Great Lakes Winter Grab: Limnological data from a multi‐institutional winter sampling campaign on the Laurentian Great Lakes

AbstractInterest in winter limnology is growing rapidly, but progress is hindered by a shortage of standardized multivariate datasets on winter conditions. Addressing the winter data gap will enhance our understanding of winter ecosystem function and of lake response to environmental change. Here, we describe a dataset generated by a multi‐institutional winter sampling campaign across all five Laurentian Great Lakes and some of their connecting waters (the Great Lakes Winter Grab). The objective of Winter Grab was to characterize mid‐winter limnological conditions in the Great Lakes using standard sample collection and analysis methods. Nineteen research groups sampled 49 locations varying widely in depth and trophic status, collecting a range of limnological data. This dataset includes physical, chemical, and biological measurements. These data can be used to examine diverse aspects of Great Lakes ecosystems or integrated with winter observations from other lakes to improve understanding of winter limnology across different aquatic systems.

Report of a new species of sand fly, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), from Western Ghats, India

BackgroundWestern Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India.MethodsA comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis.ResultsPhlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species.ConclusionsThe Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly.Graphical abstract

MicroRNA as Sepsis Biomarkers: A Comprehensive Review.

Sepsis, a life-threatening condition caused by the body's dysregulated response to infection, presents a significant challenge in clinical management. Timely and accurate diagnosis is paramount for initiating appropriate interventions and improving patient outcomes. In recent years, there has been growing interest in identifying biomarkers that can aid in the early detection and prognostication of sepsis. MicroRNAs (miRNAs) have emerged as potential biomarkers for sepsis due to their involvement in the regulation of gene expression and their stability in various biological fluids, including blood. MiRNAs are small non-coding RNA molecules that play crucial roles in post-transcriptional gene regulation by binding to target messenger RNAs (mRNAs), leading to mRNA degradation or translational repression. The diagnostic and prognostic potential of miRNAs in sepsis stems from their ability to serve as sensitive and specific biomarkers reflective of the underlying pathophysiological processes. Compared to traditional biomarkers such as C-reactive protein (CRP) and procalcitonin (PCT), miRNAs offer several advantages, including their early and sustained elevation during sepsis, as well as their stability in stored samples, making them attractive candidates for clinical use. However, despite their promise, the clinical translation of miRNAs as sepsis biomarkers faces several challenges. These include the need for standardized sample collection and processing methods, the identification of optimal miRNA panels or signatures for differentiating sepsis from other inflammatory conditions, and the validation of findings across diverse patient populations and clinical settings. In conclusion, miRNAs hold great promise as diagnostic and prognostic biomarkers for sepsis, offering insights into the underlying molecular mechanisms and potential therapeutic targets. However, further research is needed to overcome existing challenges and realize the full clinical utility of miRNAs in improving sepsis outcomes.

SARS-CoV-2 Viral Load in the Nasopharynx at Time of First Infection Among Unvaccinated Individuals

SARS-CoV-2 viral load (VL) in the nasopharynx is difficult to quantify and standardize across settings, but it may inform transmission potential and disease severity. To characterize VL at COVID-19 diagnosis among previously uninfected and unvaccinated individuals by evaluating the association of demographic and clinical characteristics, viral variant, and trial with VL, as well as the ability of VL to predict severe disease. This secondary cross-protocol analysis used individual-level data from placebo recipients from 4 harmonized, phase 3 COVID-19 vaccine efficacy trials sponsored by Moderna, AstraZeneca, Janssen, and Novavax. Participants were SARS-CoV-2 negative at baseline and acquired COVID-19 during the blinded phase of the trials. The setting included the US, Brazil, South Africa, Colombia, Argentina, Peru, Chile, and Mexico; start dates were July 27, 2020, to December 27, 2020; data cutoff dates were March 26, 2021, to July 30, 2021. Statistical analysis was performed from November 2022 to June 2023. Linear regression was used to assess the association of demographic and clinical characteristics, viral variant, and trial with polymerase chain reaction-measured log10 VL in nasal and/or nasopharyngeal swabs taken at the time of COVID-19 diagnosis. Among 1667 participants studied (886 [53.1%] male; 995 [59.7%] enrolled in the US; mean [SD] age, 46.7 [14.7] years; 204 [12.2%] aged 65 years or older; 196 [11.8%] American Indian or Alaska Native, 150 [9%] Black or African American, 1112 [66.7%] White; 762 [45.7%] Hispanic or Latino), median (IQR) log10 VL at diagnosis was 6.18 (4.66-7.12) log10 copies/mL. Participant characteristics and viral variant explained only 5.9% of the variability in VL. The independent factor with the highest observed differences was trial: Janssen participants had 0.54 log10 copies/mL lower mean VL vs Moderna participants (95% CI, 0.20 to 0.87 log10 copies/mL lower). In the Janssen study, which captured the largest number of COVID-19 events and variants and used the most intensive post-COVID surveillance, neither VL at diagnosis nor averaged over days 1 to 28 post diagnosis was associated with COVID-19 severity. In this study of placebo recipients from 4 randomized phase 3 trials, high variability was observed in SARS-CoV-2 VL at the time of COVID-19 diagnosis, and only a fraction was explained by individual participant characteristics or viral variant. These results suggest challenges for future studies of interventions seeking to influence VL and elevates the importance of standardized methods for specimen collection and viral load quantitation.

Electric field-assisted dried blood spot sample preparation for analysis of steroids using LC–MS/MS

Despite being a minimally invasive sample source, dried blood spots (DBS) generally pose the challenge of efficient sample extraction for broad metabolomics applications. This is particularly true for the quantification of steroids using non-derivatized liquid chromatography tandem mass spectrometry assays. To address these limitations, we have demonstrated the use of electric field as a driver for sample preparation from DBS samples to simultaneously quantify testosterone (T), 17α-hydroxyprogesterone (17-OHP), progesterone (P), and cortisol (C), using both standard electroporation cuvettes, as well as a novel custom vertical electric field setup. Our findings, backed by computational modeling, show that a 10V DC application for 180 s can draw out twice the amount of the aforementioned steroids from both Whatman-903 and DMPK-C sample collection cards using our novel device when compared to standard solvent-based collection methods. This study not only introduces the use of electric field for sample preparation for metabolomics, but additionally introduces a novel device that eliminates the electric double layer effect in the process.

Peri- and post-operative outcomes of robot-assisted radical cystectomy after the implementation of the EAU guidelines recommendations for collecting and reporting complications at a high-volume referral center.

No studies relied on a standardized methodology to collect postoperative complications after robot-assisted radical cystectomy (RARC). The aim of our study was to evaluate peri- and post-operative outcomes of patients undergoing RARC adhering to the European Association of Urology (EAU) recommendations for reporting surgical outcomes and using a long postoperative follow-up. 246 patients who underwent RARC with intracorporal urinary diversion at a single tertiary referral center with a postoperative follow-up ≥ 1year for survivors. Postoperative outcomes were collected prospectively by interviews done by medical doctors. Complications were scored using the Clavien-Dindo classification (CD), grouped by type and severity (severe: CD score ≥ 3). We described peri- and post-operative outcomes and complication chronological distribution. Overall, 16 (6.5%) and 225 patients (91%) experienced intraoperative and postoperative complications, respectively. Moreover, 139 (57%) experienced severe complications. The most common any-grade and severe complications were infectious (72%) and genitourinary (35%), respectively. Overall, 52% of complications (358/682) occurred within 10days from surgery, and 51% of severe complications (106/207) occurred within 35days. However, 13% of complications (90/682) and 28% of severe complications (59/207) occurred 3months after surgery. The earliest complications were fever of unknown origins and paralytic ileus (median time-to-complication [mTTC]: 4days), the latest complications were urinary tract infection (mTTC: 40days) and hydronephrosis/ureteral obstruction (mTTC: 70days). The rate of postoperative complications after RARC is > 90% when a standardized collection method and a long follow-up is implemented. These results should be used to identify potential areas of improvement and for preoperative patient counseling.

Aquatic insects as bioindicators of stream water quality - a seasonal analysis on Western Ghats river, Muthirapuzha, in central Kerala, India

This study was conducted to assess the water quality of Muthirapuzha River, Idukki using aquatic insects as bioindicators. Insects were collected on a seasonal basis from February 2014 to January 2015 from 12 sampling stations. Insects were sampled using standard collection methods and were identified up to family level. A total of 3,278 individuals belonging to seven orders and 37 families were collected during the study period. The greatest number of taxa was represented by order Ephemetroptera during monsoon (27%) and post-monsoon (25%), while Diptera (22.7%) dominated the pre-monsoon season. Shannon Weiner diversity index, Simpson dominance index, and Margalef’s richness index was highest post-monsoon. The EPT score in Muthirapuzha was for normal waters, however, pre-monsoon values were lowest, indicating pollution load during this period. Hilsenhoff’s family biotic index (HFBI) was used to estimate the status of organic pollution along the river based on representative families of aquatic entomofauna; values were highest at pre-monsoon season. The overall organic water quality level in the Muthirapuzha was good to fair based on this study.

Neurofilaments in Sporadic and Familial Amyotrophic Lateral Sclerosis: A Systematic Review and Meta-Analysis.

Neurofilament proteins have been implicated to be altered in amyotrophic lateral sclerosis (ALS). The objectives of this study were to assess the diagnostic and prognostic utility of neurofilaments in ALS. Studies were conducted in electronic databases (PubMed/MEDLINE, Embase, Web of Science, and Cochrane CENTRAL) from inception to 17 August 2023, and investigated neurofilament light (NfL) or phosphorylated neurofilament heavy chain (pNfH) in ALS. The study design, enrolment criteria, neurofilament concentrations, test accuracy, relationship between neurofilaments in cerebrospinal fluid (CSF) and blood, and clinical outcome were recorded. The protocol was registered with PROSPERO, CRD42022376939. Sixty studies with 8801 participants were included. Both NfL and pNfH measured in CSF showed high sensitivity and specificity in distinguishing ALS from disease mimics. Both NfL and pNfH measured in CSF correlated with their corresponding levels in blood (plasma or serum); however, there were stronger correlations between CSF NfL and blood NfL. NfL measured in blood exhibited high sensitivity and specificity in distinguishing ALS from controls. Both higher levels of NfL and pNfH either measured in blood or CSF were correlated with more severe symptoms as assessed by the ALS Functional Rating Scale Revised score and with a faster disease progression rate; however, only blood NfL levels were associated with shorter survival. Both NfL and pNfH measured in CSF or blood show high diagnostic utility and association with ALS functional scores and disease progression, while CSF NfL correlates strongly with blood (either plasma or serum) and is also associated with survival, supporting its use in clinical diagnostics and prognosis. Future work must be conducted in a prospective manner with standardized bio-specimen collection methods and analytical platforms, further improvement in immunoassays for quantification of pNfH in blood, and the identification of cut-offs across the ALS spectrum and controls.

Life cycle environmental impacts of kelp aquaculture through harmonized recalculation of inventory data

As seaweed farming gains prominence in future blue economies, scientifically robust environmental evaluations are vital. Harmonizing life cycle assessment (LCA) studies provides nuanced insights, allowing generalizations and potentially more accurate results than individual studies. This study recalculates life cycle inventory (LCI) data to offer a comprehensive perspective on sugar kelp Saccharina latissima. The findings affirm and validate previous studies, emphasizing critical hotspots such as fuel use for boats at sea, impacts from the use of plastic ropes, buoys and metal components at sea, and electrical energy use in the hatchery. The overall environmental impacts of seaweed farming remain relatively low compared to other seafood and biomass sources. The study also highlights the importance of how fuel use is modelled for the outcome. While harmonization enhances certainty and facilitates robust comparisons, challenges arise from the lack of standardized methods for data collection and reporting, along with data gaps between studies. Addressing these limitations calls for standardized protocols and improved data sharing practices in the field.

Tear Samples for Protein Extraction: Comparative Analysis of Schirmer's Test Strip and Microcapillary Tube Methods.

Tear sampling is an attractive option for collecting biological samples in ophthalmology clinics, as it offers a non-invasive alternative to other invasive techniques. However, there are many tear sampling methods still in consideration. This study explores the suitability of Schirmer's test strip and microcapillary tube as reliable and satisfactory methods for tear sampling. Tear samples were collected from eight healthy volunteers using the standard Schirmer's test strip method with or without anesthesia and microcapillary tubes. The total tear protein concentrations were analyzed via spectrophotometry and bicinchoninic acid (BCA) protein assay. The protein profile was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal wetting length of Schirmer's strip and suitable buffer solutions were compared. Discomfort levels reported by participants and the ease of execution for ophthalmologists were also evaluated. Tear samples exhibited typical protein profiles as shown by SDS-PAGE.The mean total protein obtained from an optimum wetting length of 20 mm using Schirmer's strip without anesthesia in phosphate-buffered saline (PBS) yielded substantial quantities of protein as measured by nanophotometer (220.20 ± 67.43 µg) and the BCA protein assay (210.34 ± 59.46 µg). This method collected a significantly higher quantity of protein compared to the microcapillary tube method (p=0.004) which was much more difficult to standardize. The clinician found it harder to utilize microcapillary tubes, while participants experienced higher insecurity and less discomfort with the microcapillary tube method.PBS used during the tear protein extraction process eluted higher tear protein concentration than ammonium bicarbonate, although the difference was not statistically significant. Using anaesthesia did not ease the sampling procedure substantially and protein quantity was maintained. Good quality and quantity of protein from tear samples were extracted with the optimized procedure. Schirmer's strip test in the absence of local anesthesia provided a standard, convenient, and non-invasive method for tear collection.

Quantitative photography for rapid, reliable measurement of marine macro‐plastic pollution

Abstract Plastics are now ubiquitous in the environment and have been studied in wildlife and in ecosystems for more than 50 years. Measurement of size, shape and colour data for individual fragments of plastic is labour‐intensive, unreliable and prone to observer bias, particularly when it comes to assessment of colour, which relies on arbitrary and inconsistently defined colour categorisations. There is a clear need for a standard method for data collection on plastic pollution, particularly one that can be readily automated given the number of samples involved. This study describes a new method for standardised photography of marine plastics in the 1–100 mm size range (meso‐ and macro‐plastics), including colour correction to account for any image‐to‐image variation in lighting that may impact colour reproduction or apparent brightness. Automated image analysis is then applied to detect individual fragments of plastic for quantitative measurement of size, shape, and colour. The method was tested on 3793 fragments of debris ingested by Flesh‐footed Shearwaters (Ardenna carneipes) on Lord Howe Island, Australia, and compare results from photos taken in two separate locations using different equipment. Photos were acquired of up to 250 fragments at a time with a spatial resolution of 70 μm/pixel and were colour‐corrected using a reference chart to ensure accurate reproduction of colour. The automated image analysis pipeline was found to have a 98% success rate at detecting fragments, and the different size and shape parameters that can be outputted by the pipeline were compared in terms of usefulness. The evidence shown in this study should strongly encourage the uptake of this method for cataloguing macro‐scale plastic pollution, as it provides substantially higher quality data with accurate, reliable measurements of size, shape and colour for individual plastics that can be readily compared between disparate datasets.

Optimization of a rapid method for screening drugs in blood by liquid chromatography tandem mass spectrometry.

In the recent years, liquid chromatography with tandem mass spectrometry has gained popularity in laboratories. This technique has a higher specificity, detects different analytes from a single specimen, measures analytes in distinct matrices, and substantially reduce analytical interference, with respect to immunoassay. The processing and preparation of biological samples are crucial in chromatography. Interferences in blood testing are usually caused by the presence of phospholipids and proteins. The main objective of this study was to improve analytical processes for drug screening by LC-MS/MS using a novel blood sample preparation method based on protein precipitation and removal of phospholipids. An evaluation was performed of a new method for the preparation of blood samples based on protein precipitation and removal of phospholipids by LC-Q-q-LIT. Limit of detection, limit of quantification and measurement range were determined for 56 molecules. The results of 11 cases were compared with those obtained using standard blood collection methods and instruments. The novel blood preparation and testing method based on LC-Q-q-LIT, a more sensitive technique, has demonstrated to yield comparable results to traditional methods. In addition, this new technique reduces turnaround time and costs.

Molecular Identification of Malaria Vector in Gusau Township, Nigeria

There about 460 Anopheles species recognized, over 100 can transmit human malaria but only 30–40 commonly transmit parasites of the genus Plasmodium, which cause malaria. Anopheles gambiae is the principal vector of the most dangerous malaria parasite species in Africa, which is Plasmodium falciparum. Anopheles gambiae are a complex consisting of eight morphologically indistinguishable species and each of the members of the complex having unique biology, ecology and behaviour and should be studied and differentiated. The research objective was to identify the species of malaria vector in Gusau township Nigeria using molecular biological technique to know the exact malaria vector. Knowledge of the exact malaria vector in a given environment enables formulating a carefully designed tailor-made vector control measure. Indoor and outdoor mosquito samples were collected from selected areas in five wards of Gusau township using standard collection methods, for a period of twelve months. The samples were identified using polymerase chain reaction (PCR) technique and the result showed Anopheles gambiae in all sites of sample collection. It is therefore recommended that integrated malaria vector control should be adopted in the mosquito control programme because Anopheles gambiae is rugged and can be difficult to control and/or eradicate because of the emerging insecticide resistance and its close association with human host.

PO-03-158 PEDIATRIC CARDIAC SCREENING DATA WAREHOUSE: A NATIONAL PUBLIC HEALTH RESOURCE

Cardiac screening of youth for prevention of sudden cardiac death in the young (SCDY) has been debated due to the absence of large population-specific screening data with outcomes. Despite years of screening by US public screening groups (PSG), there is minimal coordination of effort and no standardized methods for real-world data collection. To understand the methods, quality, outcomes, and best practices of youth screening, the Cardiac Safety Research Consortium Pediatric Cardiology Working Group, in collaboration with FDA and PSGs, developed and enabled a scalable system to collect a uniform pediatric cardiac screening dataset including digital ECGs and post-screening electronic follow-up data. Front end data collection (figure) was developed to include use of a universal unique ID system to align paper/digital collection of health and ECG data. PSGs use secure data transfer portals for digital ECG data upload for conversion to device-agnostic standardized FDA format to store in the national pediatric cardiac screening data warehouse. Follow-up data are obtained at designated post-screening intervals (one week, one and 3 months for pilot study) using initial text message contact followed by electronic consent (REDCap) and answering online health surveys. Fourteen PSGs in ten states participated in the pilot study. PSG warehouse data include 33840 retrospective ECG datasets collected from 2010 to 2021 containing limited screened history/symptoms but demographics similar to US census as follows: Age 13-30y, Male/Female 57/43%, Asian 6%, Black 19%, Native American <1%, Pacific Islander <1%, White 68%, Other 4%; Hispanic/Non-Hispanic 27%/79%. Individual PSG site demographics reflected local populations. Prospective data collection since 2021 include >4000 uniform screening datasets (age, sex, race, ethnicity, ht, wt, screening H&P, COVID history, medications, digital ECG with results, screening outcome, and, if applicable, ECHO results). Follow up participation allowing initial cellular contact was high (avg 73%, range 51-91%/screening). Establishment of a national pediatric cardiac data warehouse enables large-scale aggregation of pediatric cardiac screening information to address deficits in the understanding and prevention of SCDY. This large real-world dataset will help establish normative data for pediatric ECGs which can facilitate development of new diagnostic tools such as machine learning and support pediatric drug and device development.

INVESTIGATION OF N-ACYL HOMOSERINE LACTONE-BASED QUORUM-SENSING SYSTEM AND ALIGINATE LYASE ACTIVITY IN MARINE BACTERIAL SPECIES OF GRIMONTIA MARINA AS01 AND ALTEROMONAS MACLEODII AS02

Production, detection, and reaction to external signaling molecules are essential steps in quorum sensing (QS) process. Through the use of QS, bacterial communities may synchronize their responses to shifts in the density and diversity of their vicinal neighbors. QS also play an important role in regulating enzmyatic activities among marine bacteria. The aim of the present study was to detect and identify N- acyl homoserine lactones (AHLs) based QS signaling molecules and the possible influence on alginate lyase in marine bacterial isolates of Grimontia marina AS01 and Alteromonas macleodi AS02. Marine water samples were collected from Arabian Sea, Karchi Pakistan, following the standard collection methods. Bacterial strains were isolated and pure cultured using Zobell 2216 marine medium. Molecular identification was achieved based on 16S rRNA gene analysis. Screening for AHLs was achieved using Agrobacterium tumefaciens A136 as a biosensor. Based on 16S rRNA analysis, the bacterial strains were identified as Grimontia marina strain AS01 (OP143768) and Alteromonas macleodii strain AS02 (OP143769). Cross-feeding bioassay revealed the positive reactions for the production of AHLs. Reversed phase-TLC analysis showed the identification of C6-HSL produced by G. marina AS01 and 3OXO-C6-HSL by A. macleodii AS02 strain. Moreover, QS inhibitor AiiA protein reduced the production of alginate lyase in A. macleodi AS02, while no effect was observed in G. marina AS01. These results substantiate the involvement of QS system in regulating alginate lyase activity in A. macleodii AS02. QS in marine bacteria may involve in hydrolysis of complex organic matter in marine environment.

A non-invasive method for the detection of glucose in human exhaled breath by condensation collection coupled with ion chromatography

An analytical method for the detection of glucose in human exhaled breath by non-invasive condensation collection coupled with ion chromatography was developed. A self-designed exhaled breath condensation device was constructed, through which human exhaled breath was condensed and collected. And the glucose in collected human exhaled breath condensate (EBC) was analyzed by ion chromatography with a pulsed amperometric instrument. The standard EBC collection method was established, and the key factors such as cooling temperature and sampling flow rate during condensation collection were investigated deeply, and a good linear correlation between blood glucose levels and exhaled breath glucose levels was obtained. The intra-day precision was 3.37%, and the inter-day precision was 3.83%. Furthermore, EBC from healthy people and diabetic patients was collected in fasting state and after meal. We found the breath glucose level in healthy volunteers was 0.11–2.34 ng/L and that in diabetic patients was 0.23–135.92 ng/L. In after meal samples, the breath glucose level is 10–100 times higher than that of healthy subjects, which offering the prospect of a non-invasive approach to the monitoring of diabetes.

Challenges of Large-scale Patient-reported Outcome Measures Collection in a Multidisciplinary Limb Salvage Center.

Challenges of Large-scale Patient-reported Outcome Measures Collection in a Multidisciplinary Limb Salvage Center.

Design and Evaluation of a Computational Phenotype to Identify Patients With Metastatic Breast Cancer Within the Electronic Health Record.

Outcomes for patients with metastatic breast cancer (MBC) are continually improving as more effective treatments become available. Granular data sets of this unique population are lacking, and the standard method for data collection relies largely on chart review. Therefore, using electronic health records (EHR) collected at a tertiary hospital system, we developed and evaluated a computational phenotype designed to identify all patients with MBC, and we compared the effectiveness of this algorithm against the gold standard, clinical chart review. A cohort of patients with breast cancer were identified according to International Classification of Diseases codes, the institutional tumor registry, and SNOMED codes. Chart review was performed to determine whether distant metastases had occurred. We developed a computational phenotype, on the basis of SNOMED concept IDs, which was applied to the EHR to identify patients with MBC. Contingency tables were used to aggregate and compare results. A total of 1,741 patients with breast cancer were identified using data from International Classification of Diseases codes, the tumor registry, and/or SNOMED concept identifiers. Chart review of all patients classified each patient as having MBC (n = 416; 23.9%) versus not (n = 1,325; 75.9%). The final computational phenotype successfully classified 1,646 patients (95% accuracy; 82% sensitivity; 99% specificity). Hospital systems with robust EHRs and reliable mapping to SNOMED have the ability to use standard codes to derive computational phenotypes. These algorithms perform reasonably well and have the added ability to be run at disparate health care facilities. Better tooling to navigate the polyhierarchical structure of SNOMED ontology could yield better-performing computational phenotypes.