Frailty and risk of metabolic dysfunction-associated steatotic liver disease and other chronic liver diseases.

BACKGROUND & AIMS: Frailty is associated with multiple morbidities. However, its effect on chronic liver diseases remains largely unexplored. This study evaluated the association of frailty with the risk of incident metabolic dysfunction-associated steatotic liver disease (MASLD), cirrhosis, liver cancer, and liver-related mortality. METHODS: A total of 339,298 participants without prior liver diseases from the UK Biobank were included. Baseline frailty was assessed by using physical frailty and the frailty index, categorizing participants as nonfrail, prefrail, or frail. The primary outcome was MASLD, with secondary outcomes, including cirrhosis, liver cancer, and liver-related mortality, confirmed through hospital admission records and death registries. RESULTS: During a median follow-up of 11.6 years, 4,667 MASLD, 1,636 cirrhosis, 257 liver cancer, and 646 liver-related mortality cases were identified. After multivariable adjustment, the risk of MASLD was found to be higher in participants with prefrailty (physical frailty: HR = 1.66, 95% CI = 1.40-1.97; frailty index: HR = 2.01, 95% CI = 1.67-2.42) and frailty (physical frailty: HR = 3.32, 95% CI = 2.54-4.34; frailty index: HR = 4.54, 95% CI = 3.65-5.66) than in those with nonfrailty. Similar results were also observed for cirrhosis, liver cancer, and liver-related mortality. Additionally, the frail groups had a higher risk of MASLD, which was defined as magnetic resonance imaging-derived liver proton density fat fraction > 5%, than the nonfrail group (physical frailty: OR = 1.64, 95% CI = 1.32-2.04; frailty index: OR = 1.48, 95% CI = 1.30-1.68). CONCLUSIONS: Frailty was associated with an increased risk of chronic liver diseases. Public health strategies should target reducing chronic liver disease risk in frail individuals. IMPACT AND IMPLICATIONS: While frailty is common and associated with a poor prognosis in people with MASLD and advanced chronic liver diseases, its impact on the subsequent risk of these outcomes remains largely unexplored. Our study showed that frailty was associated with the increased risks of MASLD, cirrhosis, liver cancer, and liver-related mortality. This finding suggests that assessing frailty may help identify a high-risk population vulnerable to developing chronic liver diseases. Implementing strategies that target frailty could have major public health benefits for liver-related disease prevention.

Combined associations of visceral adipose tissue and adherence to a Mediterranean lifestyle with T2D and diabetic microvascular complications among individuals with prediabetes.

BACKGROUND: It's unclear if excess visceral adipose tissue (VAT) mass in individuals with prediabetes can be countered by adherence to a Mediterranean lifestyle (MEDLIFE). We aimed to examine VAT mass, MEDLIFE adherence, and their impact on type 2 diabetes (T2D) and diabetic microvascular complications (DMC) in individuals with prediabetes. METHODS: 11,267 individuals with prediabetes from the UK Biobank cohort were included. VAT mass was predicted using a non-linear model, and adherence to the MEDLIFE was evaluated using the 25-item MEDLIFE index, encompassing categories such as "Mediterranean food consumption," "Mediterranean dietary habits," and "Physical activity, rest, social habits, and conviviality." Both VAT and MEDLIFE were categorized into quartiles, resulting in 16 combinations. Incident cases of T2D and related DMC were identified through clinical records. Cox proportional-hazards regression models were employed to examine associations, adjusting for potential confounding factors. RESULTS: Over a median follow-up of 13.77 years, we observed 1408 incident cases of T2D and 714 cases of any DMC. High adherence to the MEDLIFE, compared to the lowest quartile, reduced a 16% risk of incident T2D (HR: 0.84, 95% CI: 0.71-0.98) and 31% for incident DMC (0.69, 0.56-0.86). Conversely, compared to the lowest quartile of VAT, the highest quartile increased the risk of T2D (5.95, 4.72-7.49) and incident any DMC (1.79, 1.36-2.35). We observed an inverse dose-response relationship between MEDLIFE and T2D/DMC, and a dose-response relationship between VAT and all outcomes (P for trend < 0.05). Restricted cubic spline analysis confirmed a nearly linear dose-response pattern across all associations. Compared to individuals with the lowest MEDLIFE quartile and highest VAT quartile, those with the lowest T2D risk had the lowest VAT and highest MEDLIFE (0.12, 0.08-0.19). High MEDLIFE was linked to reduced T2D risk across all VAT categories, except in those with the highest VAT quartile. Similar trends were seen for DMC. CONCLUSION: High adherence to MEDLIFE reduced T2D and MDC risk in individuals with prediabetes, while high VAT mass increases it, but MEDLIFE adherence may offset VAT's risk partly. The Mediterranean lifestyle's adaptability to diverse populations suggests promise for preventing T2D.

Impact of healthy lifestyle on the incidence and progression trajectory of mental disorders: A prospective study in the UK Biobank.

BACKGROUND: Healthier lifestyle decreased the risk of mental disorders (MDs) such as depression and anxiety. However, research on the effects of a comprehensive healthy lifestyle on their progression is lacking. METHODS: 385,704 individuals without baseline MDs from the UK Biobank cohort were included. A composite healthy lifestyle score was computed by assessing alcohol intake, smoking status, television viewing time, physical activity, sleep duration, fruit and vegetable intake, oily fish intake, red meat intake, and processed meat intake. Follow-up utilized hospital and death register records. Multistate model was used to examine the role of healthy lifestyle on the progression of specific MDs, while a piecewise Cox regression model was utilized to assess the influence of healthy lifestyle across various phases of disease progression. RESULTS: Higher lifestyle score reduced risks of transitions from baseline to anxiety and depression, as well as from anxiety and depression to comorbidity, with corresponding hazard ratios (HR) and 95 % confidence intervals (CI) of 0.94 (0.93, 0.95), 0.90 (0.89, 0.91), 0.94 (0.91, 0.98), and 0.95 (0.92, 0.98), respectively. Healthier lifestyle decreased the risk of transitioning from anxiety to comorbidity within 2 years post-diagnosis, with HR 0.93 (0.88, 0.98). Higher lifestyle scores at 2-4 years and 4-6 years post-depression onset were associated with reduced risk of comorbidity, with HR 0.93 (0.87, 0.99) and 0.92 (0.86, 0.99), respectively. LIMITATION: The generalizability to other ethnic groups is limited. CONCLUSION: This study observed a protective role of holistic healthy lifestyle in the trajectory of MDs and contributed to identifying critical progression windows.

[Resuscitative Endovascular Balloon Occlusion of the Aorta: Implementation and Nursing Care].

Uncontrolled hemorrhagic shock is the main cause of death in patients with traumatic injuries. Resuscitative endovascular balloon occlusion of the aorta (REBOA) is a rapidly performed and less-invasive treatment to stop bleeding. The REBOA balloon may be placed at the location of bleeding to achieve hemostasis. Prior to balloon placement, the indications for placement must be evaluated, including non-thoracic aortic trauma and non-traumatic patients with postpartum hemorrhage, and the appropriate sheath size (from 5 to 8 French) must be selected based on the bleeding site. As vascular injury of aortic dissection, rupture, or perforation may occur during the procedure, changes in hemodynamic parameters should be monitored. After balloon placement, ischemic complications due to blood flow occlusion such as lower extremity ischemia and acute renal failure should be tracked. After balloon removal, reperfusion injuries may occur, which can result in multiple organ failure, and should be observed closely. When caring for patients receiving REBOA, physicians should explain the procedure to their families to obtain informed consent. Also, nurses should prepare supplies and closely monitor changes in critical life signs to minimize the risks of hypotension, arrhythmia, and changes in consciousness during the procedure. After placement, the neurovascular and peripheral limbs "5P" (pain, pallor, paresthesia, pulselessness and paralysis) condition should be evaluated and recorded in detail. This treatment approach requires further study and research to assess the long-term impacts of placement and improve quality of care in these patients.

Sorption of Cellulases in Biofilm Enhances Cellulose Degradation by Bacillus subtilis.

Biofilm commonly forms on the surfaces of cellulosic biomass but its roles in cellulose degradation remain largely unexplored. We used Bacillus subtilis to study possible mechanisms and the contributions of two major biofilm components, extracellular polysaccharides (EPS) and TasA protein, to submerged biofilm formation on cellulose and its degradation. We found that biofilm produced by B. subtilis is able to absorb exogenous cellulase added to the culture medium and also retain self-produced cellulase within the biofilm matrix. The bacteria that produced more biofilm degraded more cellulose compared to strains that produced less biofilm. Knockout strains that lacked both EPS and TasA formed a smaller amount of submerged biofilm on cellulose than the wild-type strain and also degraded less cellulose. Imaging of biofilm on cellulose suggests that bacteria, cellulose, and cellulases form cellulolytic biofilm complexes that facilitate synergistic cellulose degradation. This study brings additional insight into the important functions of biofilm in cellulose degradation and could potentiate the development of biofilm-based technology to enhance biomass degradation for biofuel production.

[Project to Reduce Cases of Drug-Resistant Strain Colonization in the Intensive Care Unit Using a Chlorhexidine Gluconate Bath].

BACKGROUND: Drug-resistant strains of bacteria are associated with severe consequences such as bacteremia, shock, and death, and increase hospital stay durations and medical health expenses. Therefore, reducing the spread of drug-resistant strains is a priority concern. PURPOSE: This project was developed to reduce the number of colonization cases of drug-resistant strains and subsequently increase the quality of care provided in our intensive care unit. RESOLUTIONS: In this project, a chlorhexidine gluconate (CHG) bath standard protocol and CHG bath skill checklist were established, education and training courses were planned, a regular bed curtain replacement schedule and sink cleaning protocols were implemented, and regular audits were conducted. RESULTS: Immediately following project implementation (October - December 2018), the average monthly cases of Vancomycin-resistant enterococci colonization decreased from 6.08 to 4.33, and the average monthly cases of multi-drug resistant Acinebacter baumannii colonization decreased from 4.08 to 1.33. Furthermore, between January and July 2019, the average monthly cases of colonization for the abovementioned bacteria numbered 4 and 0.86, respectively, which met the level of reduction targeted in this project. CONCLUSIONS: The results of this project indicate that implementing CHG bed baths is effective in reducing the incidence of drug resistant strain colonization and rate of related infections in patients. CHG bed baths should be applied clinically to improve the quality of intensive care.

Distinguishing Anuran species by high-resolution melting analysis of the COI barcode (COI-HRM).

Taxonomic identification can be difficult when two or more species appear morphologically similar. DNA barcoding based on the sequence of the mitochondrial cytochrome c oxidase 1 gene (COI) is now widely used in identifying animal species. High-resolution melting analysis (HRM) provides an alternative method for detecting sequence variations among amplicons without having to perform DNA sequencing. The purpose of this study was to determine whether HRM of the COI barcode can be used to distinguish animal species. Using anurans as a model, we found distinct COI melting profiles among three congeners of both Lithobates spp. and Hyla spp. Sequence variations within species shifted the melting temperature of one or more melting domains slightly but do not affect the distinctness of the melting profiles for each species. An NMDS ordination plot comparing melting peak profiles among eight Anuran species showed overlapping profiles for Lithobates sphenocephala and Gastrophryne carolinensis. The COI amplicon for both species contained two melting domains with melting temperatures that were similar between the two species. The two species belong to two different families, highlighting the fact that COI melting profiles do not reveal phylogenetic relationships but simply reflect DNA sequence differences among stretches of DNA within amplicons. This study suggests that high-resolution melting analysis of COI barcodes (COI-HRM) may be useful as a simple and rapid method to distinguish animal species that appear morphologically similar.

[Prone Position and Nursing Care].

Acute respiratory distress syndrome (ARDS) is a life-threatening disease, as acute inflammation in the lungs typically leads to hypoxia and symptoms of dyspnea. The treatment modalities of ARDS include mechanical ventilation, corticosteroid, extracorporeal membranous oxygenation, inhaled nitrogen oxide, and the prone position. Among these, the prone position is supported by evidence showing significantly reduced mortality in patients that adopt this modality. Lying in the prone position reduces atelectatic lung volumes by recruitment of dependent parts and facilitates normal regulation of alveolar ventilation, giving better-matched ventilation perfusion and, thus, improved oxygenation. Lying in the prone position should be initiated as early as possible and may be implemented when there is limited improvement after 12-24 hours under mechanical ventilation and when the PaO2/FiO2 ratio is less than 150 mmHg. However, the prone position may not be appropriate for patients with increased intracranial pressure, hemodynamic instability, an open abdominal wound, or pregnancy. Prior to setting a patient in the prone position, fixation of tubings should be affirmed, and all tubings should be reexamined after each adjustment in position in order to ensure that they are free of twists and function well. In addition, caution should be given to the skin at pressure points to avoid pressure sores and foam dressings may be applied in advance for protection. This article summarizes the pathophysiology of ARDS, the principle of applying the prone position, and related indications, complications, and nursing care in order to give nurses more confidence in caring for patients using the prone position.

Rapid differentiation of bacterial communities using high resolution melting analysis.

Analysis of microbial communities is of broad interest in biology and high throughput sequencing is now the preferred method. However, some studies may not need the level of detail high throughput sequencing provides and its cost may limit the number of samples that can be sequenced. High resolution melting analysis (HRM) of 16S rRNA gene variable regions has been proposed as an efficient and low cost method to prioritize samples for sequencing but more specific primers are needed and its efficacy needs to be confirmed. We tested a more specific pair of primers and compared results concerning the structure of microbial communities in tadpole intestine and feces obtained using HRM, denaturing gradient gel electrophoresis (DGGE) and high-throughput sequencing performed in parallel. All three methods showed similar conclusions concerning the communities and revealed communities that differed among intestinal regions and feces. The improved HRM method targets a shorter amplicon in the V3 region of the 16S rRNA gene and uses non-degenerate primers, both of which increase the sensitivity of HRM. The HRM approach was shown to be as effective as DGGE for comparing microbial communities, is considerably easier to perform, and can be used to assess differences in microbial community structure among a large number of samples before committing to sequencing.

[New Treatment for Vascular Thrombosis Prevention in Patients With Atrial Fibrillation: Left Atrial Appendage Occlusion].

Atrial fibrillation (AF) is a common type of arrhythmia that increases significantly the risk of blood clots in the heart and of stroke. Therefore, stroke prevention is a key goal of AF treatment. In the past, patients were required to take anticoagulants for the remainder of their life, to regularly the monitor international normalized ratio (INR) of prothrombin time (PT), and to avoid possible negative interactions with various drugs and foods. Left atrial appendage occlusion (LAAO), a novel device and technique, was thus developed for AF patients with contraindications to anticoagulants and a high risk of bleeding. When using this technique, the occluder is placed on the left atrial appendage in order to effectively prevent blood stasis and thrombi accumulation. Transesophageal echocardiogram and computed tomography are conducted prior to the LAAO procedure, which is similar to the procedure used for cardiac catheterization. After the LAAO procedure, the patient remains in the intensive care unit (ICU), where vital signs, bleeding at the puncture site, and pericardial tamponade complications are monitored. Health education on daily activities, anticoagulant use, and regular follow-up should be given prior to hospital discharge. While LAAO may not reduce the incidence of stroke, the benefits of this procedure include a significant reduction in bleeding complications as compared to procedures that use oral anticoagulants. Further studies including long-term follow up and in-depth examinations of this procedure are necessary. The present article offers a reference for clinical staffs who are responsible for the care of patients treated using the LAAO procedure.

Complex carbohydrates reduce the frequency of antagonistic interactions among bacteria degrading cellulose and xylan.

Bacterial competition for resources is common in nature but positive interactions among bacteria are also evident. We speculate that the structural complexity of substrate might play a role in mediating bacterial interactions. We tested the hypothesis that the frequency of antagonistic interactions among lignocellulolytic bacteria is reduced when complex polysaccharide is the main carbon source compared to when a simple sugar such as glucose is available. Results using all possible pairwise interactions among 35 bacteria isolated from salt marsh detritus showed that the frequency of antagonistic interactions was significantly lower on carboxymethyl cellulose (CMC)-xylan medium (7.8%) than on glucose medium (15.5%). The two interaction networks were also different in their structures. Although 75 antagonistic interactions occurred on both media, there were 115 that occurred only on glucose and 20 only on CMC-xylan, indicating that some antagonistic interactions were substrate specific. We also found that the frequency of antagonism differed among phylogenetic groups. Gammaproteobacteria and Bacillus sp. were the most antagonistic and they tended to antagonize Bacteroidetes and Actinobacteria, the most susceptible groups. Results from the study suggest that substrate complexity affects how bacteria interact and that bacterial interactions in a community are dynamic as nutrient conditions change.

Synergistic growth in bacteria depends on substrate complexity.

Both positive and negative interactions among bacteria take place in the environment. We hypothesize that the complexity of the substrate affects the way bacteria interact with greater cooperation in the presence of recalcitrant substrate. We isolated lignocellulolytic bacteria from salt marsh detritus and compared the growth, metabolic activity and enzyme production of pure cultures to those of three-species mixed cultures in lignocellulose and glucose media. Synergistic growth was common in lignocellulose medium containing carboxyl methyl cellulose, xylan and lignin but absent in glucose medium. Bacterial synergism promoted metabolic activity in synergistic mixed cultures but not the maximal growth rate (µ). Bacterial synergism also promoted the production of ß-1,4-glucosidase but not the production of cellobiohydrolase or ß-1,4-xylosidase. Our results suggest that the chemical complexity of the substrate affects the way bacteria interact. While a complex substrate such as lignocellulose promotes positive interactions and synergistic growth, a labile substrate such as glucose promotes negative interactions and competition. Synergistic interactions among indigenous bacteria are suggested to be important in promoting lignocellulose degradation in the environment.

[Introduction of Hemodynamic Monitoring in Critical Care Units].

Hemodynamic monitoring is a very important treatment in intensive care units. Measurements taken during monitoring include pulmonary artery catheter (PAC), pulse-induced contour output (PiCCO), and non-invasive hemodynamic monitoring. PAC measures cardiopulmonary parameters using the thermodilution principle. PiCCO uses transpulmonary thermodilution and pulse contour analysis to measure cardiopulmonary parameters and extra-vascular lung water, to predict lung edema, and to differentiate between cardiogenic and non-cardiogenic respiratory failure. Non-invasive hemodynamic monitoring uses the thoracic electrical bioimpedance principle to measure electrical conductivity and then calculates stroke volume and cardiopulmonary parameters using the arrangement of red blood cells. The author is a nurse in an intensive care unit who is familiar with the various methods used in hemodynamic monitoring, with preparing the related devices, with briefing patients and family members prior to procedures, with related aseptic skills, with preventing complications during the insertion procedure, and with analyzing and interpreting those parameters accurately. The issues addressed in this paper are provided as a reference for nurses and other medical personnel to choose appropriate treatments when caring for critical patients.

[Transcatheter Aortic Valve Implantation: An Introduction and Patient Care].

Aortic stenosis has a high prevalence among individuals over 75 years of age. Transcatheter aortic valve implantation (TAVI) is a novel valve-replacement technique for patients with multiple chronic diseases who are at high risk of requiring aortic valve replacement surgery. Most of the time, the indicators of TAVI are detected during an echocardiographic exam. The femoral artery is the primary insertion site. The complications of TAVI include stroke, vascular dissection, bleeding, aortic valve regurgitation, and arrhythmia. In terms of clinical effectiveness, the mortality rate of TAVI is lower than percutaneous ballon valvuloplasty but similar to AVR. The unplanned cardiac-related re-admission rate within 30 days of discharge is lower for TAVI than for AVR. In terms of activity tolerance, TAVI is significantly better than both percutaneous ballon valvuloplasty and AVR. Comprehensive nursing care may reduce the incidence of complications associated with TAVI. Nursing care of TAVI includes explaining and providing instructions regarding TAVI prior to the procedure. After the TAVI procedure and while the patient is in the ICU, remove the endotracheal tube as soon as possible, monitor his / her neuro-cognitive status, monitor for early detection of a stroke event, record urine output to assess renal function, observe bleeding in the puncture site, and evaluate cardiac arrhythmia and pain. While in the general ward, resume early physical activities and educate the patient regarding the risks and the prevention of bleeding. This article provides references for clinical staff responsible to care for post-TAVI surgery patients.

The ocean sampling day consortium.

Ocean Sampling Day was initiated by the EU-funded Micro B3 (Marine Microbial Biodiversity, Bioinformatics, Biotechnology) project to obtain a snapshot of the marine microbial biodiversity and function of the world's oceans. It is a simultaneous global mega-sequencing campaign aiming to generate the largest standardized microbial data set in a single day. This will be achievable only through the coordinated efforts of an Ocean Sampling Day Consortium, supportive partnerships and networks between sites. This commentary outlines the establishment, function and aims of the Consortium and describes our vision for a sustainable study of marine microbial communities and their embedded functional traits.

Application of mass spectrometry in newborn screening: about both small molecular diseases and lysosomal storage diseases.

Many genetic diseases, especially the inborn errors of metabolism, have very low incidences, so developing a newborn screening test for each disease is not practical. This obstacle was overcome by employing the tandem mass spectrometry (MS/MS) technology. In the analysis, the samples can be injected directly into the flowing system without passing through a column, and both acylcarnitine and amino acid profiles can be obtained at the same time. MS/MS newborn screening has been shown to improve the outcome of patients affected by a number of inborn errors of metabolism. Recently, MS/MS analytical methods were developed for second-tier tests of newborn screening; new substrates have also been developed to measure the activity of lysosomal enzymes so lysosomal storage diseases can be diagnosed by MS/MS method now.

A faster method to detect norovirus in oysters using probe hybridization to isolate target RNA before RT-PCR.

Human Noroviruses (HuNoVs) are the most frequent cause of outbreaks of acute gastroenteritis following the ingestion of raw or improperly cooked oysters. Although highly sensitive methods to detect HuNoV in oysters using reverse transcriptase-polymerase chain reaction (RT-PCR) are available, rapid methods to process samples for RT-PCR are still needed. The conventional approach is to concentrate the virus first before RNA purification to maximize assay sensitivity, but the procedures used are cumbersome. We developed a new hybridization method that is much faster and more effective compared to existing technology. The procedure includes an initial extraction of total RNA from the digestive diverticula of oysters using TRI Reagent, followed by HuNoV RNA purification using a capture probe and then HuNoV detection by real-time RT-PCR. The detection limit is approximately 100 PCR detection units of HuNoV per sample. Compared to published methods that require an initial virus concentration step before RNA extraction, the new method is much faster to complete. Approximately 3 h are needed to purify HuNoV RNA using the new method compared to at least 8 h using conventional methods. Coupled with real-time RT-PCR, the new method can detect HuNoV in contaminated oysters within 8 h. The effectiveness of the method was demonstrated using live artificially contaminated oysters and wild oysters.

Enhancing PCR amplification of DNA from recalcitrant plant specimens using a trehalose-based additive.

PREMISE OF THE STUDY: PCR amplification of DNA extracted from plants is sometimes difficult due to the presence of inhibitory compounds. An effective method to overcome the inhibitory effect of compounds that contaminate DNA from difficult plant specimens is needed. • METHODS AND RESULTS: The effectiveness of a PCR additive reagent containing trehalose, bovine serum albumin (BSA), and polysorbate-20 (Tween-20) (TBT-PAR) was tested. PCR of DNA extracted from fresh, silica-dried, and herbarium leaf material of species of Achariaceae, Asteraceae, Lacistemataceae, and Samydaceae that failed using standard techniques were successful with the addition of TBT-PAR. • CONCLUSIONS: The addition of TBT-PAR during routine PCR is an effective method to improve amplification of DNA extracted from herbarium specimens or plants that are known to contain PCR inhibitors.

Relationship of human-associated microbial source tracking markers with Enterococci in Gulf of Mexico waters.

Human and ecosystem health can be damaged by fecal contamination of recreational waters. Microbial source tracking (MST) can be used to specifically detect domestic sewage containing human waste, thereby informing both risk assessment and remediation strategies. Previously, an inter-laboratory collaboration developed standardized PCR methods for a bacterial, an archaeal, and a viral indicator of human sewage. Here we present results for two subsequent years of field testing in fresh and salt water by five laboratories across the U.S. Gulf Coast (two in Florida and one each in Mississippi, Louisiana and Texas) using common standard operating procedures (SOPs) developed previously. Culturable enterococci were enumerated by membrane filtration, and PCR was used to detect three MST markers targeting domestic sewage: human-associated Bacteroides (HF183), Methanobrevibacter smithii and human polyomaviruses BK and JC (HPyVs). Detection of sewage markers in surface waters was significantly associated with higher enterococci levels and with exceedance of the recreational water quality standard in four or three regions, respectively. Sewage markers were frequently co-detected in single samples, e.g., M. smithii and HF183 were co-detected in 81% of Louisiana samples, and HPyVs and M. smithii were co-detected in over 40% of southwest Florida and Mississippi samples. This study demonstrates the robustness and inter-laboratory transferability of these three markers for the detection of pollution from domestic sewage in the waters impacting the Gulf of Mexico over a coastal range of over 1000 miles.

A filter-based propidium monoazide technique to distinguish live from membrane-compromised microorganisms using quantitative PCR.

Propidium monoazide (PMA) was used to differentiate live from membrane-compromised bacteria in PCR methods. We have adapted this technique for use on membrane-filtered water samples and determined its efficacy using qPCR. Independent labs at three institutions replicated these findings.