Objective We evaluated the feasibility of the oligonucleotide ligation assay (OLA), a specific, sensitive, and economical ligase-based point mutation assay designed to detect HIV-1 drug–resistance mutations at 12 codons of HIV-1 subtype B pol, for potential use in resource-poor settings. Methods Specimens from HIV-1–infected individuals collected by 7 international laboratories, including subtypes A, B, C, D, F, G, J, and recombinants AE and AG, were tested by the OLA developed for HIV-1 subtype B. Common polymorphisms that interfered with reactivity of the OLA were identified and modified probes designed and evaluated. Results 92.5% (2410) of 2604 codons in specimens from 217 individuals were successfully genotyped by the subtype B OLA. A high rate (range 8.3%–31.2%) of indeterminate results (negative OLA reaction for both mutant and wild type) was observed for 5 codons. Modified probes at reverse transcriptase codons 151 and 184 and protease codon 90 increased the rate of valid OLA to 96.1%. Conclusions The OLA designed for HIV-1 subtype B genotyped most pol codons in non-B subtypes from Asia and Africa but was improved by addition of several modified probes. International laboratories experienced in molecular techniques were able to perform the OLA.

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Background Although many critical care experts and national organizations support open visitation in intensive care units (ICUs), most ICU visiting policies do not allow unrestricted presence of patients' family members. Objective To describe how well the needs of family members were met in an adult neuroscience ICU with a continuous visitation policy and an adjoining private suite for patients' family members. Methods An exploratory, descriptive study design was used to identify the effects of continuous family visitation in the neuroscience ICU on patients' family members and their needs and experiences during their time in the unit. A convenience sample of consenting family members completed a survey of family need items 72 hours after the patient was admitted to the unit. Results The most important needs identified by the 45 family members surveyed were items relating to information about the patient, visiting the patient, being given hope, talking with a doctor each day, and being assured that the best care is being given to the patient. Least important items were related to physical comforts for the family members. The vast majority of family members rated their needs as being met for all of the items in the survey and reported a high level of satisfaction with care. Conclusion In a neuroscience ICU with an open visitation policy and a private suite for patients' family members, family members rated their needs as being met at a high level, unlike in prior studies in units with limitations on family visitation. The rank order of the importance of each need in the survey was similar to rankings in prior studies in a variety of critical care units. (American Journal of Critical Care. 2016;25:118-125) In descriptive research studies [3][4][5][6][7] in the 1970s and 1980s, family members of ICU adult patients consistently identified similar high-priority needs during the time of the critical illness. Some of the highest ranked needs included the need for information about their loved one, to be geographically close to their loved one, and to know that their loved one is receiving the best care possible. About the AuthorsCynthia Horton, Sharon Rance-Ashley, Tera Field, Robbie Patterson, Claudette Johnson, Holly Saunders, Tracy Shelton, and Jessica Miller are staff nurses, Carmen Frobos is a family coordinator, and Mini Jacob is the nurse educator in the neuroscience intensive care unit,

Critical Care 2017, 21(Suppl 1):P349 Introduction Imbalance in cellular energetics has been suggested to be an important mechanism for organ failure in sepsis and septic shock. We hypothesized that such energy imbalance would either be caused by metabolic changes leading to decreased energy production or by increased energy consumption. Thus, we set out to investigate if mitochondrial dysfunction or decreased energy consumption alters cellular metabolism in muscle tissue in experimental sepsis. Methods We submitted anesthetized piglets to sepsis (n = 12) or placebo (n = 4) and monitored them for 3 hours. Plasma lactate and markers of organ failure were measured hourly, as was muscle metabolism by microdialysis. Energy consumption was intervened locally by infusing ouabain through one microdialysis catheter to block major energy expenditure of the cells, by inhibiting the major energy consuming enzyme, N+/K + -ATPase. Similarly, energy production was blocked infusing sodium cyanide (NaCN), in a different region, to block the cytochrome oxidase in muscle tissue mitochondria. Results All animals submitted to sepsis fulfilled sepsis criteria as defined in Sepsis-3, whereas no animals in the placebo group did. Muscle glucose decreased during sepsis independently of N+/K + -ATPase or cytochrome oxidase blockade. Muscle lactate did not increase during sepsis in naïve metabolism. However, during cytochrome oxidase blockade, there was an increase in muscle lactate that was further accentuated during sepsis. Muscle pyruvate did not decrease during sepsis in naïve metabolism. During cytochrome oxidase blockade, there was a decrease in muscle pyruvate, independently of sepsis. Lactate to pyruvate ratio increased during sepsis and was further accentuated during cytochrome oxidase blockade. Muscle glycerol increased during sepsis and decreased slightly without sepsis regardless of N+/K + -ATPase or cytochrome oxidase blocking. There were no significant changes in muscle glutamate or urea during sepsis in absence/presence of N+/K + -ATPase or cytochrome oxidase blockade. ConclusionsThese results indicate increased metabolism of energy substrates in muscle tissue in experimental sepsis. Our results do not indicate presence of energy depletion or mitochondrial dysfunction in muscle and should similar physiologic situation be present in other tissues, other mechanisms of organ failure must be considered. , and long-term follow up has shown increased fracture risk [2]. It is unclear if these changes are a consequence of acute critical illness, or reduced activity afterwards. Bone health assessment during critical illness is challenging, and direct bone strength measurement is not possible. We used a rodent sepsis model to test the hypothesis that critical illness causes early reduction in bone strength and changes in bone architecture. Methods 20 Sprague-Dawley rats (350 ± 15.8g) were anesthetised and randomised to receive cecal ligation and puncture (CLP) (50% cecum length, 18G needle single pass through anterior and posterior wa...

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