Combined germline and somatic human FADD mutations cause autoimmune lymphoproliferative syndrome.

BACKGROUND: The autoimmune lymphoproliferative syndrome (ALPS) is a noninfectious and nonmalignant lymphoproliferative disease frequently associated with autoimmune cytopenia resulting from defective FAS signaling. We previously described germline monoallelic FAS (TNFRSF6) haploinsufficient mutations associated with somatic events, such as loss of heterozygosity on the second allele of FAS, as a cause of ALPS-FAS. These somatic events were identified by sequencing FAS in DNA from double-negative (DN) T cells, the pathognomonic T-cell subset in ALPS, in which the somatic events accumulated. OBJECTIVE: We sought to identify whether a somatic event affecting the FAS-associated death domain (FADD) gene could be related to the disease onset in 4 unrelated patients with ALPS carrying a germline monoallelic mutation of the FADD protein inherited from a healthy parent. METHODS: We sequenced FADD and performed array-based comparative genomic hybridization using DNA from sorted CD4+ or DN T cells. RESULTS: We found homozygous FADD mutations in the DN T cells from all 4 patients, which resulted from uniparental disomy. FADD deficiency caused by germline heterozygous FADD mutations associated with a somatic loss of heterozygosity was a phenocopy of ALPS-FAS without the more complex symptoms reported in patients with germline biallelic FADD mutations. CONCLUSIONS: The association of germline and somatic events affecting the FADD gene is a new genetic cause of ALPS.

Evidence of inter- and intra-keloid heterogeneity through analysis of dermal fibroblasts: A new insight in deciphering keloid physiopathology.

Keloid scars are hypertrophic and proliferating pathological scars extending beyond the initial lesion and without tendency to regression. Usually, keloids are considered and treated as a single entity but clinical observations suggest heterogeneity in keloid morphologies with distinction of superficial/extensive and nodular entities. Within a keloid, heterogeneity could also be detected between superficial and deep dermis or centre and periphery. Focusing on fibroblasts as main actors of keloid formation, we aimed at evaluating intra- and inter-keloid fibroblast heterogeneity by analysing their gene expression and functional capacities (proliferation, migration, traction forces), in order to improve our understanding of keloid pathogenesis. Fibroblasts were obtained from centre, periphery, papillary and reticular dermis from extensive or nodular keloids and were compared to control fibroblasts from healthy skin. Transcriptional profiling of fibroblasts identified a total of 834 differentially expressed genes between nodular and extensive keloids. Quantification of ECM-associated gene expression by RT-qPCR brought evidence that central reticular fibroblasts of nodular keloids are the population which synthesize higher levels of mature collagens, TGFß, HIF1α and αSMA as compared to control skin, suggesting that this central deep region is the nucleus of ECM production with a centrifuge extension in keloids. Although no significant variations were found for basal proliferation, migration of peripheral fibroblasts from extensive keloids was higher than that of central ones and from nodular cells. Moreover, these peripheral fibroblasts from extensive keloids exhibited higher traction forces than central cells, control fibroblasts and nodular ones. Altogether, studying fibroblast features demonstrate keloid heterogeneity, leading to a better understanding of keloid pathophysiology and treatment adaptation.

Multilayer spectrum of intratumoral heterogeneity in basal bladder cancer.

Basal/squamous (Ba/Sq) subtype represents an intrinsic and robust group in the consensus molecular classification of muscle-invasive bladder cancer (MIBC), with poor outcome and controversial chemosensitivity. We aimed to investigate the spectrum of intratumor heterogeneity (ITH) in the Ba/Sq subtype. First, we validated a 29-gene NanoString CodeSet to predict the Ba/Sq subtype for FFPE samples. We identified heterogeneous Ba/Sq tumors in a series of 331 MIBC FFPE samples using dual GATA3/KRT5/6 immunohistochemistry (IHC). Heterogeneous regions with distinct immunostaining patterns were studied separately for gene expression using the 29-gene CodeSet, for mutations by targeted next-generation sequencing, and for copy number alteration (CNA) by microarray hybridization. Among 83 Ba/Sq tumors identified by GATA3/KRT5/6 dual staining, 19 tumors showed heterogeneity at the IHC level. In one third of the 19 cases, regions from the same tumor were classified in different distinct molecular subtypes. The mutational and CNA profiles confirmed the same clonal origin for IHC heterogeneous regions with possible subclonal evolution. Overall, two patterns of intratumoral heterogeneity (ITH) were observed in Ba/Sq tumors: low ITH (regions with distinct immunostaining, but common molecular subtype and shared CNA) or high ITH (regions with distinct immunostaining, molecular subtype, and CNA). These results showed multilayer heterogeneity in Ba/Sq MIBC. In view of personalized medicine, this heterogeneity adds complexity and should be taken into account for sampling procedures used for diagnosis and treatment choice. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Prediction and Comparison of Fentanyl Infusion Pharmacokinetics in Obese and Nonobese Children.

OBJECTIVES: To compare fentanyl infusion pharmacokinetic variables in obese children and nonobese children. DESIGN: A pharmacokinetic simulation study. SETTING: We used a semi-physiologically based pharmacokinetic model to generate fentanyl pharmacokinetic variables. SUBJECTS: Simulations of pharmacokinetic variables were based on historical inpatient demographic data in less than 18-year-olds. INTERVENTIONS: Obese children were defined as children less than 2 years with weight-for-length greater than or equal to 97.7th percentile or body mass index-for-age greater than or equal to 95th percentile for greater than or equal to 2-17-year-olds. MEASUREMENTS AND MAIN RESULTS: Overall, 4,376 patients were included, with 807 (18.4%) classified as obese children. The majority (52.9%) were male, with a median age of 8.1 years (interquartile range, 4.3-13.0 yr). The differences in total clearance (CLS), volume of distribution at steady-state values, weight-normalized CLS, and weight-normalized volume of distribution at steady state were assessed in obese children and nonobese children. Multivariable analyses indicated that obesity was significantly associated with a higher CLS in obese children greater than 6-year-olds (p < 0.0375). However, there was an 11-30% decrease in weight-normalized CLS in obese children versus nonobese children in all age groups (p < 0.05). Both volume of distribution at steady state and weight-normalized volume of distribution at steady state increased significantly in obese children compared with nonobese children (p < 0.05). Fentanyl plasma concentration-time profiles of obese children and nonobese children pairs (ages 4, 9, and 15) receiving 1 µg/kg/hr using total body weight were also compared. Steady-state concentrations of the obese children using similar weight-based dosing increased by 25%, 77%, and 44% in comparison to nonobese children 4-, 9-, and 15-year-olds, respectively. Time to steady state and elimination half-lives were two- to four-fold longer in obese children. An additional simulation was conducted for 15-year-old obese children and nonobese children using a fixed dose of 50 µg/hr and it provided similar pharmacokinetic profiles. CONCLUSIONS: CLS may increase less than proportional to weight in obese children greater than 6-year-olds, while volume of distribution at steady state increases more than proportional to weight in all obese children compared with nonobese children. Weight-based dosing in obese children may cause an increase in steady-state concentration while prolonging the time to steady state. Exploring alternative dosing strategies for obese children is warranted.

Self-Esteem and HIV Infection in Morocco: Associated Factors Among People Living with HIV-Results from a Community-Based Study.

People living with HIV (PLHIV) face specific issues regarding mental quality of life (QoL), in particular self-esteem. The objective of this study was to measure self-esteem and to identify associated factors among PLHIV in Morocco. A 125-item questionnaire was administered to 300 PLHIV. The dependent variable was adapted from Rosenberg's self-esteem scale (range 0-4). A weighted multiple linear regression was performed. The mean level of self-esteem was 2.4 ± 1.0. The factors independently associated with self-esteem were: feeling of loneliness (p = 0.001), perceived seriousness of infection (p = 0.006), thinking serostatus disclosure was a mistake (p = 0.007), thinking HIV infection will last for life (p = 0.008), sexual orientation (p = 0.050), satisfaction with sexual life (p = 0.019) and perceived treatment efficacy (p = 0.009). These results underline the need for evidence-based interventions (e.g. anti-discrimination measures, interventions to prevent social isolation of PLHIV, support in the serostatus disclosure process), in order to improve the social environment and eventually improve their self-esteem and QoL.

HIV seropositivity and sexuality: cessation of sexual relations among men and women living with HIV in five countries.

The sexuality of people living with HIV (PLHIV) is a key issue in the fight against HIV, as it influences both the dynamic of the epidemic and the quality of life of PLHIV. The present study examined the factors associated with cessation of sexual relations after HIV diagnosis among men and women in five countries: Mali, Morocco, Democratic Republic of the Congo, Romania and Ecuador. A community-based cross-sectional study was implemented by a mixed consortium [researchers/community-based organizations (CBO)]. Trained CBO members interviewed 1500 PLHIV in contact with CBOs using a 125-item questionnaire. A weighted multivariate logistic regression and a separate gender analysis were performed. Among the 1413 participants, 471 (33%) declared that they stopped having sexual relations after their HIV diagnosis, including 318 women (42%) and 153 men (23%) (p < .001). Concerning women, variables associated with the cessation of sexual relations in the final multivariate model were mainly related with relational factors and the possibility of getting social support (e.g., needing help to disclose HIV serostatus, feeling lonely every day, not finding support in CBOs, not being in a couple). Men's sexual activity was more associated with their representations and their perception of the infection (e.g., thinking they will have their HIV infection for the rest of their life, perceiving the HIV infection as a mystery, perceiving the infection as serious). Furthermore, the following variables were associated with both men and women sexual behaviours: being older, having suffered from serious social consequences after serostatus disclosure and not being able to regularly discuss about HIV with their steady partner. Results suggested clear differences between men and women regarding cessation of sexual relations and highlighted the importance of implementing gender-based tailored interventions that promote safe and satisfying sexuality, as it is known to have a positive impact on the overall well-being of PLHIV.

FACTORS ASSOCIATED WITH HIV VOLUNTARY DISCLOSURE TO ONE'S STEADY SEXUAL PARTNER IN MALI: RESULTS FROM A COMMUNITY-BASED STUDY.

Despite the widespread dissemination of HIV information through public awareness campaigns in Mali, disclosing seropositivity to one's steady sexual partner (SSP) remains difficult for people living with HIV (PLHIV). Disclosure is a public health concern with serious implications and is also strongly linked to the quality of life of PLHIV. This study aimed to analyse factors associated with voluntary HIV disclosure to one's SSP, using a community-based cross-sectional study on 300 adult PLHIV in contact with a Malian community-based organization working in the field of AIDS response. A 125-item questionnaire was administered by trained personnel to study participants between May and October 2011. Analysis was restricted to the 219 participants who both reported having a SSP and answered to the question on disclosure to their SSP. A weighted multivariate logistic regression was used to determine variables independently associated with disclosure. In total, 161 participants (73%) reported HIV disclosure to their SSP. Having children (odds ratio [95% confidence interval]: 4.52 [1.84-11.12]), being accompanied to the survey site (3.66 [1.00-13.33]), knowing others who had publicly declared their seropositivity (3.12 [1.59-6.12]), having higher self-esteem (1.55 [1.09-2.19]) and using means other than anti-retroviral treatment to treat HIV (0.33 [0.11-1.00]) were independently associated with disclosure. This study identified several factors that should be considered for the design of interventions aimed at facilitating disclosure if/when desired in this cultural context.

"Was it a mistake to tell others that you are infected with HIV?": factors associated with regret following HIV disclosure among people living with HIV in five countries (Mali, Morocco, Democratic Republic of the Congo, Ecuador and Romania). Results from a community-based research.

This study examined regret following HIV serostatus disclosure and associated factors in under-investigated contexts (Mali, Morocco, Democratic Republic of the Congo, Ecuador and Romania). A community-based cross-sectional study was implemented by a mixed consortium [researchers/community-based organizations (CBO)]. Trained CBO members interviewed 1,500 PLHIV in contact with CBOs using a 125-item questionnaire. A weighted multivariate logistic regression was performed. Among the 1,212 participants included in the analysis, 290 (23.9 %) declared that disclosure was a mistake. Female gender, percentage of PLHIV's network knowing about one's seropositivity from a third party, having suffered rejection after disclosure, having suffered HIV-based discrimination at work, perceived seriousness of infection score, daily loneliness, property index and self-esteem score were independently associated with regret. Discrimination, as well as individual characteristics and skills may affect the disclosure experience. Interventions aiming at improving PLHIV skills and reducing their social isolation may facilitate the disclosure process and avoid negative consequences.

[Factors associated with contacting HIV/AIDS associations in Ecuador: results of a community study]. / Factores asociados a establecer contacto con asociaciones de lucha contra el VIH/sida en Ecuador: resultados de un estudio comunitario.

OBJECTIVE: To describe the profile of people living with HIV/AIDS (PLHA) who contact HIV/AIDS associations in Ecuador and to identify the factors related to that contact. METHODS: In 2011, a cross-sectional community study was conducted in two hospitals in Guayaquil. Based on a 125-question survey administered to 300 adult PLHA, a weighted multivariate regression analysis was performed to identify the factors related to contact with an association RESULTS: Of the 300 participants, 34 (11.3%) were in contact with an association. Being over the age of 35, scoring high on the self-efficacy scale, having suffered serious social consequences after disclosing their HIV status, being able to talk to friends about living with HIV, expressing the need to talk about living with HIV with a health professional, and scoring low on the index of the HIV status disclosure control effort were related to that contact. CONCLUSIONS: The characteristics of the PLHA in contact with an association were: being over the age of 35, having suffered serious social consequences following disclosure of seropositivity, and feeling that their psychosocial needs were not being met by the health system in terms of the services provided. These people more easily managed their HIV status in their social milieu and displayed a greater ability to talk to friends about their seropositivity. This information is useful for community actors to maintain and support mobilization on HIV in Ecuador.

Satisfaction with sexual life in people living with HIV in Romania, together with associated individual and social factors.

The present study aimed to identify social and individual factors associated with satisfaction with sexual life (SSL) in people living with HIV (PLHIV) in contact with a community-based organisation in Romania. A standardised questionnaire was administered (N = 300) in a community-based research study. Multivariate analysis (using a weighted logistic regression restricted to the 291 respondents who answered a question about SSL) was used to determine factors associated with SSL. Sixty-eight per cent of the participants declared that they were satisfied with their sexual life. The following individual factors were associated with SSL: being over 36 years old (Odds Ratio [95% CI]: 0.27 [0.13-0.55]), having ceased sexual intercourse because of HIV (0.33 [0.14-0.76]), not knowing how infection had occurred (0.29 [0.15-0.59]), being officially registered with a level of disability lower than "severe" (0.47 [0.23-0.98]) and having a higher self-efficacy score (1.36 [1.14-1.61]). Living in a couple (7.60 [3.69-15.66]), knowing at least one HIV-infected person who had publicly disclosed his/her seropositivity (2.23 [1.03-4.84]), and having a higher social exclusion score (0.91 [0.82-1]) were social factors associated with SSL. The results suggest that HIV service providers must be sensitised to the necessity of systematically including the topic of PLHIV SSL in field interventions. Self-empowerment, positive examples of public disclosure, promoting the benefits of living in a couple, and supporting social integration can all improve the well-being of PLHIV, including their SSL.

Factors associated with HIV status disclosure to one's steady sexual partner in PLHIV in Morocco.

The objective of the present study was to determine the factors independently associated with disclosure of seropositivity to one's steady sexual partner in people living with HIV (PLHIV) who are recipients of services provided by Association de Lutte Contre le Sida, a Moroccan community-based organization (CBO) working on AIDS response. Between May and October 2011, 300 PLHIV were interviewed about their sociodemographic and economic characteristics, their sexual life and disclosure of their serostatus to their friends, family and to their steady sexual partner. A weighted logistic regression was used to study factors associated with serostatus disclosure to one's steady sexual partner. We restricted the analysis to people who declared they had a steady sexual partner (n = 124). Median age was 36 years old, 56 % were men and 62 % declared that they had disclosed their serostatus to their steady sexual partner. The following factors were independently associated with disclosure: living with one's steady sexual partner [OR 95 % CI: 9.85 (2.86-33.98)], having a higher living-standard index [2.06 (1.14-3.72)], regularly discussing HIV with friends [6.54 (1.07-39.77)] and CBO members [4.44 (1.27-15.53)], and having a higher social exclusion score [1.24 (1.07-1.44)]. Unemployment (as opposed to being a housewife) was negatively associated with disclosure [0.12 (0.02-0.87)]. Despite the potential positive effects for the prevention of HIV transmission and for adherence to HIV treatment, many PLHIV had not disclosed their serostatus to their steady sexual partner. Some factors shown here to be significantly associated with such disclosure will help in the development of future support interventions.

[Facilitating access to care for most-at-risk populations : the Bamako night sexual health clinic experience (Mali)]. / Faciliter l'accès aux soins des populations les plus exposées : l'expérience de la clinique nocturne de santé sexuelle de Bamako au Mali.

The estimated prevalence of HIV in Mali is 1.3 % of the general population. The epidemic is concentrated in certain groups, particularly men who have sex with men (MSM) and sex workers (SW). Access to care is limited for these populations, notably because of structural obstacles (e.g. marked social rejection ; health care services poorly adapted to the real needs of these people). Innovative strategies must be envisaged to ensure access to care services and retention in care for these populations. As part of a health promotion process, ARCAD-SIDA, a Malian NGO involved in the fight against AIDS since 1995, set up a night sexual health clinic in 2010 as part of a strategy to more adequately respond to the health needs of these populations. This clinic adapts health service timetables to match the lifestyles of the targeted populations, brings services in closer physical proximity to the places in which these populations live, proposes patient-tailored consultations, works to improve the patients' psychosocial skills, and promotes community-based peer mobilization. In an environment which is generally hostile to MSM and SW, ARCAD-SIDA also works in advocacy, targeting political decision-makers, defense forces and journalists. The NGO has also played a key role in ensuring that these populations are taken into account in the national strategy for the fight against HIV. Since opening in 2010, the clinic has helped reach a large number of MSM and SW and has improved retention in care. This innovative strategy has also enabled the NGO to improve its professional practices in terms of an individual-based approach to prevention. Interventions that are better adapted to the needs and environment of the populations for whom they are intented to have a positive effect on access to and use of healthcare services.

Bilateral pulmonary artery banding in higher risk neonates with hypoplastic left heart syndrome.

Objectives: Limited data on performing bilateral pulmonary artery banding (BPAB) before stage 1 Norwood procedure suggest that some patients may benefit through the postponement of the major cardiopulmonary bypass procedure. The objective of this study was to evaluate the effectiveness of BPAB in the surgical management of high-risk patients with hypoplastic left heart syndrome (HLHS). Methods: A retrospective review of all high-risk neonates with HLHS who underwent BPAB at our institution was performed. No patients, including those with intact or highly restrictive atrial septum (IAS), were excluded. Results: Between October 2015 and April 2021, 49 neonates with HLHS (including 6 with IAS) underwent BPAB, 40 of whom progressed to the Norwood procedure. Risk factors for not progressing to the Norwood procedure after BPAP include low birth weight (P = .043), the presence of multiple extracardiac anomalies (P = .005), and the presence of genetic disorders (P = .028). Operative mortality was 7.5% (3/40). IAS was associated with operative mortality (P = .022). Conclusions: The strategy of BPAB prestage 1 Norwood procedure was successful in identifying at-risk patients and improving Norwood survival. Although not all patients will need this hybrid approach, a significant number can be expected to benefit from this tactic. These results support the need for a substantial hybrid strategy, in addition to a primary stage 1 Norwood surgical strategy, in the management of HLHS.

Patient Derived Xenografts (PDX) Models as an Avatar to Assess Personalized Therapy Options in Uveal Melanoma: A Feasibility Study.

Uveal melanoma is the most common primary intraocular malignancy in adults. Up to 50% of UM patients develop metastatic disease, usually in the liver. When metastatic, the prognosis is poor, and few treatment options exist. Here, we investigated the feasibility of establishing patient-derived xenografts (PDXs) from a patient's tumor in order to screen for therapies that the patient could benefit from. Samples obtained from 29 primary tumors and liver metastases of uveal melanoma were grafted into SCID mice. PDX models were successfully established for 35% of primary patient tumors and 67% of liver metastases. The tumor take rate was proportional to the risk of metastases. PDXs showed the same morphology, the same GNAQ/11, BAP1, and SF3B1 mutations, and the same chromosome 3 and 8q status as the corresponding patient samples. Six PDX models were challenged with two compounds for 4 weeks. We show that, for 31% of patients with high or intermediate risk of metastasis, the timing to obtain efficacy results on PDX models derived from their primary tumors was compatible with the selection of the therapy to treat the patient after relapse. PDXs could thus be a valid tool ("avatar") to select the best personalized therapy for one third of patients that are most at risk of relapse.

Multi-omics comparison of malignant and normal uveal melanocytes reveals molecular features of uveal melanoma.

Uveal melanoma (UM) is a rare cancer resulting from the transformation of melanocytes in the uveal tract. Integrative analysis has identified four molecular and clinical subsets of UM. To improve our molecular understanding of UM, we performed extensive multi-omics characterization comparing two aggressive UM patient-derived xenograft models with normal choroidal melanocytes, including DNA optical mapping, specific histone modifications, and DNA topology analysis using Hi-C. Our gene expression and cytogenetic analyses suggest that genomic instability is a hallmark of UM. We also identified a recurrent deletion in the BAP1 promoter resulting in loss of expression and associated with high risk of metastases in UM patients. Hi-C revealed chromatin topology changes associated with the upregulation of PRAME, an independent prognostic biomarker in UM, and a potential therapeutic target. Our findings illustrate how multi-omics approaches can improve our understanding of tumorigenesis and reveal two distinct mechanisms of gene expression dysregulation in UM.

Proximity to HIV is associated with a high rate of HIV testing among men who have sex with men living in Douala, Cameroon.

In low- and middle-income countries, men who have sex with men (MSM) are 19 times more likely to be HIV positive compared with background populations. Criminalisation and social rejection of homosexuality in most sub-Saharan African countries reinforce stigma and exclude MSM from prevention activities, including HIV testing. This paper's purpose is to identify factors associated with never having been HIV tested (NHT), among a sample of Cameroonian MSM. In 2008, a community-based study was conducted in Douala, the economic capital city of Cameroon, by a local NGO Alternatives-Cameroun, recruiting participants through the snowball technique and administering a questionnaire during face-to-face interviews. Proximity to HIV was investigated according to the following criteria: knowing at least one person living with HIV and having been exposed to HIV prevention interventions. NHT was defined as reporting to have never been HIV tested. A logistic regression was used to identify factors associated with NHT. Among the 165 MSM of our study group who reported that they were not HIV positive, 19% reported NHT. Factors independently associated with NHT were as follows: being younger, being Muslim, not having a steady male partner, not knowing any person living with HIV and never having been exposed to HIV prevention interventions. In this MSM population, a small proportion reported that they had never been HIV tested and among these, the percentage was higher among individuals not in proximity to HIV. Despite the hostile context of sub-Saharan African countries towards MSM, local and national HIV testing campaigns to date may have played a substantial role in raising HIV awareness in the MSM population living in Douala, and peer-based counselling may have educated those in contact with Alternatives-Cameroun regarding the positive value of HIV testing. This result is a further argument for continuing community-based prevention and extending it to difficult-to-reach MSM.

Analysis of fentanyl pharmacokinetics, and its sedative effects and tolerance in critically ill children.

INTRODUCTION: Fentanyl pharmacokinetic and pharmacodynamic data are limited in mechanically ventilated children. This study aimed to assess the fentanyl pharmacokinetics (PK), the sedation outcome, and the development of tolerance in children receiving fentanyl continuous infusion. METHODS: This study included children admitted to the pediatric or cardiovascular intensive care unit between January 1 and October 31, 2016, who were >30 days to <18 years of age, receiving ventilatory support via endotracheal tube or tracheostomy, and receiving a fentanyl infusion. Population PK analysis was performed using a nonlinear mixed-effects model. The relationship between initial sedation outcome using State Behavioral Scale (SBS) and fentanyl exposure was assessed, and the observations consistent with tolerance were described. RESULTS: Seventeen children, with a median age of 0.83 years (range: 0.1-12) and weight of 8.7 kg (range: 3.4-52), were included. The fentanyl PK was adequately described by a weight-based allometry model with the power of 0.75 for clearance (CL=89.8 L/hr/70 kg) and distributional CL, and 1 for volumes of distribution. In infants <6.6 months, age was an additional factor for CL (31.4 L/h/70 kg) to account for age-related maturation. Seven of twelve nonparalyzed patients achieved goal sedation, defined as >80% of SBS scores ≤0 per 24 h, on the first day of fentanyl infusion with a median plasma concentration of 1.29 ng/ml (interquartile range: 0.78-2.05). Eight of the nine tolerant patients developed tolerance within a day of reaching goal sedation. CONCLUSION: Different weight-based fentanyl dosing rates may be required for infants and children of different ages to achieve similar plasma concentrations. Using SBS scores may guide the dosing titration of fentanyl that resulted in plasma concentrations within the therapeutic range of 1-3 ng/ml. For those who developed tolerance to fentanyl and/or a sedative, it was noted one day after goal sedation was achieved.

Digital Multiplexed Gene Expression Analysis of mRNA and miRNA from Routinely Processed and Stained Cytological Smears: A Proof-of-Principle Study.

OBJECTIVE: Although transcriptomic assessments of small samples using high-throughput techniques are usually performed on fresh or frozen tissues, there is a growing demand for those performed on stained cellular specimens already used for diagnostic purposes. STUDY DESIGN: The possibility of detecting mRNAs and microRNAs (miRNAs) from routinely processed cytological samples using nCounter® technology was explored. Fresh samples from pleural and peritoneal effusions were analyzed using 2 parallel methods: samples were smeared and routinely stained using the May-Grünwald-Giemsa or Diff-Quik® method and mounted using conventional methods, and they were also studied following a snap freezing method, in which samples were maintained at -80°C until use. mRNAs and miRNAs were assessed and compared after total RNA extraction from both routinely processed samples and their matched frozen controls. RESULTS: A good concordance was found between the gene expression measured in routinely processed samples and their matched frozen controls for the majority of mRNAs and miRNAs tested. However, the standard deviation of low-expressed miRNA was high. CONCLUSIONS: Although nCounter® technology is a robust method to measure and characterize both mRNAs and miRNAs from routinely processed cytological samples, caution is recommended for the interpretation of low-expressed miRNA.

Monogeneans from Epinephelus chlorostigma (Val.) (Perciformes: Serranidae) off New Caledonia, with the description of three new species of diplectanids.

Gill monogeneans from the brownspotted grouper Epinephelus chlorostigma (Val.) collected in deep water off the coral barrier reef of New Caledonia, South Pacific, comprise seven species. These include the ancyrocephalid Haliotrema sp., the capsalid Allobenedenia cf. epinepheli Yamaguti, 1968, and five diplectanids, namely Pseudorhabdosynochus epinepheli (Yamaguti, 1938), reported in a previous paper, P. cyanopodus Sigura & Justine, 2008 and P. podocyanus Sigura & Justine, 2008, two species originally described from E. cyanopodus Richardson, P. stigmosus n. sp., P. exoticoides n. sp. and Diplectanum femineum n. sp. P. stigmosus is characterised by a sclerotised vagina with a straight primary canal, large ovoid primary chamber and spherical secondary chamber. P. exoticoides is a highly aberrant species, with a thick-walled male quadriloculate organ and a discoid sclerotised vagina with an exceptional structure. Interestingly, P. exoticoides resembles P. exoticus Sigura & Justine, 2008, a species from E. cyanopodus, and P. stigmosus resembles P. cyanopodus and P. podocyanus, also both from E. cyanopodus, suggesting close relationships between the diplectanid faunae of these two fish species. D. femineum belongs to a group of diplectanids, provisionally classified as 'Diplectanum' Diesing, 1858, which all share a small funnel-shaped male copulatory organ. In contrast to other members of this group which have no sclerotised vagina, D. femineum has a sclerotised vagina with the same organisation as those of species of Pseudorhabdosynochus Yamaguti, 1958. This suggests that the species of 'Diplectanum' from groupers are closer to Pseudorhabdosynochus than suggested by the structure of the male organs.

Empowering Malian women living with HIV regarding serostatus disclosure management: Short-term effects of a community-based intervention.

OBJECTIVE: The objective of this study was to assess the short-term effects of Gundo-So-a program aimed at empowering Malian women living with HIV (WLHIV) regarding serostatus disclosure management. METHODS: A pre-experimental study with two measures (one week before and four weeks after Gundo-So) was carried out. A 35-item questionnaire was administered to a convenience sample of 210 WLHIV. Six outcomes were considered: ability to decide whether or not to disclose HIV status, self-efficacy to keep HIV status a secret, self-efficacy to disclose HIV status, feeling crushed by the weight of secrecy, perceived physical health, and perceived psychological health. For each outcome, temporal changes associated with the intervention were assessed using linear regressions with random intercepts. RESULTS: Statistically significant change was observed for all six outcomes between the pre- and post-intervention measures. Furthermore, several variables were associated with the baseline levels of the outcomes and the intervention effect. CONCLUSION: The results suggest that Gundo-So empowers Malian WLHIV with regard to serostatus disclosure management, thus improving their perceived physical and psychological health. PRACTICAL IMPLICATIONS: These results highlight the need for programs to empower WLHIV regarding serostatus disclosure, so that WLHIV can make free and informed decisions regarding serostatus disclosure.