L. Desquilbet

Dose-response analyses using restricted cubic spline functions in public health research.

Taking into account a continuous exposure in regression models by using categorization, when non-linear dose-response associations are expected, have been widely criticized. As one alternative, restricted cubic spline (RCS) functions are powerful tools (i) to characterize a dose-response association between a continuous exposure and an outcome, (ii) to visually and/or statistically check the assumption of linearity of the association, and (iii) to minimize residual confounding when adjusting for a continuous exposure. Because their implementation with SAS® software is limited, we developed and present here an SAS macro that (i) creates an RCS function of continuous exposures, (ii) displays graphs showing the dose-response association with 95 per cent confidence interval between one main continuous exposure and an outcome when performing linear, logistic, or Cox models, as well as linear and logistic-generalized estimating equations, and (iii) provides statistical tests for overall and non-linear associations. We illustrate the SAS macro using the third National Health and Nutrition Examination Survey data to investigate adjusted dose-response associations (with different models) between calcium intake and bone mineral density (linear regression), folate intake and hyperhomocysteinemia (logistic regression), and serum high-density lipoprotein cholesterol and cardiovascular mortality (Cox model).

Relationship between a frailty-related phenotype and progressive deterioration of the immune system in HIV-infected men.

Context:Immunological similarities have been noted between HIV-infected individuals and older HIV-negative adults. Immunologic alterations with aging have been noted in frailty in older adults, a clinical syndrome of high risk for mortality and other adverse outcomes. Using a frailty-related phenotype (FRP), we investigated in the Multicenter AIDS Cohort Study whether progressive deterioration of the immune system among HIV-positive individuals independently predicts onset of FRP. Methods:FRP was evaluated semiannually in 1046 HIV-infected men from 1994 to 2005. CD4 T-cell count and plasma viral load were evaluated as predictors of FRP by logistic regression (generalized estimating equations), adjusting for age, ethnicity, educational level, AIDS status, and treatment era [pre-highly active antiretroviral therapy (HAART) (1994-1995) and HAART (1996-1999 and 2000-2005)]. Results:Adjusted prevalences of FRP remained low for CD4 T-cell counts >400 cells per cubic millimeter and increased exponentially and significantly for lower counts. Results were unaffected by treatment era. After 1996, CD4 T-cell count, but not plasma viral load, was independently associated with FRP. Conclusions:CD4 T-cell count predicted the development of a FRP among HIV-infected men, independent of HAART use. This suggests that compromise of the immune system in HIV-infected individuals contributes to the systemic physiologic dysfunction of frailty.

Increase in at-risk sexual behaviour among Hiv-1-infected patients followed in the French Primo cohort

Objective: With the current increase in sexually transmitted infections in industrialized countries, we assessed the characteristics and plasma viral load of HIV-1-infected patients reporting sexual behaviour at risk for HIV transmission (SBR). Design: The study population consisted of 223 patients with primary HIV-1 infection who were enrolled in the French PRIMO cohort between 1996 and 2001 and who had at least 3 months of follow-up. Patients were interviewed on condom use at each visit according to the partner (gender, steady versus casual nature, and HIV serostatus). SBR was defined as unprotected sex with partners of unknown or negative HIV serostatus. Results: Sixty-one SBR were reported by 43 patients. SBR with casual partners increased from 5.1% in 1998 to 21.1% in 2001–2002, after a fall between 1997 and 1998. Reporting of SBR was more frequent among patients with casual partners, those with asymptomatic or briefly (⩽ 15 days) symptomatic primary infection, and those who had developed clinical lipodystrophy or signs of anxiety/depression. Eighty-six per cent of patients reporting SBR had previously initiated highly active antiretroviral therapy (HAART); plasma viral load was above the detection limit (200/500 copies/ml) in 41% of visits reporting an SBR. Viral load was similar in patients reporting SBR and other patients, suggesting that the patients knowledge of his/her response to HAART was not a major determinant of subsequent SBR. Conclusion: Our results confirm the recent increase in unsafe sex observed among HIV-infected individuals in industrialized countries. Levels of viral load of these individuals raise concern about the potential for re-emerging HIV epidemics.

A Frailty-Related Phenotype Before HAART Initiation as an Independent Risk Factor for AIDS or Death After HAART Among HIV-Infected Men

BACKGROUND In the general population, frailty, a late stage of the aging process, predicts mortality. We investigated whether manifesting a previously defined frailty-related phenotype (FRP) before initiating highly active antiretroviral therapy (HAART) affects the likelihood of developing clinical AIDS or mortality after HAART initiation. METHODS Among 596 HIV-infected men in the Multicenter AIDS Cohort Study whose date of HAART initiation was known within ±6 months and who had an assessable FRP status within 3 years before HAART, survival analyses were performed to assess the effect of FRP manifestation on clinical AIDS or death after HAART. RESULTS In men free of AIDS before HAART, AIDS or death after HAART occurred in 13/36 (36%) men who exhibited the FRP before HAART but only in 69/436 (16%) men who did not (hazard ratio = 2.6; 95% confidence interval = 1.4-4.6; p < .01). After adjusting for age, ethnicity, education, nadir CD4+ T-cell count, peak HIV viral load, and hemoglobin in the 3 years before HAART, having the FRP at >25% of visits in the 3 years before HAART significantly predicted AIDS or death (adjusted hazard ratio = 3.8; 95% confidence interval = 1.9-7.9; p < .01). Results were unchanged when the analysis was restricted to the 335 AIDS-free men who were HAART responders, to the 124 men who had AIDS at HAART initiation, or to the subsets of men for whom indices of liver and kidney function could be taken into account. CONCLUSION Having a persistent frailty-like phenotype before HAART initiation predicted a worse prognosis after HAART, independent of known risk factors.

Ground reaction force and kinematic analysis of limb loading on two different beach sand tracks in harness trotters.

REASONS FOR PERFORMING STUDY Although beach training is commonly used in horses, limb loading on beach sand has never been investigated. A dynamometric horseshoe (DHS) is well adapted for this purpose. OBJECTIVES To compare ground reaction force (GRF) and fetlock kinematics measured in harness trotters on 2 tracks of beach sand with different water content. METHODS Two linear sand tracks were compared: firm wet sand (FWS, 19% moisture) vs. deep wet sand (DWS, 13.5% moisture). Four French trotters (550 ± 22 kg) were used. Their right forelimb was equipped with a DHS and skin markers. Each track was tested 3 times at 7 m/s. Each trial was filmed by a high-speed camera (600 Hz); DHS and speed data acquisition was performed at 10 kHz on 10 consecutive strides. All recordings were synchronised. The components Fx (parallel to the hoof solar surface) and Fz (perpendicular) of the GRF were considered. For 3 horses the fetlock angle and forelimb axis-track angle at landing were measured. Statistical differences were tested using the GLM procedure (SAS; P < 0.05). RESULTS Stance duration was increased on DWS compared to FWS. Fzmax and Fxmax (oriented, respectively, downwards and forwards relatively to the solar surface) and the corresponding loading rates, were decreased on DWS and these force peaks occurred later. Fxmin (backwards) was not significantly different between both surfaces; the propulsive phase (Fx negative) was longer and the corresponding impulse higher, on DWS compared to FWS. The forelimb was more oblique to the track at landing and maximal fetlock extension was less and delayed on DWS. CONCLUSIONS This study confirms that trotting on deep sand overall reduces maximal GRF and induces a more progressive limb loading. However, it increases the propulsive effort and likely superficial digital flexor tendon tension at the end of stance, which should be taken into account in beach training.

Rapid CD4+ cell decrease after transient cART initiated during primary HIV infection (ANRS PRIMO and SEROCO cohorts).

Objective:To modelize the rate of CD4+ cell count decline and its determinants after cessation of combination antiretroviral therapy (cART) started during primary HIV infection (PHI) and compare it with never-treated patients. Methods:Kinetics of CD4+ counts were analyzed on the square root scale by using a mixed-effects model in 170 patients who received cART during PHI from the Primary Infection (PRIMO) cohort and 123 never-treated patients from the Seroconverters (SEROCO) cohort. Results:After cART interruption in the PRIMO cohort, the CD4+ cell count fell rapidly during the first 5 months and more slowly thereafter. The timing of treatment initiation had no influence on the rate of CD4+ cell decline. In contrast, a larger increase in CD4+ cell counts during cART was associated with a steeper decline and a larger loss of CD4+ cells after treatment interruption. The mean CD4+ cell loss 3 years postinterruption was 383 cells per microliter. In the SEROCO cohort, the CD4+ T-cell decline was less steep (3-year CD4+ loss 239 cells/μL). As a result, the mean CD4+ cell counts were similar (416 cells/μL) 3 years after cART interruption (PRIMO) or after infection (SEROCO). Conclusions:These data question the benefit of a limited course of cART even when initiated within 3 months after PHI diagnosis.

Effect of endocervical inflammation on days to conception in dairy cows

In contrast to endometritis, now diagnosed by cytological examination, the effect of endocervical inflammation on reproductive performance has been inadequately investigated. In this study, endocervical and endometrial cytological specimens were collected from 168 Holstein cows between 21 and 60 days in milk (DIM) to investigate the prevalence of endocervical inflammation and effect on days to conception. Statistical analyses were stratified based on DIM at examination (<35 vs. ≥35 DIM). Endocervical inflammation with ≥5% neutrophils before 35 DIM (disregarding the level of endometrial inflammation) was associated with decreased hazard of pregnancy within 300 DIM (adjusted hazard ratio, 0.4; 95% confidence interval: 0.3-0.8). A decrease in hazard of pregnancy was observed when >6% neutrophils were counted in endometrial smears (adjusted hazard ratio, 0.4; 95% confidence interval: 0.2-0.7). The study suggested an additive effect of combined endocervical and endometrial inflammation on the hazard of pregnancy within 300 DIM. Using the thresholds of 5% neutrophils for the cervix and 6% neutrophils for the uterus, 11% of the cows examined before 35 DIM presented cervicitis only, 13% were affected by endometritis only, and 32% suffered from both endometrial and endocervical inflammation. The presence (absence) of cervicitis was not indicative of the presence (absence) of endometritis. This study showed that in addition to uterine inflammation, endocervical inflammation in early lactation affects conception. Thus, the global evaluation of genital tract health may be more beneficial for reproductive performance than that of endometrial inflammation.

Disruption of the Bcchs3a Chitin Synthase Gene in Botrytis cinerea Is Responsible for Altered Adhesion and Overstimulation of Host Plant Immunity

The fungal cell wall is a dynamic structure that protects the cell from different environmental stresses suggesting that wall synthesizing enzymes are of great importance for fungal virulence. Previously, we reported the isolation and characterization of a mutant in class III chitin synthase, Bcchs3a, in the phytopathogenic fungus Botrytis cinerea. We demonstrated that virulence of this mutant is severely impaired. Here, we describe the virulence phenotype of the cell-wall mutant Bcchs3a on the model plant Arabidopsis thaliana and analyze its virulence properties, using a variety of A. thaliana mutants. We found that mutant Bcchs3a is virulent on pad2 and pad3 mutant leaves defective in camalexin. Mutant Bcchs3a was not more susceptible towards camalexin than the wild-type strain but induced phytoalexin accumulation at the infection site on Col-0 plants. Moreover, this increase in camalexin was correlated with overexpression of the PAD3 gene observed as early as 18 h postinoculation. The infection process of the mutant mycelium was always delayed by 48 h, even on pad3 plants, probably because of lack of mycelium adhesion. No loss in virulence was found when Bcchs3a conidia were used as the inoculum source. Collectively, these data led us to assign a critical role to the BcCHS3a chitin synthase isoform, both in fungal virulence and plant defense response.

Predictive markers of clinical outcome in the GRMD dog model of Duchenne muscular dystrophy

In the translational process of developing innovative therapies for DMD (Duchenne muscular dystrophy), the last preclinical validation step is often carried out in the most relevant animal model of this human disease, namely the GRMD (Golden Retriever muscular dystrophy) dog. The disease in GRMD dogs mimics human DMD in many aspects, including the inter-individual heterogeneity. This last point can be seen as a drawback for an animal model but is inherently related to the disease in GRMD dogs closely resembling that of individuals with DMD. In order to improve the management of this inter-individual heterogeneity, we have screened a combination of biomarkers in sixty-one 2-month-old GRMD dogs at the onset of the disease and a posteriori we addressed their predictive value on the severity of the disease. Three non-invasive biomarkers obtained at early stages of the disease were found to be highly predictive for the loss of ambulation before 6 months of age. An elevation in the number of circulating CD4+CD49dhi T cells and a decreased stride frequency resulting in a reduced spontaneous speed were found to be strongly associated with the severe clinical form of the disease. These factors can be used as predictive tests to screen dogs to separate them into groups with slow or fast disease progression before their inclusion into a therapeutic preclinical trial, and therefore improve the reliability and translational value of the trials carried out on this invaluable large animal model. These same biomarkers have also been described to be predictive for the time to loss of ambulation in boys with DMD, strengthening the relevance of GRMD dogs as preclinical models of this devastating muscle disease.

A restricted cubic spline approach to assess the association between high fat fish intake and red blood cell EPA + DHA content

BACKGROUND AND AIMS Fish, especially fatty fish, are the main contributor to eicosapentaenoic (EPA) and docosahexaenoic (DHA) intake. EPA and DHA concentrations in red blood cells (RBC) has been proposed as a cardiovascular risk factor, with <4% and >8% associated with the lowest and greatest protection, respectively. The relationship between high fat fish (HFF) intake and RBC EPA + DHA content has been little investigated on a wide range of fish intake, and may be non-linear. We aimed to study the shape of this relationship among high seafood consumers. METHODS AND RESULTS Seafood consumption records and blood were collected from 384 French heavy seafood consumers and EPA and DHA were measured in RBC. A multivariate linear regression was performed using restricted cubic splines to consider potential non-linear associations. Thirty-six percent of subjects had an RBC EPA + DHA content lower than 4% and only 5% exceeded 8%. HFF consumption was significantly associated with RBC EPA + DHA content (P [overall association] = 0.021) adjusted for sex, tobacco status, study area, socioeconomic status, age, alcohol, other seafood, meat, and meat product intakes. This relationship was non-linear: for intakes higher than 200 g/wk, EPA + DHA content tended to stagnate. Tobacco status and fish contaminants were negatively associated with RBC EPA + DHA content. CONCLUSION Because of the saturation for high intakes, and accounting for the concern with exposure to trace element contaminants, intake not exceeding 200 g should be considered.