[Social medicine: places, collaborations, human-centered practices]. / Médecine sociale : des lieux, des collaborations, des pratiques centrées sur l'humain.

People in complex social situations are often confronted with multiple health problems related to their living conditions, pathologies, addictions and other co-morbidities. They need multi-professional support, while respecting the ethics of care, and in coordination with social partners. Various dedicated services exist, in which nurses are very present.

In vitro, in cellulo and structural characterizations of the interaction between the integrase of Porcine Endogenous Retrovirus A/C and proteins of the BET family.

Retroviral integrase (IN) proteins catalyze the permanent integration of the viral genome into host DNA. They can productively recruit cellular proteins, and the human Bromodomain and Extra-Terminal domain (hBET) proteins have been shown to be co-factors for integration of gamma-retroviruses such as Murine Leukemia Virus (MLV) into human cells. By using two-hybrid, co-immunoprecipitation and in vitro interaction assays, we showed that IN of the gamma- Porcine Endogenous Retrovirus-A/C (PERV IN) interacts through its C-terminal domain (CTD) with hBET proteins. We observed that PERV IN interacts with the BRD2, BRD3 and BRD4 proteins in vitro and that the BRD2 protein specifically binds and co-localizes with PERV IN protein in the nucleus of cells. We further mapped the interaction sites to the conserved Extra-Terminal (ET) domain of the hBET proteins and to several amino acids of the of the C-terminal tail of the PERV IN CTD. Finally, we determined the first experimental structure of an IN CTD - BET ET complex from small-angle X-ray scattering data (SAXS). We showed that the two factors assemble as two distinct modules linked by a short loop which confers partial flexibility. The SAXS-restrained model is structurally compatible with the binding of the PERV intasome to BRD2. Altogether, these data confirm the important role of host BET proteins in the gamma-retroviruses' targeting site and efficiency of integration.

Porcine endogenous retrovirus-A/C: biochemical properties of its integrase and susceptibility to raltegravir.

Porcine endogenous retroviruses (PERVs) are present in the genomes of pig cells. The PERV-A/C recombinant virus can infect human cells and is a major risk of zoonotic disease in the case of xenotransplantation of pig organs to humans. Raltegravir (RAL) is a viral integrase (IN) inhibitor used in highly active antiretroviral treatment. In the present study, we explored the potential use of RAL against PERV-A/C. We report (i) a three-dimensional model of the PERV-A/C intasome complexed with RAL, (ii) the sensitivity of PERV-A/C IN to RAL in vitro and (iii) the sensitivity of a PERV-A/C-IRES-GFP recombinant virus to RAL in cellulo. We demonstrated that RAL is a potent inhibitor against PERV-A/C IN and PERV-A/C replication with IC50s in the nanomolar range. To date, the use of retroviral inhibitors remains the only way to control the risk of zoonotic PERV infection during pig-to-human xenotransplantation.

Dietary proteins contribute little to glucose production, even under optimal gluconeogenic conditions in healthy humans.

Dietary proteins are believed to participate significantly in maintaining blood glucose levels, but their contribution to endogenous glucose production (EGP) remains unclear. We investigated this question using multiple stable isotopes. After overnight fasting, eight healthy volunteers received an intravenous infusion of [6,6-²H2]-glucose. Two hours later, they ingested four eggs containing 23 g of intrinsically, uniformly, and doubly [¹5N]-[¹³C]-labeled proteins. Gas exchanges, expired CO2, blood, and urine were collected over the 8 h following egg ingestion. The cumulative amount of dietary amino acids (AAs) deaminated over this 8-h period was 18.1 ± 3.5%, 17.5% of them being oxidized. The EGP remained stable for 6 h but fell thereafter, concomitantly with blood glucose levels. During the 8 h after egg ingestion, 50.4 ± 7.7 g of glucose was produced, but only 3.9 ± 0.7 g originated from dietary AA. Our results show that the total postprandial contribution of dietary AA to EGP was small in humans habituated to a diet medium-rich in proteins, even after an overnight fast and in the absence of carbohydrates from the meal. These findings question the respective roles of dietary proteins and endogenous sources in generating significant amounts of glucose in order to maintain blood glucose levels in healthy subjects.

Postprandial protein metabolism but not a fecal test reveals protein malabsorption in patients with pancreatic exocrine insufficiency.

BACKGROUND & AIMS: Pancreatic exocrine insufficiency (PEI) impairs fat absorption, but few data are available on protein absorption. We investigated this question in patients with chronic pancreatitis, both in the absence and presence of enzyme therapy, using a stable isotope sensitive method. METHODS: Eleven patients with sustained PEI and regular enzyme substitution were investigated at hospital, after a washout period without enzyme substitution, and later after reintroduction of substitution. The digestibility and postprandial metabolism of dietary protein were characterized after the ingestion of a semi-synthetic single meal containing 20 g (15)N-labeled casein. RESULTS: At baseline, 20 ± 8% of dietary nitrogen was transferred to the metabolic pools vs. 24.5 ± 7% under enzyme treatment (P = 0.04). After treatment, the transfer of dietary nitrogen tended to increase in plasma amino acids, and increased significantly in plasma proteins and the deamination pool. In contrast, the fecal excretion of dietary nitrogen did not demonstrate any treatment effect. In patients not receiving insulin for diabetes, the treatment stimulated insulin secretion. CONCLUSIONS: Protein malabsorption was mostly undetectable using standard fecal tests. The study of the postprandial fate of dietary protein revealed a moderate increase of its transfer to metabolic pools after enzyme substitution.

The postprandial use of dietary amino acids as an energy substrate is delayed after the deamination process in rats adapted for 2 weeks to a high protein diet.

The aim of this study was to determine the contribution of dietary amino acids (AA) to energy metabolism under high protein (HP) diets, using a double tracer method to follow simultaneously the metabolic fate of α-amino groups and carbon skeletons. Sixty-seven male Wistar rats were fed a normal (NP) or HP diet for 14 days. Fifteen of them were equipped with a permanent catheter. On day 15, after fasting overnight, they received a 4-g meal extrinsically labeled with a mixture of 20 U-[(15)N]-[(13)C] AA. Energy metabolism, dietary AA deamination and oxidation and their transfer to plasma glucose were measured kinetically for 4 h in the catheterized rats. The transfer of dietary AA to liver glycogen was determined at 4 h. The digestive kinetics of dietary AA, their transfer into liver AA and proteins and the liver glycogen content were measured in the 52 other rats that were killed sequentially hourly over a 4-h period. [(15)N] and [(13)C] kinetics in the splanchnic protein pools were perfectly similar. Deamination increased fivefold in HP rats compared to NP rats. In the latter, all deaminated AA were oxidized. In HP rats, the oxidation rate was slower than deamination, so that half of the deaminated AA was non-oxidized within 4 h. Non-oxidized carbon skeletons were poorly sequestrated in glycogen, although there was a significant postprandial production of hepatic glycogen. Our results strongly suggest that excess dietary AA-derived carbon skeletons above the ATP production capacity, are temporarily retained in intermediate metabolic pools until the oxidative capacities of the liver are no longer overwhelmed by an excess of substrates.

[13C] GC-C-IRMS analysis of methylboronic acid derivatives of glucose from liver glycogen after the ingestion of [13C] labeled tracers in rats.

We developed a complete method to measure low [(13)C] enrichments in glycogen. Fourteen rats were fed a control diet. Six of them also ingested either [U-(13)C] glucose (n=2) or a mixture of 20 [U-(13)C] amino acids (n=4). Hepatic glycogen was extracted, digested to glucose and purified on anion-cation exchange resins. After the optimization of methylboronic acid derivatization using GC-MS, [(13)C] enrichment of extracted glucose was measured by GC-C-IRMS. The accuracy was addressed by measuring the enrichment excess of a calibration curve, which observed values were in good agreement with the expected values (R=0.9979). Corrected delta values were -15.6+/-1.6 delta(13)C (per thousand) for control rats (n=8) and increased to -5 to 8 delta(13)C (per thousand) per thousand and 12-14 delta(13)C (per thousand) per thousand after the ingestion of [U-(13)C] amino acids or [U-(13)C] glucose as oral tracers, respectively. The method enabled the determination of dietary substrate transfer into glycogen. The sequestration of dietary glucose in liver glycogen 4 h after the meal was 35% of the ingested dose whereas the transfer of carbon skeletons from amino acids was only 0.25 to 1%.

High-protein diets differentially modulate protein content and protein synthesis in visceral and peripheral tissues in rats.

OBJECTIVE: High-protein diets give rise to increased amplitude in the diurnal cycling of protein gains and losses at the whole-body level, but the tissue localization and mechanisms underlying these metabolic adaptations remain unclear. We investigated tissue-specific responses to increasing protein intakes in rats. METHODS: Protein synthesis rates (flooding dose with (13)C-valine) and accretion were assessed in individual tissues of fasted or fed rats (n = 32) after a 2-wk adaptation to a normal- or high-protein (HP) diet. RESULTS: In livers of HP rats, a strong inhibition of protein synthesis rates (-34%) occurred in the fasted and fed states, whereas a higher protein content (+10%) was observed. In the kidneys, a slight inhibition of synthesis rates after the HP diet was also observed but remained without effect on kidney protein pool size. Stomach and skin protein synthesis rates were significantly increased under HP conditions, whereas protein anabolism in skeletal muscle remained insensitive to the dietary protein level. This was also true for specific muscle protein fractions: myosin, mitochondrial, or sarcoplasmic protein synthesis rates were influenced by neither the dietary protein level nor the nutritional status. CONCLUSION: Modulation of protein kinetics and accretion by the HP diet is tissue-specific and the liver plays a critical role in such adaptations in a unique situation associating an inhibition of protein synthesis and protein pool expansion. The mechanisms underlying these changes and their physiologic incidence remain to be elucidated.

Ultra high temperature treatment, but not pasteurization, affects the postprandial kinetics of milk proteins in humans.

Although the chemical and physical modifications to milk proteins induced by technological treatments have been characterized extensively, their nutritional consequences have rarely been assessed in humans. We measured the effect of 2 technological treatments on the postprandial utilization of milk nitrogen (N), pasteurization (PAST) and ultra high temperature (UHT), compared with microfiltration (MF), using a sensitive method based on the use of milk proteins intrinsically labeled with (15)N. Twenty-five subjects were studied after a 1-wk standardization of their diet. On the day of the investigation, they ingested a single test meal corresponding to 500 mL of either MF, PAST, or UHT defatted milk. Serum amino acid (AA) levels as well as the transfer of (15)N into serum protein and AA, body urea, and urinary urea were determined throughout the 8-h postprandial period. The kinetics of dietary N transfer to serum AA, proteins, and urea did not differ between the MF and PAST groups. The transfer of dietary N to serum AA and protein and to body urea was significantly higher in UHT than in either the PAST or MF group. Postprandial deamination losses from dietary AA represented 25.9 +/- 3.3% of ingested N in the UHT group, 18.5 +/- 3.0% in the MF group, and 18.6 +/- 3.7% in the PAST group (P < 0.0001). The higher anabolic use of dietary N in plasma proteins after UHT ingestion strongly suggests that these differences are due to modifications to digestive kinetics and the further metabolism of dietary proteins subsequent to this particular treatment of milk.

Compared with casein or total milk protein, digestion of milk soluble proteins is too rapid to sustain the anabolic postprandial amino acid requirement.

BACKGROUND: The in vivo quality of milk protein fractions has seldom been studied in humans. OBJECTIVE: Our objective was to compare the postprandial utilization of dietary nitrogen from 3 [(15)N]-labeled milk products: micellar caseins (MC), milk soluble protein isolate (MSPI), and total milk protein (TMP). DESIGN: The macronutrient intakes of 23 healthy volunteers were standardized for 1 wk, after which time the subjects ingested a meal containing MC (n = 8), MSPI (n = 7), or TMP (n = 8). [(15)N] was measured for an 8-h period in plasma amino acids, proteins, and urea and in urinary urea. RESULTS: The transfer of dietary nitrogen to urea occurred earlier after MSPI ingestion than after MC and TMP ingestion, and concentrations remained high for 8 h, concomitantly with higher but transient hyperaminoacidemia and a higher incorporation of dietary nitrogen into plasma amino acids. In contrast, deamination, postprandial hyperaminoacidemia, and the incorporation of dietary nitrogen into plasma amino acids were lower in the MC and TMP groups. Finally, total postprandial deamination values were 18.5 +/- 2.9%, 21.1 +/- 2.8%, and 28.2 +/- 2.9% of ingested nitrogen in the TMP, MC, and MSPI groups, respectively. CONCLUSIONS: Our results confirm the major role of kinetics in dietary nitrogen postprandial utilization and highlight the paradox of MSPI, which, despite its high Protein Digestibility Corrected Amino Acid Score, ensures a rate of amino acid delivery that is too rapid to sustain the anabolic requirement during the postprandial period. Milk proteins had the best nutritional quality, which suggested a synergistic effect between soluble proteins and caseins.

Postprandial metabolic utilization of wheat protein in humans.

BACKGROUND: The quality of cereal protein has been little studied in humans despite its quantitative importance in the diet, particularly in developing countries. OBJECTIVE: The objective of this study was to determine the nutritional value of wheat protein in humans as assessed by the measurement of their real ileal digestibility and postprandial retention. DESIGN: Healthy young adults (n = 14) were fitted with an intestinal tube to allow the collection of intestinal fluid in the duodenum or terminal ileum. Subjects received a mixed meal of 136 g wheat toast that contained 24.6 g uniformly and intrinsically [(15)N]-labeled wheat protein. Intestinal fluid, blood, and urine were collected for 8 h postprandially. RESULTS: The real ileal digestibility of dietary wheat nitrogen amounted to 90.3 +/- 4.3%. The cumulative amount of dietary nitrogen transferred to the deamination pools reached a plateau at 8 h of 24.7 +/- 6.8% of the amount ingested. The urinary excretion of dietary nitrogen in ammonia was high (0.8 +/- 0.3% of ingested dose). The incorporation of dietary nitrogen into serum protein reached 7.0 +/- 1.9% of the meal. Postprandial wheat protein retention was 66.1 +/- 5.8%. CONCLUSIONS: Our results show that wheat proteins had the same true ileal digestibility as did most of the plant proteins already studied in humans, but also that they had a lower postprandial nitrogen retention value. However, this low value was higher than that predicted from the calculation of indispensable amino acid scores, ie, 89% rather than 30-40% of the nutritional value of milk proteins.

Increasing habitual protein intake accentuates differences in postprandial dietary nitrogen utilization between protein sources in humans.

It is appropriate to characterize the nutritional value of dietary proteins in humans through the specific study of dietary nitrogen metabolism during the postprandial period. However, the influence of the habitual protein intake on this variable has not been studied. We aimed to describe the influence of prior protein intake on the specific metabolic utilization of dietary nitrogen in humans. Healthy men and women were adapted for 7 d to two diets with a normal [NP, 1 g/(kg x d)] and high protein content [HP, 2 g/(kg x d)]. After each period, they were studied for an 8-h postmeal period after ingesting a single (15)N-labeled mixed meal (0.41 g/kg protein) containing either milk (n = 12) or soy protein (n = 8). The HP diet reduced the peak of dietary N incorporation into free serum amino acids in the soy group but had no effect in the milk group. The incorporation of dietary N into plasma protein was higher after soy than after milk protein, but habitual protein level had no effect. The postprandial retention of milk protein was reduced by the HP diet compared with the NP diet by only 5% and that of soy protein was diminished by 13% (protein source: P < 0.0001, protein level: P < 0.0001, interaction: P < 0.001). In conclusion, the efficiency of the meal N postprandial retention was lower after HP adaptation, but this decrease was much more pronounced for soy than for milk protein, indicating that increasing the habitual protein intake accentuates differences in metabolic utilization among dietary proteins.