An IDH1-vitamin C crosstalk drives human erythroid development by inhibiting pro-oxidant mitochondrial metabolism.

The metabolic changes controlling the stepwise differentiation of hematopoietic stem and progenitor cells (HSPCs) to mature erythrocytes are poorly understood. Here, we show that HSPC development to an erythroid-committed proerythroblast results in augmented glutaminolysis, generating alpha-ketoglutarate (αKG) and driving mitochondrial oxidative phosphorylation (OXPHOS). However, sequential late-stage erythropoiesis is dependent on decreasing αKG-driven OXPHOS, and we find that isocitrate dehydrogenase 1 (IDH1) plays a central role in this process. IDH1 downregulation augments mitochondrial oxidation of αKG and inhibits reticulocyte generation. Furthermore, IDH1 knockdown results in the generation of multinucleated erythroblasts, a morphological abnormality characteristic of myelodysplastic syndrome and congenital dyserythropoietic anemia. We identify vitamin C homeostasis as a critical regulator of ineffective erythropoiesis; oxidized ascorbate increases mitochondrial superoxide and significantly exacerbates the abnormal erythroblast phenotype of IDH1-downregulated progenitors, whereas vitamin C, scavenging reactive oxygen species (ROS) and reprogramming mitochondrial metabolism, rescues erythropoiesis. Thus, an IDH1-vitamin C crosstalk controls terminal steps of human erythroid differentiation.

Tumor markers in pancreatic cystic fluids for diagnosis of malignant cysts.

BACKGROUND AND AIM: Early diagnosis of premalignant or malignant pancreatic cysts is essential to improve prognosis. Sampling of pancreatic cyst fluid by fine-needle aspiration during endoscopic ultrasonography (EUS) enables cytopathological examination combined with biochemical analysis. This study aimed to provide an aid based on biological markers for the preoperative management of patients with pancreatic cysts. METHODS: Pancreatic fluids obtained by EUS-guided fine-needle aspiration from 115 patients with cystic lesions were assayed for amylase, lipase, carcinoembryonic antigen (CEA), CA 19-9 and CA 72-4. In addition, chromogranin A (CgA) and neuron-specific enolase (NSE) were measured in 28 fluid samples. RESULTS: ROC curve analysis of the different markers for diagnosis of mucinous cysts showed that CEA had the highest area under the curve (0.93, 95% CI 0.87-0.97), with a sensitivity and specificity of 89% and 93%, respectively, at the cutoff value of 317 µg/L. The CgA and NSE concentrations in 5 NET cysts (median values of 210 [63-492] and 68.5 [9-496] µg/L, respectively) were higher than in 23 other cysts (median values of 8 [7-828] and 2.7 [0.5-35.8] µg/L, respectively) (p = 0.0015 and p = 0.0045, respectively). CONCLUSIONS: CEA is the best marker for identifying a cyst as mucinous. In case of low levels of CEA, our results suggest that CgA and NSE measurements may be helpful in the diagnosis of a neuroendocrine tumor and therefore deserve further investigation.

TMPRSS2-ERG fusion transcripts expression in patients referred for prostate biopsy: combining detection in urine and needle rinse material.

OBJECTIVES: The objective of this study was to combine urine and prostate biopsy rinse material (BRM) assays to increase sensitivity for fusion gene detection. PATIENTS AND METHODS: A total of 194 patients with suspicion of prostate cancer were prospectively included. Urine samples were collected before or after prostate biopsy, as well as BRM. RT-qPCR was used for the detection of fusion transcripts. A microfocal cancer on biopsy was defined by a single core involved with less than 3 mm of Gleason score 3 + 3 cancer. The association between RT-qPCR and biopsy results was statistically assessed. RESULTS: Seven patients were excluded because of insufficient material. Cancer was detected on biopsy in 100 (53%) patients. Urine alone, BRM alone and both samples were obtained in 155, 164 and 132 patients, respectively. In patients with evidence of cancer on biopsy, a fusion transcript was detected in 63, 55 and 73% of the cases on urine alone, BRM alone and paired samples, respectively. Fusion gene detection on BRM was only associated with the amount of cancer on biopsy. Urine fusion score had a larger area under the curve than serum PSA (p = 0.002) and was significantly higher in patients with high Gleason score and significant cancer on biopsy. Assays of paired samples allowed increasing sensitivity in all subgroups of patients. CONCLUSIONS: TMPRSS2-ERG fusion gene detection may be performed both in the urine and BRM to increase sensitivity. However, only T-E urine score was associated with adverse pathological features.

Automated two-site immunofluorescent assay for the measurement of serum chromogranin A.

OBJECTIVES: Chromogranin A (CgA) is the best-characterized biological marker common to neuroendocrine tumours and is therefore recommended for their diagnosis. The measurement of serum CgA is of great importance for reaching an early diagnosis and thus reducing the delay before treatment is instigated. The Kryptor CgA assay is the first fully automated assay available. The aim of this study was to evaluate its analytical performance. DESIGN AND METHODS: The imprecision and linearity of the Kryptor CgA assay were evaluated. This assay was compared with the Cis Bio CgA RIA assay in 78 serum samples. Its clinical utility was assessed in serum from 229 patients. RESULTS: The study performed on imprecision of Kryptor measurements showed intra- and inter-run CVs ≤ 5%. The study of linearity showed a satisfactory recovery rate for CgA concentrations up to 1200 µg/L. The Kryptor and RIA assays agreed well on the basis of the cut-off values provided by the two manufacturers. The Bland and Altman plot of the values obtained (range: 20-5560 µg/L) provided a mean difference of -10.1 µg/L (SD: 116). The clinical sensitivities of Kryptor CgA for diagnosis of pheochromocytoma and paraganglioma (n 20) and gastroenteropancreatic NETs (n 17) were respectively 100 and 94%. CONCLUSIONS: The Kryptor assay for CgA shows reliable analytical and clinical characteristics and allows a fast delivery of results.

Interleukin-10 promoter (-1082) polymorphism in association with repeated hospital-acquired infections in elderly patients.

BACKGROUND: Infections are frequent complications of hospitalization, particularly in the elderly. Pro- and anti-inflammatory cytokines are essential components of the host response to pathogens and polymorphisms in their genes may contribute to inter-individual variations of the inflammatory response. The aim of this study was to investigate whether cytokine polymorphisms, separately or in combination, could be determining factors in the development of repeated nosocomial infections in elderly hospitalized patients. METHODS: Tumor necrosis factor-α (-308) and (-238), interleukin-6 (-174) and (-6331), interleukin-10 (-1082) and (-592) polymorphisms were genotyped by PCR and hybridization with fluorescent-labeled probes in 245 hospitalized elderly patients (mean age 85.2 years; SD 6) and compared with those in 145 healthy adults. RESULTS: The distribution of genotypes did not differ between elderly patients and control subjects. The presence of the interleukin-10 A(592) or A(1082) allele was more frequent individually and after adjustment for multiple comparisons in patients who suffered from several infections (p = 0.012, odds ratio = 5.3; 95 % confidence interval = 1.2-23.1). CONCLUSION: Our data support a determinant role for interleukin-10 (-1082) polymorphism in the development of nosocomial infections.

TMPRSS2-ERG fusion transcripts in matched urine and needle rinse material after biopsy for the detection of prostate cancer.

BACKGROUND: Current methods for detecting TMPRSS2-ERG fusion transcript in the urine of patients with suspected prostate cancer lack diagnostic sensitivity. We combined urine and prostate biopsy rinse material (BRM) assays to improve the fusion gene detection rate. METHODS: Eighty patients with clinical and/or prostate-specific antigen suspicion of prostate cancer were prospectively included in the study. Urine samples were collected before and after prostate biopsy, and BRM was collected from the biopsy needle. We used reverse-transcription PCR (RT-PCR) for the detection of fusion transcripts. Microfocal cancer (MFC) on biopsy was defined by a single core involved with ≤3 mm of cancer with Gleason score 3 + 3. We statistically assessed the association between RT-PCR and biopsy results. RESULTS: Urine alone, BRM alone, and both samples were obtained in 4, 19, and 57 patients, respectively. Three patients were excluded because of insufficient material. In the remaining 77 patients, cancer was detected on biopsy in 42 (55%). The diagnostic sensitivity of the assay for cancer detection was 62% (95% CI 47%-78%), 69% (53%-85%), and 89% (73%-99%) with BRM alone, urine alone, and paired samples, respectively. The lowest values were obtained with the urine assay in patients with MFC or Gleason score >3 + 3 cancer. Assays of paired samples provided increased diagnostic sensitivity in all subgroups of patients. CONCLUSIONS: TMPRSS2-ERG fusion gene detection may be improved by performing assays in both urine and BRM. Insufficient cell numbers in urine samples and cell lysis during centrifugation may explain the low diagnostic sensitivity of the urine assay.

Impact of comorbidities on hospital-acquired infections in a geriatric rehabilitation unit: prospective study of 252 patients.

OBJECTIVES: Hospital-acquired infections (HAIs) remain a major source of morbidity and mortality in long-term care units, despite advances in antimicrobial therapy and preventive measures. Our aim was to investigate risk factors for HAIs, especially in the elderly, and to describe the relationship between comorbidities (number, severity, and specific diseases) and HAIs using a comprehensive inventory of comorbidities. DESIGN: Prospective cohort study SETTING: Geriatric rehabilitation unit in a university hospital in the Paris metropolitan area. PARTICIPANTS: Participants were 252 consecutive patients aged 75 years or older (mean age, 85 ± 6.2 years) and admitted between 2006 and 2008. MEASUREMENTS: Surveillance of HAI was conducted. A complete inventory of comorbidities was done using the Cumulative Illness Rating Scale for Geriatrics (CIRS-G). Potential risk factors were evaluated in 2 risk models, one with HAI acquisition, CIRS-G, activities of daily living score less than 10, and at least 1 invasive procedure (yes/no) and the other with HAI acquisition and specific invasive procedures and diseases. RESULTS: Of the 252 patients, 97 experienced HAIs, for an incidence of 5.6 infections per 1000 bed-days. The most common HAI sites were the respiratory tract (48%; 65/136) and urinary tract (37%; 51/136). The CIRS-G global score and comorbidity index were higher in patients with than without HAIs. Among HAI categories, respiratory and urogenital diseases were more prevalent in the group with HAIs. In the model combining CIRS-G, activities of daily living score less than 10, and at least 1 invasive procedure, independent risk factors for HAI were CIRS-G index (odds ratio [OR], 1.55; 95% confidence interval [95% CI], 1.13-2.11; P = .005) and invasive procedures (OR, 5.18; 95% CI, 2.77-9.71; P < .001). In the model including specific procedures and diseases, independent risk factors for HAI were intravenous catheter (OR, 7.39; 95% CI, 2.94-18.56; P < .001), urinary catheter (OR, 3.33; 95% CI, 1.40-7.88; P = .006), gastrointestinal endoscopy (OR, 3.69; 95% CI, 1.12-12.16; P = .03), pressure sores (OR, 2.52; 95% CI, 1.04-6.10; P = .03), and swallowing impairment (OR, 3.37; 95% CI, 1.16-9.74; P = .02). CONCLUSIONS: This study identified several important risk factors for HAIs. There is a need for HAI prevention via the implementation of infection-control programs, including surveillance, in rehabilitation units.

Porous metal-organic-framework nanoscale carriers as a potential platform for drug delivery and imaging.

In the domain of health, one important challenge is the efficient delivery of drugs in the body using non-toxic nanocarriers. Most of the existing carrier materials show poor drug loading (usually less than 5 wt% of the transported drug versus the carrier material) and/or rapid release of the proportion of the drug that is simply adsorbed (or anchored) at the external surface of the nanocarrier. In this context, porous hybrid solids, with the ability to tune their structures and porosities for better drug interactions and high loadings, are well suited to serve as nanocarriers for delivery and imaging applications. Here we show that specific non-toxic porous iron(III)-based metal-organic frameworks with engineered cores and surfaces, as well as imaging properties, function as superior nanocarriers for efficient controlled delivery of challenging antitumoural and retroviral drugs (that is, busulfan, azidothymidine triphosphate, doxorubicin or cidofovir) against cancer and AIDS. In addition to their high loadings, they also potentially associate therapeutics and diagnostics, thus opening the way for theranostics, or personalized patient treatments.

Validation of body mass index for the diagnosis of malnutrition in patients with liver cirrhosis.

OBJECTIVES: Simple clinical tools are needed to detect malnutrition in cirrhotic patients. We have validated optimal body mass index (BMI) cut-offs for the diagnosis of malnutrition in accordance with the importance of ascites. METHODS: BMI, mid-arm muscle circumference (MAMC) and triceps skinfold thickness (TST) were measured before paracentesis in study (SP) and validation (VP) populations of 875 and 294 cirrhotic patients, respectively with no ascite (NA), mild (MA), tense ascites (TA) (NA/MA/TA: SP: 327, 270, 278; VP: 111, 69, 114). Preserved nutritional status (SP: 259; VP: 93), malnutrition including severe and moderate malnutrition (SP: 251 and 365; VP: 92 and 109) were defined from MAMC and TST measurements. RESULTS: Optimal BMI cut-off values were 22, 23 and 25 kg/m2 in NA, MA and TA patients, respectively. In the whole SP and VP, sensitivities of these cut-offs were 86.2% and 89.1%, respectively; the corresponding negative predictive values (NPV) for the diagnosis of severe malnutrition were 92.3% and 93.2%; specificities and positive predictive values (PPV) were 87.7% and 89.9%, 92.7% and 93.6%, respectively for the diagnosis of malnutrition. In the entire VP, peripheral oedema did not change sensitivity and NPV of BMI cut-offs for the diagnosis of severe malnutrition and tended to increase specificity and PPV for the diagnosis of malnutrition. 96.7% of the malnourished TA patients in the VP before paracentesis were correctly identified after removal of ascites. CONCLUSION: BMI is a reliable parameter to detect malnutrition in cirrhotic patients with the above mentioned cut-offs. Peripheral oedema and removal of ascites do not affect its diagnostic performance.

Combined polymorphisms of tumour necrosis factor alpha and interleukin-10 genes in patients with alcoholic hepatitis.

OBJECTIVES: The development and progression of alcoholic hepatitis are controlled by an extensive cytokine network which involves pro-inflammatory and anti-inflammatory cytokines. Genetic variations determining production of these cytokines have been described and the susceptibility to the disease may be determined by an imbalance in the expression of several candidate genes. METHODS: We have studied biallelic single nucleotide polymorphisms at positions (-308) and (-238) in the promoter region of the pro-inflammatory tumour necrosis factor alpha (TNF-alpha) and at positions (-1,082) and (-592) in the promoter of anti-inflammatory interleukin-10 (IL-10) in 134 patients with severe biopsy-proven alcoholic hepatitis and 145 healthy subjects. RESULTS: The frequency distribution of isolated cytokine genotypes did not differ between the two groups. The combination of at least one A or A allele for TNF-alpha, associated with a TNF-alpha high-producer phenotype, and one A or A allele for IL-10, associated with an IL-10 low-producer phenotype, was less frequent in patients (20.9 vs 33.8%, P=0.016, OR (95% CI)=0.52 (0.30-0.89)). The same combination in patients was associated with a higher risk of septic complications (32.5 vs 16.0%, P=0.031, OR (95% CI)=1.79 (1.07-6.00)) but not with in-hospital mortality. CONCLUSIONS: We have not found any relationship between the isolated polymorphisms and the risk of alcoholic hepatitis. Moreover, the imbalance between the pro-inflammatory and anti-inflammatory responses leading to high TNF-alpha production and low IL-10 was uncommon in alcoholic hepatitis. However, patients with this particular genotype appeared more susceptible to severe septic complications.

Relations between undernutrition and nosocomial infections in elderly patients.

BACKGROUND: Hospital-acquired infections and malnutrition are of major concern in public health in elderly patients. However, the interactions between these two entities are not well established. OBJECTIVES: To determine the incidence of nosocomial infections (NI) and its association with malnutrition. SUBJECTS: 185 hospitalised older adults aged 81.6+/-0.6 years old were nutritionally assessed on admission by measurement of anthropometric variables, serum nutritional proteins and evaluation of dietary intake. During hospitalisation, patients' progress was closely monitored, particularly for the detection of nosocomial infections. RESULTS: The incidence rate of NI was 59% and the global infection rate was 7.6/1,000 bed days. The most common infection site was the urinary tract (n=63). The nutritional status of the population was studied by comparing three groups defined according to the absence (group I, n=116), presence of one infection (group II, n=38) or presence of more than one infection (group III, n=31). All but one anthropometric parameters varied among the three groups. Total energy intake also varied among the three groups. The group I had higher daily nutrient intake than the other two groups (respectively P=0.004 and P<0.0001). Albumin, transthyretin, and C-reactive protein levels differed significantly among the three groups (respectively P<0.0001, P<0.0001 and P=0.0003). Age, energy intake, length of hospital stay and the presence of a urinary catheter were independent risk factors of nosocomial infection. CONCLUSION: Our findings show that patients with multiple NI were older, showed an altered nutritional status, a prolonged recovery, more frequently had urinary catheters and more discharge placement.

Enteral nutrition in severely malnourished and anorectic cirrhotic patients in clinical practice.

OBJECTIVES: To determine among severely malnourished cirrhotic patients remaining anorectic during hospital stay which patients may benefit from enteral nutrition in clinical practice. METHODS: A prospective study including malnourished cirrhotic patients fed by enteral nutrition because of inadequate dietary intake after one-month hospitalization was carried out in a department receiving patients from other hospitals. Patients who died during hospital stay (N=35, group I) were compared to surviving patients (N=28, group II). RESULTS: Nutritional status and spontaneous dietary intake on admission to our department were in the same range in the two groups, Pugh score was higher in group I (11.1 +/- 1.9 vs 9.1 +/- 2.0, P=0.0001). The delay between previous hospital admission and the outset of enteral nutrition was comparable in the two groups. Its duration and total dietary intake during enteral nutrition were higher in group II (respectively 42.2 +/- 30.9 vs 15.2 +/- 33.1 days, P=0.0016 and 41.1 +/- 13.0 vs 29.9 +/- 10.0 kcal/kg/d, P=0.0004). Prevalence of side effects was higher in group I (54.3 vs 17.9%, P=0.0031). Multivariate analysis showed that Pugh score and septic complications were negatively associated with survival (respectively P=0.0196 and P=0.0078) while duration of enteral nutrition was positively associated (P=0.0435). Eighty six per cent of patients receiving enteral nutrition with bilirubin levels above 74 micromol/L on admission to our department died during hospital stay. Mid-term effects of enteral nutrition in surviving patients were improvement in Pugh score (7.5 +/- 2.0 vs 9.1 +/- 2.0, P<0.0001) and increase in spontaneous caloric and protein intake (29.7 +/- 15.3 vs 18.1 +/- 10.1 kcal/kg/d, P=0.0150 and 1.0 +/- 0.5 vs 0.6 +/- 0.3 g/kg/d, P=0.0049). CONCLUSIONS: In severely malnourished cirrhotic patients remaining anorectic after one-month hospitalization, patients with bilirubin level below 74 micromol/L may benefit from six-week enteral nutrition with mid-term improvement in liver function and increase in spontaneous dietary intake.

Oral candidiasis and nutritional deficiencies in elderly hospitalised patients.

The prevalence of oral candidiasis and its association with malnutrition in terms of protein-energy malnutrition and mineral and vitamin depletion were evaluated in ninety-seven hospitalised older adults aged 82.1 (SD 8.6) years. Patients underwent a complete oral examination with microbiological investigation on admission to our geriatric rehabilitation unit. Patients were assessed nutritionally by evaluation of dietary intake and measurement of anthropometric variables, serum nutritional proteins, ferritin, Zn, folate, vitamins B12 and C. The prevalence of oral candidiasis was 37% (n 36); the proportion of patients with BMI <20 kg/m(2) was 32% (n 31). The nutritional status of the population was studied by comparing two groups defined according to the absence (group I; n 61) or presence (group II; n 36) of oral candidiasis. The two groups did not differ on the basis of BMI and mid-arm circumference. However, group II had a smaller leg circumference, lower daily energy and protein intakes, lower albumin and transthyretin levels. Patients successfully treated with fluconazole increased their intake on day 30. The proportion of patients with hypozincaemia (<12.5 micromol/l) and vitamin C deficiency (<0.7 mg/l) was higher in group II. Treatment with antibiotics, poor oral hygiene, denture wearing, and vitamin C deficiency appeared as the most significant independent risk factors associated with oral candidiasis. The present findings show that oral candidiasis appears to be related to malnutrition and results in mucosal lesions that have a negative impact on energy intake, which may subsequently worsen nutritional status.

Evaluation of nutritional practice in hospitalized cirrhotic patients: results of a prospective study.

OBJECTIVE: We evaluated the impact of malnutrition and nutrition practice in 396 hospitalized cirrhotic patients (Child stages A, B, and C: 60, 169, and 167 patients) in a readaptative unit for liver diseases. METHODS: Triceps skinfold thickness and mid-arm muscle circumference were measured at admission, and spontaneous dietary intake was evaluated at admission and during hospital stay. RESULTS: Ascites was associated with impairment of nutrition status: 49% and 30.4% of non-ascitic patients, 49.1% and 40.5% of patients with mild ascites, and 65.5% and 48.3% of patients with tense ascites had mid-arm muscle circumferences and triceps skinfold thicknesses, respectively, below the fifth percentile of a reference population (P < 0.05 and P = 0.02, respectively). Decrease in dietary intake paralleled worsening of liver failure: 48% and 34% of Child A patients, 51.7% and 35.8% of Child B patients, and 80.3% and 62.9% of Child C patients at admission had caloric intakes below 30 kcal/kg of body weight and protein intakes below 1 g/kg of body weight, respectively (P < 0.001). Mortality rate was 15.4% during hospital stay; Child-Pugh score (P = 0.0014), age (P = 0.0029), severe septic complications (P = 0.0050), and decrease in caloric intake during hospital stay (P = 0.0072) were independently associated with mortality. Twenty-four patients received enteral feeding that was initiated before admission in four patients and after 12.4 +/- 8.3 d of hospitalization in 20 patients because of low caloric intake (<25 kcal/kg) despite oral supplements. Patients receiving enteral feeding were older (P < 0.01), had a higher Child-Pugh score (P < 0.01), and a higher mortality rate (P < 0.001) than other patients. CONCLUSIONS: Hospitalized cirrhotic patients have a high prevalence rate of malnutrition, and most do not satisfy their nutritional requirements. Decrease in caloric intake is an independent risk factor of short-term mortality. Enteral nutrition after failure of oral supplementation has no clinical benefit. Tube feeding may be indicated earlier in the course of the disease.

[Prognosis factors of short and long-term survival in elderly hospitalized patients after percutaneous endoscopic gastrostomy]. / Facteurs pronostiques de la survie à court et long terme après pose d'une gastrostomie percutanée endoscopique chez des malades âgés hospitalisés.

OBJECTIVE: Percutaneous endoscopic gastrostomy (PEG) is the procedure of choice to achieve long-term enteral nutrition. The risks and benefits of PEG in elderly hospitalized patients have been poorly documented. The objective of this study was to describe the outcome of elderly patients one-year after insertion of a PEG tube. PATIENTS AND METHODS: Hospital records of 73 patients who underwent PEG for enteral nutrition were reviewed retrospectively. Data on patient age and sex, preexisting medical conditions such as dementia or pressure sores, indication for PEG, concomitant infection, complications of PEG and death were obtained from the hospital charts. RESULTS: The main indication for PEG was anorexia (49%). Before insertion of the gastrostomy tube, 44% of the patients had pressure scores, 30% had concomitant infection, 45% had dementia. PEG complications were observed in 51 patients. The survival rate at 1, 6 and 12 months was 0.68 [95% confidence interval - CI 95%: 0.56-0.78], 0.48 [CI 95%: 0.36-0.59] and 0.37 [CI 95%: 0.26-0.48] respectively. The presence of an infectious disease or of pressure sores at the time of PEG tube insertion were independently associated with mortality. Median survival of patients with these two factors was 32 days [CI 95%: 11-98]. CONCLUSION: According to these results, the PEG tubes should be inserted with a delay from infectious diseases and before the occurrence of pressure sores.