Genetics: Is LADA just late onset type 1 diabetes?

Background: There is a controversy regarding Latent Autoimmune Diabetes in Adults (LADA) classification and whether it should be considered a slowly progressing form of type 1 (T1) diabetes (DM) or a distinct type of DM altogether. Methods: This cross-sectional study assessed major genes associated with T1DM (class II HLA, PTPN22 [rs2476601] and INS [rs689]) in patients with LADA, as compared with participants with T1DM (stratified according to age of diagnosis before or after 30) and T2DM. HLA genotyping of the DRB1, DQA1 and DQB1 loci was performed by reverse PCR sequence-specific oligonucleotides. HLA haplotypes were assigned according to those most frequently described in the European population. INS and PTPN22 SNPs were genotyped by real-time PCR. Results: A total of 578 participants were included: 248 with T1DM (70 diagnosed after the age of 30), 256 with T2DM and 74 with LADA. High risk HLA alleles were significantly more frequent in LADA than in T2DM, whereas the opposite was true for protective alleles. We found a lower frequency of the high-risk DRB1*04-DQB1*03:02-DQA1*03:01 haplotype in LADA (21.1%) than in the overall T1DM (34.7%) (p<0.05), whereas no differences were found between these groups for DRB1*03-DQB1*02:01-DQA1*05:01 or for protective alleles. Only 12% the overall T1DM group had no risk alleles vs 30% of LADA (p<0.0005). However, HLA allele distribution was similar in LADA and T1DM diagnosed after the age of 30. A total of 506 individuals (195 with T1DM [21 diagnosed after age 30] 253 with T2DM and 58 with LADA) were genotyped for the PTPN22 and INS SNPs. The G/A genotype of the PTPN22 rs2476601 was more frequent and the T/T genotype of the INS SNP rs689 was less frequent in T1DM compared to LADA. We did not find any significant differences in the frequency of the mentioned SNPs between LADA and T2DM, or between LADA and T1DM diagnosed after the age of 30. Conclusion: In this relatively small cross-sectional study, the genetic profile of subjects with LADA showed a similar T1DM-related risk allele distribution as in participants with T1DM diagnosed after the age of 30, but fewer risk alleles than those diagnosed before 30. Differences were present for HLA, as well as PTPN22 and INS genes.

Influence of storage time on pleural fluid adenosine deaminase activity.

BACKGROUND: Storing pleural fluid samples for research purposes is a common practice, but whether adenosine deaminase (ADA), an enzyme used for the diagnosis of tuberculous pleuritis, is stable over long periods of time is unknown. METHODS: We evaluated the stability of pleural ADA concentrations in 223 samples frozen at -800C as compared to values obtained immediately following the initial thoracentesis. Sample storage time ranged from several months to slightly more than 10 years. RESULTS: ADA activity was stable for up to 2.6 years. Afterwards, it decreased 6 to 8 U/L, enough to drop 2 (3.3%) tuberculous patients below the diagnostic ADA cutoff. CONCLUSIONS: As far as ADA enzymatic activity is concerned, pleural fluid samples are viable for extended periods of time. However, some caution in interpreting results from specimens stored for > 2.6 years is prudent.

Xpert® MTB/RIF in pleural fluid for the diagnosis of tuberculosis.

An automated nucleic acid amplification assay that simultaneously identifies Mycobacterium tuberculosis and rifampicin resistance, the Xpert® MTB/RIF test, has undergone extensive evaluation in sputum samples. Our aim was to define its diagnostic accuracy when performed on pleural fluid specimens. In 67 patients with pleural effusions, of whom half had tuberculous pleuritis, Xpert yielded 15% sensitivity and 100% specificity in the detection of tuberculosis (TB). Positive Xpert results tended to be more common in patients with microbiologically confirmed TB. Due to its low sensitivity, Xpert testing of pleural fluids has a limited role in the work-up of pleural effusions.

Comparison of polymorphonuclear- and lymphocyte-rich tuberculous pleural effusions.

SETTING: Most patients with tuberculous pleural effusions (TPE) have more than 50% lymphocytes in the pleural fluid. Data on patients in whom polymorphonuclear leukocytes (PMNLs) are the predominant cell type are scarce. OBJECTIVE: To compare the clinical, biochemical, microbiological and radiological characteristics between patients with predominantly PMNL and those with lymphocytic TPE. DESIGN: Retrospective analysis of 214 consecutive patients with TPE. RESULTS: The pleural fluid was PMNL-rich in 24 (11%) cases at the time of first thoracocentesis. Compared with those whose pleural fluid was predominantly lymphocytic, these patients showed a higher yield of mycobacteria in culture of sputum (50% vs. 25%, P = 0.03) and pleural fluid (50% vs. 10%, P < 0.01) on solid media, as well as higher pleural adenosine deaminase (ADA) levels (80 vs. 62 U/l, P = 0.02) at the expense of both ADA1 and ADA2 isoenzymes. A shift towards pleural lymphocytic predominance was observed in more than half of the PMNL-predominant patients subjected to repeat thoracocentesis. CONCLUSIONS: The finding of a predominantly PMNL exudate should not rule out TPE, particularly when pleural ADA activity is elevated. The collection of sputum and pleural fluid samples for mycobacterial culture should be encouraged in the case of suspected PMNL-rich TPE, as they are frequently positive in this early stage.

Resultado de un proceso para la mejora de las altas hospitalarias precoces / Process for improving early hospital discharge: a study of outcomes

Objetivo: Describir las intervenciones realizadas para acelerar la hora de alta hospitalaria y reducir la demora de ingreso de pacientes adultos médico-quirúrgicos desde urgencias y analizar los resultados alcanzados con la implementación del proceso. Método: Se analizó la situación del proceso de alta de nuestra organización y se identificaron las principales disfunciones. La reingeniería del proceso culminó con la elaboración del plan de alta hospitalaria para cuya implantación fueron necesarias intervenciones diversas sobre circuitos y sistemas de información. Los datos extraídos sobre las altas de pacientes adultos médico-quirúrgicos producidas en las diferentes franjas horarias (9 a 13, 13 a 17 y 17 a 9 horas), antes (2006) y durante los 2 años posteriores a la implementación (2008-2009) se analizaron mediante la prueba de (..) (AU)

Objectives: To describe process changes implemented to encourage early hospital discharge and reduce emergency admission delays for adult medical and surgical patients and to analyze outcomes achieved. Methods: Our hospital’s discharge system was analyzed to identify the main short comings. The process was then reengineered to create a hospital discharge plan whose implementation required several changes in care pathways and information systems. We gathered data on discharges of adult medical and surgical patients during 3 shifts (9 a.m. to 1P.M.; 1 to 5 P.M.; 5 to 9 P.M) before (2006) and during the 2 years (2008-2009) following the plan’s introduction in 2007. Outcomes were compared using the (..) (AU)

Biomarkers of infection for the differential diagnosis of pleural effusions.

We aimed to investigate whether pleural fluid concentrations of biomarkers for bacterial infection, namely triggering receptor expressed on myeloid cells (sTREM-1), procalcitonin (PCT), lipopolysaccharide-binding protein (LBP) and C-reactive protein (CRP), might identify infectious effusions and discriminate between complicated (CPPEs) and uncomplicated parapneumonic effusions (UPPEs). Stored pleural fluid samples from 308 patients with different causes of pleural effusion were used to measure the four biomarkers. Receiver-operating characteristic analysis determined the accuracy of the new tests. Median pleural fluid levels of CRP, sTREM-1 and LBP were significantly higher in CPPE compared with those in other aetiologies. The area under the curve for distinguishing infectious (parapneumonics and tuberculosis) from noninfectious effusions was 0.87 for CRP, 0.86 for sTREM-1, 0.57 for PCT and 0.87 for LBP. Regarding the discrimination of nonpurulent CPPE versus UPPE, a multivariate analysis found that pleural fluid glucose < or =60 mg x dL(-1), LBP > or =17 microg x mL(-1) and CRP > or =80 mg x L(-1) were the best parameters. Individually, none of the new biomarkers achieved better performance characteristics than pH, glucose or lactate dehydrogenase in labelling CPPE. In conclusion, elevated pleural fluid levels of CRP, sTREM and LBP identify patients with infectious effusions, particularly those with CPPE. PCT has no value for the differential diagnosis of pleural effusions.

Antinuclear antibody testing in pleural fluid for the diagnosis of lupus pleuritis.

We sought to determine whether measuring antinuclear antibodies (ANA) and their specificities [dsDNA, extractable nuclear antigens (ENA)] on pleural fluid may contribute to the differential diagnosis of pleural effusions. ANA were tested by indirect immunofluorescence on Hep-2 cells in the pleural fluid of 266 patients with effusions of different etiologies, including 15 lupus pleuritis. The cutoff value for diagnostic use was set at 1:160. Pleural fluid analysis of specific autoantibodies, such as anti-dsDNA and anti-ENA, was also performed if a positive ANA test was obtained. All patients with lupus pleurisy and 16 of 251 (6.4%) patients with pleural effusions secondary to other causes were ANA positive. Fifty-six percent of the positive ANAs in non-lupus pleural fluids were due to neoplasms. The pleural fluid ANA titers were low (< or = 1:80) or absent in two patients with systemic lupus erythematosus (SLE) and effusions due to other factors. Whereas ANA staining patterns in pleural fluid did not help to discriminate lupus pleuritis from non-lupus etiologies, the absence of pleural fluid anti-dsDNA or anti-ENA favored the latter. ANAs in pleural fluid provided no additional diagnostic information beyond that obtained by the measurement in serum and, therefore, these tests need not be routinely performed on pleural fluid samples. However, in patients with SLE and a pleural effusion of uncertain etiology, lack of ANAs or specific autoantibodies in pleural fluid argues against the diagnosis of lupus pleuritis.

Bayesian analysis using continuous likelihood ratios for identifying pleural exudates.

STUDY OBJECTIVES: To ascertain if equations that calculate continuous likelihood ratios (CLRs) for pleural exudates improve pleural fluid categorization, especially when false positive or false negative test results are obtained by using Light's criteria. DESIGN AND SETTING: Retrospective review of the clinical and pleural fluid data from a consecutive series of patients with pleural effusion who underwent thoracentesis at the University Hospital Arnau de Vilanova (Lleida, Spain) over an 11-year period. PATIENTS AND METHODS: A total of 1490 patients with pleural effusion (298 transudates and 1192 exudates) were recruited into the study. The presence of a transudate or exudate was established by clinical judgment. We examined the comparative diagnostic accuracy of 4 tests (i.e. pleural fluid protein and lactate dehydrogenase (LDH), and pleural fluid to serum protein and LDH ratios) for discriminating between transudates and exudates. Decision thresholds were determined by receiver operating characteristics (ROC) analysis. Equations for calculating CLRs derived from a logistic regression analysis based on a previously described method. RESULTS: Individual pleural fluid tests did not differ in their diagnostic accuracies according to ROC analysis. We calculated CLRs for the elements of Light's criteria and pleural fluid protein, and also illustrated the sequential use of CLRs for determining posttest probabilities. Overall, CLR formulas had marginal performance for the correct categorization of pleural fluid. CONCLUSIONS: CLRs provide a probabilistic statement as to the likelihood an effusion is a transudate or exudate. However, clinical judgment is little changed by the application of CLRs, and in doubtful cases a great amount of uncertainty remains. This Bayesian approach is likely to have no major impact on the clinical practice.

Differential effects of 2'-deoxyguanosine on peripheral blood mononuclear cell proliferation in healthy donors and Hashimoto's thyroiditis patients.

The aim of this study was to determine possible differences in peripheral blood mononuclear cells (PBMC) proliferation of healthy donors and Hashimoto's thyroiditis patients and whether a statistical approach to cell proliferation analysis might be used to discern the differences. The effect of a wide range of 2'-deoxyguanosine (dGuo) concentrations (0-1250 microM) on the mitogen-induced proliferation of PBMC was studied in healthy donors and Hashimoto's thyroiditis patients. Activity levels of purine nucleoside phosphorylase (PNP) and adenosine deaminase (ADA) in PBMC were also measured. For the first time in a study of these models of dGuo toxicity in vitro, the analysis of polynomial trends of orders from 1 to 7 was applied to evaluate cell proliferation. A dose-dependent inhibition of mitogen-induced PBMC proliferation was observed in both groups. Data for linear trend established that PBMC from Hashimoto's thyroiditis patients were more sensitive to dGuo toxicity than PBMC from healthy donors. A positive quadratic trend at low dGuo doses was found in the cell proliferation of Hashimoto's thyroiditis patients. A decrease in PNP activity (P < 0.025) and an increase in ADA activity (P < 0.005) was observed in PBMC of Hashimoto's thyroiditis group. The differences in PBMC proliferation subjected to dGuo toxicity between the two groups could be related with the distinct pattern of purine salvage enzymes observed.

Value of the determination of TNF-alpha in the plasma of patients with non-Hodgkins lymphoma.

The present report describes the value of the plasma determination of TNF-alpha, at diagnosis, in 43 patients with non-Hodgkin's lymphoma (NHL), related to their clinical presentation and the new International Prognosis Index (IPI). We also compare the levels of TNF-alpha with those of LDH, beta-2-microglobulin (beta-2-m), and ferritin. At diagnosis, the mean values of the quotients between the marker values and the maximum value of normal (ratio:r-) are placed 7 times higher than normal for r-TNF-alpha, whereas those of r-beta-2-m and r-LDH are 2,4 and 1,4 times more respectively. We found a relationship between the value of r-TNF-alpha and the ECOG, Ann Arbor stage, the number of affected extranodal sites, and between the values of r-beta-2-m with r-LDH. The best correlation was obtained between the values of r-TNF-alpha and r-beta-2-m and IPI, however r-TNF-alpha best stratify the four risk groups in this prognosis index.

Peripheral blood T-lymphocyte subsets in autoimmune thyroid disease.

Interest in T-lymphocyte subsets has arisen because of their involvement in the autoimmune process. Contradictory results have been published in the literature about the number of peripheral blood lymphocyte subsets in autoimmune diseases. In order to investigate the number and distribution of peripheral blood lymphocyte subsets in autoimmune thyroid disease, the levels of total T-lymphocytes (CD3), T-helper (CD4) and T-suppressor/cytotoxic (CD8) lymphocytes were determined in 44 patients with Graves' disease (1), multinodular goiter (2) and Hashimoto's thyroiditis (3). All patients had high levels of antithyroglobulin and thyroid antiperoxidase (antimicrosomal) antibodies. The T subset levels were related to the functional thyroid status, measured as serum free thyroxine (FT4) and thyrotropin (TSH). Our data show the existence of a strong influence of functional status on CD3, CD4 and CD8 levels, as reflected in the significant correlations obtained with FT4 (negative) and TSH (positive). A significant decrease in all populations was observed in Graves' disease hyperthyroid patients. A decrease in the CD4/CD8 ratio in Hashimoto's thyroiditis hypothyroid patients was observed, in contrast to an increase in the ratio in autoimmune hyperthyroid patients. This points to the CD4/CD8 ratio as a differential characteristic between the two autoimmune (hypothyroid and hyperthyroid) entities, independent of free thyroxine levels. No significant correlation was found between antithyroid antibody levels and peripheral blood T-lymphocyte subsets or serum levels of FT4 and TSH.

[Etomidate perfusion in neurosurgery]. / Perfusión de etomidato en neurocirugía.

In this prospective study, 20 patients undergoing mean duration (2-3 h) neurosurgical operations on fossa cranii posterior, and cervical and dorsolumbar rachis, were induced with 0.3 mg/kg etomidate bolus dose. To maintain anesthesia, etomidate perfusions at 10 micrograms/kg/min (group I) and 20 microg/kg/min (group II) were administered. Fentanyl at fractionated doses was used as analgesic without association to nitrous oxide and relaxation was achieved with pancuronium bromide. Quality of hypnosis, changes in serum concentration of cortisol as well as hemodynamic and electrolyte changes were evaluated. Serum concentrations of cortisol, glucose, sodium and potassium were measured in basal state, and during perioperative and postoperative period. Hemodynamic status was monitored and side effects were recorded. Patients of group I presented signs of deficient hypnosis consisting of marked sympathetic response. When etomidate perfusion ceased, both groups presented serum concentrations of cortisol under the lower normal limit. Six hours after operation, mean serum concentration of cortisol in group 2 patients was significantly lower than in group I patients; in their samples, a dose-dependent recovery was detected. Serum glucose concentration increased during anesthesia and operation and serum electrolyte concentration remained within normal range during perioperative and postoperative periods. Our results demonstrate that a 20 micrograms/kg/min etomidate infusion is adequate to achieve neuroanesthesia when nitrous oxide administration is contraindicated.