Brucella spondylitis with paravertebral abscess due to Brucella melitensis infection: a case report

This report describes the case of a 45-year-old woman with a 5-month history of fever, generalized malaise, myalgia, lower back pain and difficulty in walking. Serodiagnosis for brucella, carried out at the onset of symptoms 5 months previously, was negative. When the patient was admitted to our hospital there was contracture of the paraspinal muscles but no peripheral nerve damage. Laboratory tests showed positive agglutination for Brucella and an increase in the rate of dilution from 1/160 to 1/640 over 2 weeks. Radiographs and a computed tomography scan of the spine revealed bone erosion in the posterior borders of the L4-L5 vertebral end plates and a soft tissue mass surrounding the interposed disc and protruding into the spinal canal. Magnetic resonance imaging confirmed the presence of a paraspinal abscess around the affected disc and tissue edema. Culture tests of the blood and abscess tissue, taken by biopsy, were negative. Rifampicin treatment (600 mg daily), combined with a bust cast to immobilize the spine, led to clinical healing without the need for surgery. Because onset symptoms are nonspecific and insidious, in nonrisk subjects a diagnosis of brucellosis may sometimes be suspected only if there are local symptoms. The phenomenon of the absence of positivity in patients with a high antibody titer should also be considered Cases such as that described herein demonstrate the need for culture tests and serodiagnosis, even in nonrisk patients with persistent fever and arthralgia, to prevent the later complications of brucellosis.     

A proposal of new ocular items in Sjögren's syndrome classification criteria

OBJECTIVE: To verify whether ocular surface tests other than those included in primary Sj?gren's syndrome (SS-I) classification criteria (Schirmer I, Break up Time, vital dye staining) may contribute to SS I diagnosis. METHODS: Two hundred and sixty-two patients (78 SS-1, 91 non-SS autoimmune diseases, 93 Sicca syndrome) filled a validated questionnaire on symptoms and were evaluated by Schirmer test without (Schirmer I) and with (Jones test) topical anaesthesia, Break Up Time (BUT), corneal aesthesiometry, tear clearance rate, vital dye (lissamine green) staining, impression conjunctival cytology, concentration of tear lysozyme and lactoferrin. Thresholds were selected from Receiver Operating Curves; sensitivity, specificity, likelihood ratio (LR+), predictive values were calculated for each test. A logistic regression model was constructed representing the best diagnostic index for SS. RESULTS: Data showed a poor diagnostic performance of Schirmer test I (LR+ 1.38) and BUT (LR+ 1.05); results from lissamine green staining may be unreliable due to incorporation bias. Tear lactoferrin (LR+ 4.52), Jones test (LR+ 6.24), tear lysozyme (LR+ 8.0), symptom questionnaire (LR+ 8.62), tear clearance rate (LR+ 18.73) and corneal aesthesiometry (LR+ 20.96) exhibited high diagnostic performance also taken together in the regression model. CONCLUSION: Because many of the tests we have screened in this study can be carried out by a trained ophthalmologist in any clinical setting, we recommend that ocular surface impairment is studied with the combination of tests proved to be helpful for the SS I diagnosis.     

Diagnostic performance of tear function tests in Sjogren's syndrome patients

OBJECTIVE: To evaluate the diagnostic performance of the tests included in primary Sjogren's syndrome (SS-I) diagnostic criteria (Schirmer I, break-up time, vital dye staining) and to compare them with other examinations related to the ocular surface status. METHODS: Clinical and cytological data were collected from 177 patients (62 SS-1, 56 non-SS autoimmune diseases, 59 Sicca syndrome). Tear tests included: a validated questionnaire on symptoms, Schirmer I, Jones test, Ferning test, BUT, corneal aesthesiometry, tear clearance test, lissamine green staining, impression conjunctival cytology. Data were statistically evaluated and sensitivity, specificity, likelihood ratio (LR+), receiver-operating characteristics (ROC) curves were calculated for each test. RESULTS: Data showed a poor diagnostic performance of Schirmer test I (sensitivity 0.42; specificity 0.76; LR+1.75) and BUT (sensitivity 0.92; specificity 0.17; LR+1.11) (area under the curve in ROC analysis <0.58). Validated subjective symptoms questionnaire (sensitivity 0.89; specificity 0.72; LR+3.18), Jones test (sensitivity 0.60; specificity 0.88; LR+5), corneal aesthesiometry (sensitivity 0.80; specificity 0.67; LR+2.42), and tear clearance test (sensitivity 0.63; specificity 0.84; LR+3.93), all exhibited a high diagnostic performance (area under the curve in the ROC analysis always >0.70). Lissamine green staining exhibited the best performance (sensitivity 0.63; specificity 0.89; LR+5.72) but the result could be distorted by an incorporation bias. CONCLUSIONS: Our data suggest to implement the items for ocular signs and symptoms contained in many SS-I diagnostic criteria with the use of a validated questionnaire, performance of Jones test, corneal aesthesiometry measurement, and tear clearance rate evaluation.     

Modulation of human longevity by SIRT3 single nucleotide polymorphisms in the prospective study “Treviso Longeva (TRELONG)”

Human sirtuins are seven proteins with deacetylase activity that are emerging as key modulators of basic physiological functions. Some evidence links SIRT3 to longevity in mammals. This study aimed to investigate whether variants within SIRT3 gene were associated to human longevity. We analyzed 549 genomic DNA collected during the prospective study “Treviso Longeva,” including elderly over 70 years of age from the municipality of Treviso, a small city in the northeast of Italy. We genotyped SIRT3 rs3825075, rs4980329, and rs11555236 single nucleotide polymorphisms (SNPs) by real-time polymerase chain reaction allelic discrimination assay. A cross-sectional analysis performed by comparing people over and under 85 years of age did not evidence association among the SIRT3 SNPs and longevity. However, when we performed a longitudinal analysis considering mortality as a dependent variable, we observed an association of SIRT3 rs11555236 and rs4980329 with longevity in the whole population (p values corrected for potential confounders = 0.04 and 0.03, respectively). After stratification according to gender, the same SNPs were associated to female longevity only (p values corrected for potential confounders = 0.03 and 0.02, respectively). Finally, as rs11555236 was reported to be in linkage disequilibrium with a putative functional enhancer within the SIRT3 gene, we assessed whether rs11555236 genotypes correlated with a different level of SIRT3 protein in peripheral blood mononuclear cells. We found an increased level of SIRT3 in subjects homozygous for the (T) allele. We suggest that SIRT3 genetic variability might be relevant for the modulation of human longevity in the Italian population.

Electronic supplementary material

The online version of this article (doi:10.1007/s11357-013-9559-2) contains supplementary material, which is available to authorized users.     

The relationship of vitamin D status to bone mineral density in an Italian population of postmenopausal women

Several authors have found a relationship between vitamin D status and bone mineral density (BMD). To our knowledge, no previous studies on this topic have been carried out on the Italian postmenopausal population. We studied this relationship retrospectively in 156 Italian postmenopausal women. We also investigated the relationship between parathyroid hormone (PTH) and BMD. Measurements of BMD were taken at the lumbar spine and upper femur by dual X-ray absorptiometry. Serum 25(OH)D (calcidiol), 1,25(OH)2D (calcitriol), PTH, calcium, phosphorus, creatinine, osteocalcin and urinary calcium and phosphorus were measured according to the current laboratory methods of analysis. We found a positive statistically significant correlation between BMD, both at the spine and hip, and 25(OH)D, and a negative statistically significant correlation between BMD and PTH. No statistically significant correlation was found between BMD and 1,25(OH)2D. Crude logistic regression showed age, 25(OH)D and PTH were significant predictors of low BMD, while 1,25(OH)2D was not. Backward logistic regression showed 25(OH)D was the best predictive model for spine osteoporosis together with age, and on its own it was the best predictive model for femoral neck osteoporosis.No funding sources supported this publication.     

Substitution of Leu for Pro-193 in the insulin receptor in a patient with a genetic form of severe insulin resistance

Mutations have been Identified In the Insulin receptor (IR)gene in patients who are insensitive to Insulin action. We studiedan extremely Insulin resistant patient whose Insulin bindingto Epstein-Barr virus (EBV) transformed lymphocytes was severelyreduced. Transmembrane signalling, evaluated as insulin receptorautophosphorylation, was normal. The patient's IR was Immunoprecipitatednormally by AbP6, a polycional antibody directed to the ßsubunit. However, there was an 50% decrease In the affinityof IR immunoprecipitation by a monoclonal antibody (MA-10) directedagainst the subunlt. These observations suggested that therewas likely to be a mutation In the patient's Insulin receptorthat caused misfolding of the IR subunit. Analysis of genestructure by Southern blotting experiments did not reveal anymajor deletion In the IR gene of the proband. Northern blotanalysis showed a normal level of expression of IR gene. Weapplied denaturing gradlent gel electrophoresis (DGGE) as wellas direct sequence analysis to study the 22 exons of IR geneamplified by polymerase chain reaction (PCR) using the proband'sgenomic DNA as a template. We Identified a new missense mutationsubstituting leucine (CTG) for proline (CCG) in homozygous stateat codon 193 In exon 3. Both parents are heterozygous for theLeu 193 mutation. The Leu¹⁹³ mutation was not detected In anyof 75 normal subjects (150 chromosomes), Indicating that itis not a common sequence variant of the Insulin receptor. Inaddition, during the course of screening the patient's DNA withperpendicular DGGE, we Identified two previously unreportedsilent substitutions in exon 9. We conclude that Leu¹⁹³ mutationis likely to be responsible for causing the patient's disease.     

Quantitative ultrasound assessment of bone

In the last two decades, several non-invasive techniques have been developed to measure bone density at axial and peripheral skeletal sites. The dual-energy X-ray absorptiometry (DXA) technique allows accurate measurement of bone density, but does not provide information about the structural and qualitative features of bone, which play an important role in fracture risk determination. Increasing interest in quantitative ultrasonography (QUS) has recently developed; it may be considered a safe and quite inexpensive diagnostic technique. Ultrasound devices routinely measure two parameters: broadband ultrasound attenuation (BUA) and speed of sound (SOS). Two other parameters, stiffness and index of consistency (QUI), can be derived from BUA and SOS. SOS is influenced by the elasticity of bone as well as by its density. BUA is determined by mechanisms of diffraction, scattering and absorption in the bone, marrow and soft tissue. Absorption predominates in cortical bone and scattering in trabecular bone. BUA is a measure of the approximately linear frequency dependence of ultrasound attenuation. Several QUS devices are now available for clinical use for measuring various parameters at skeletal sites with different contents of trabecular and cortical bone. Standardization of instruments is one of the major limitations of this technique today. Many studies have demonstrated that BUA and SOS, measured at any level, can discriminate normal subjects from osteoporotic patients. Moreover, there is evidence documenting the ability of QUS to predict osteoporotic fracture risk and to give further BMD-independent information on bone. QUS at the heel can now be considered as an alternative technique to identify subjects with a high risk of bone fragility. Further studies are needed for better definition of the role of QUS in clinical practice.     

Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure

BackgroundThe enhancement of circulating endothelial progenitor cells (EPCs) obtained by exercise training can be beneficial to patients with cardiac disease. Changes in the levels and differentiation of CD34^pos/KDR^pos EPCs, as well as the plasma concentration of vascular endothelial growth factor (VEGF) and stromal cell-derived factor (SDF)-1 EPC-mobilizing cytokines, were evaluated in patients with chronic heart failure after 8 weeks of supervised aerobic training (SAT) and 8 weeks of subsequent discontinued SAT (DSAT).Methods and ResultsThe levels of circulating EPC and EPC differentiation potential of 22 patients who underwent SAT were studied by fluorescence-activated cell sorter analysis and colony forming-unit assay, respectively. The plasma levels of VEGF and SDF-1 were measured by enzyme-linked immunosorbent assay. In response to SAT, the levels of both EPC and VEGF/SDF-1 markedly increased (P < .001 vs baseline) but returned to the baseline levels after DSAT. A similar change was observed with the EPC clonogenic potential, but on DSAT the baseline level was incompletely attained.ConclusionsIn response to SAT, patients with chronic heart failure show enhanced EPC levels and clonogenic potential that is mirrored by increased plasma VEGF and SDF-1 levels. DSAT can interfere with the maintenance of training-acquired VEGF/SDF-1-related EPC levels and clonogenic potential.     

Biological qualification of blood units: Considerations about the effects of sample''s handling and storage on stability of nucleic acids

BACKGROUND: In transfusional setting introduction of nucleic amplification technique (NAT) for HBV-DNA, HCV-RNA and HIV-RNA in biological qualification of blood units suggest some problems. At first the opportunity to operate on mini-pool, at second the need to store the samples at +4 degrees C. The authors therefore have tried to estimate the impact of these conditions on the operativity of NAT testing in the transfusional setting. METHODS: The following parameters has been estimated: distribution of viral-load in untreated subjects, stability of nucleic acids during storage at +4 degrees C, stability of nucleic acids after repeated cycles of freezing and defrosting, robustness of the test to the cross-contamination, definition of the detection-limit (95%). Quantitative tests has been performed by using the following kits: Cobas Amplicor HBV Monitor, Cobas Amplicor HCV Monitor, Cobas Amplicor HIV Monitor; the qualitative tests has been performed by using the following kits: Ampliscreen HBV, Ampliscreen HCV 2,0, Ampliscreen HIV 1,5 all supplied by Roche Molecular System (Brancburg, NJ). Results: Viral load in untreated subjects showed wide variation for HBV, HCV and HIV. HBV has been demonstrated much stable to the conservation +4 degrees C also until 168 h while for HCV and HIV a greater decrease of the viral-load was observed. For all and three virus the conservation to +4 degrees C until 72 h does not seem to involve meaningful fall in the viral-load. A remarkable reduction of the viral-load has been observed after five cycles of freezing and defrosting. All the tests showed a good robustness to cross-contamination. The detection-limit (95%) was 8 U/ml for HBV, 21 U/ml for HCV and 27 copy/ml for HIV. CONCLUSIONS: Samples for NAT testing, can be stored until 72 h to +4 degrees C without appreciable lowering of the viral-load. Repeated cycles of changes of state should be avoided. The tests showed a good robustness to cross-contamination. NAT tests for biological qualification of blood units had a minimal sensibility around 50 (copy/unit/ml). In our experience the detection-limit (95%) was 21 U/ml for HCV, 27 copies/ml for HIV, 8 U/ml for HBV. The availability of NAT test for HBV-DNA, HCV-RNA e HIV-RNA, sensitive and reliable, together with epidemiological data, suggest the opportunity to place side by side, in the biological qualification of the blood units, to add the tests for HBV-DNA and HIV-RNA to the test for HCV-RNA mandatory by low, in Italy in the biological qualification of blood units.