Expanding the spectrum of neurological disease associated with Epstein-Barr virus activity

The purpose of this study was to delineate the spectrum of neurological diseases attributed to Epstein-Barr virus (EBV) activity. The approach was a retrospective study on patients with EBV activity proven by a positive EBV antibody-specific index (AI) and/or cerebrospinal fluid (CSF) PCR. One hundred six children and adults (AI positive = 77, AI + PCR positive = 3, PCR positive = 26) were identified, most with reactivated infections. Twenty-eight showed typical EBV-related diseases (encephalitis, neuritis, meningitis), 19 further infections (HSV encephalitis, neuroborreliosis, HIV infection, bacterial meningitis), nine immune-mediated disorders (multiple sclerosis, optic neuritis), and 50 further diseases not typical for EBV. The highest AI values occurred in patients with encephalitis. No relationship between disease category or AI values and viral loads was found. Additional reanalysis of 1,500 consecutive CSF EBV PCR studies revealed the highest positive rates among patients with further infections (n = 18/227, 7.9%) but lower rates among patients with typical EBV-related disorders (5/395; 1.3%), immune-mediated disorders (n = 2/174; 1.1%) and other conditions (n = 4/704; 0.6%). Intrathecal EBV activity is not restricted to typical EBV-related disorders, unexpectedly frequent in further CNS infections and also present in non-inflammatory conditions. Prospective studies should assess the pathogenic role of EBV in these different diseases.     

Thyroid hormones and their correlations with serum glucose, beta hydroxybutyrate, nonesterified fatty acids, cholesterol, and lipoproteins of high-yielding dairy cows at different stages of lactation cycle

The relationships between circulating thyroid hormones and serum glucose, beta hydroxybutyrate (BHB), nonesterified fatty acid (NEFA), cholesterol and lipoproteins of high-yielding dairy cows were studied in 125 adult Holsteins at various stages of lactation cycle: early (far-off) dry period (n = 24), late (close-up) dry period (n = 10), fresh cows (n = 22), early lactation (n = 13), mid-lactation (n = 27) and late lactation (n = 29). Decreased levels of thyroxin (T₄) and triiodothyronine (T₃) were noted in peripartum cows which were extended to mid- and late-lactation cows, respectively. In fresh cows, glucose showed correlations with T₄ (r = 0.619, P < 0.01) and T₃ (r = 0.627, P < 0.01). In the same cows there were correlations between T₄ and BHB (r = 0.590, P < 0.01) and NEFA (r = 0.470, P < 0.01). In late dry cows, free thyroxin (fT₄) showed correlations with triglyceride (TG) and very-low-density lipoprotein (VLDL; r = −0.638, P < 0.05). Pooled data of the various stages of lactation cycle revealed correlations between glucose and T₄ (r = 0.20, P < 0.05), glucose and T₃ (r = 0.395, P < 0.01), cholesterol and T₃ (r = −0.201, P < 0.05), and free triiodothyronine (fT₃) and high-density lipoprotein (HDL; r = 0.178, P < 0.05). It appears that the time and the pattern of changes and/or correlations of serum thyroid hormones and lipid fractions may vary among dairy cows and other animals.     

Increased Extracellular Survivin in the Synovial Fluid of Rheumatoid Arthritis Patients: Fibroblast-like Synoviocytes as a Potential Source of Extracellular Survivin

Survivin belongs to the family of inhibitor of apoptosis proteins and plays an important role in the hyperplastic growth of tissues and tumors. In this study, we assessed the expression of survivin in rheumatoid synovial fluids (SF) and synovial tissues (ST) of rheumatoid arthritis (RA) patients in order to investigate the role of extracellular survivin in the pathogenesis of RA. The survivin level from SF was significantly higher in RA patients (n = 38) than in osteoarthritis patients (n = 18; 10.68 ± 2.76 vs. 1.0 ± 0.56 pg/ml, p = 0.02). In addition, SF survivin level was higher in erosive RA patients (n = 23) than in non-erosive RA patients (n = 15; 15.26 ± 4.26 vs. 4.47 ± 1.12 pg/ml, p = 0.05). SF survivin level in RA was positively correlated with disease activity score 28, but did not reach statistical significance (r = 0.309, p = 0.07). RA SF survivin level was also positively correlated with peripheral blood leukocyte counts (r = 0.443, p = 0.005). The immunohistochemical staining and Western blot analysis revealed survivin expression in the ST and fibroblast-like synoviocytes of RA patients, respectively. These findings suggest that extracellular survivin may be produced from rheumatoid FLS and may play an important role in the destructive RA process.     

Association between serum cortisol and progesterone concentrations and the infiltration of immune cells in the endometrium of gilts with vaginal discharge

The objective of the study was to investigate an association between serum cortisol and progesterone (P₄) concentrations and the distribution of immune cells in the endometrium of the gilts with vaginal discharge. Genital organs from 39 Landrace×Yorkshire crossbred gilts culled owing to vaginal discharge problem were collected from two commercial swine herds in Thailand. The estrous stage and gross pathology were examined. Blood samples were collected from the jugular vein prior to being slaughtered. Serum P₄ and cortisol were analyzed by means of enzyme immunoassay. The samples observed were in inactive (n = 4), follicular (n = 10), and luteal (n = 25) phases. They, afterwards, were processed in hematoxylin and eosin sections. The endometrium of the gilts was histologically divided into three layers, i.e., epithelial, subepithelial connective tissue, and glandular connective tissue layers. Immune cells, i.e., lymphocytes, neutrophils, eosinophils, macrophages, and plasma cells, in each layer were quantified under a light microscope (×400). The results revealed that mean serum cortisol was 430.6 ± 68.3 nmol/l. Serum P₄ varied by ovarian status. Serum P₄ of the gilts in the luteal phase was higher than those in the follicular phase (88.3 ± 7.7 versus 20.6 ± 6.2 nmol/l, P < 0.05). As for the endometrium condition, the gilts were classified into acute/subacute endometritis (n = 13), chronic endometritis (n = 9), and normal endometrium (n = 17). Neutrophils were the main local immune cells in the epithelial layer. Lymphocytes were the dominant population in the subepithelial and glandular connective tissue layers. Generally, the serum cortisol tended to negatively correlate with lymphocytes in the subepithelial connective tissue layer (r = −0.28, P = 0.081). In the gilts with acute/subacute endometritis, no correlation among serum cortisol, P₄, and immune cells was observed. In chronic endometritis gilts, only a negative correlation was remarked between P₄ and epithelial lymphocytes (r = −0.83, P = 0.010), epithelial neutrophils (r = −0.79, P = 0.019), and subepithelial neutrophils (r = −0.73, P = 0.025). In the gilts with normal endometrium, P₄ negatively correlated with subepithelial neutrophils (r = −0.55, P = 0.022) while positively correlated with subepithelial macrophages (r = 0.54, P = 0.024) and subepithelial eosinophils (r = 0.60, P = 0.011).     

Evaluation of the Lactate Pro blood lactate analyser

An evaluation of the hand-held portable Lactate Pro Analyser (KDK) was undertaken to assess its accuracy, reliability and versatility. Capillary blood samples were drawn from elite athletes in both laboratory and field settings and analysed in parallel. Accuracy was determined in relation to three other lactate analysers: (1) the ABL 700 Series Acid-Base analyser (n=172 cases), (2) the Accusport Lactate Meter (n=118 cases), and (3) the YSI 2300 Stat lactate analyser (n=22 cases). The level of agreement was determined over the range of 1–18 mM. The repeatability of results between two different Lactate Pro analysers was also determined over the same range. Versatility was assessed in the field, where the Lactate Pro was used with elite athletes under a range of outdoor and indoor testing conditions. The correlations between the Lactate Pro and the ABL 700 Series Acid-Base analyser, YSI 2300 and Accusport were r=0.98, r=0.99, r=0.97. The correlation between the two Lactate Pro analysers on the same sample (n=96 cases) was r=0.99. The level of agreement between the Lactate Pro and other analysers was generally less than ±2.0 mM over the physiological range of 1.0–18.0 mM (range of mean difference: −0.06 mM to 0.52 mM). The Lactate Pro was easy to operate and successfully completed the sample analysis in 100% of the tests performed. In summary, the Lactate Pro is accurate, reliable and exhibits a high degree of agreement with other lactate analysers. Accepted: 18 October 1999     

Peripheral and brachial blood pressure during standardized occupation-related tests in normotensive and mild hypertensive men and women

We investigated the usefulness of peripheral blood pressure (BP) measurement in the assessment of strain in occupational physiology. Our hypothesis was that the brachial and peripheral BP reflect physiologically different events under various occupation-related demands in normotensive (NT) and hypertensive (HT) people. A group of 20 female and 20 male subjects with unmedicated mild hypertension that had been diagnosed by ambulatory blood pressure monitoring [awake time systolic/diastolic BP (BP_s/BP_d) 142.9 (SD 11.3)/86.4 (SD 6.2) mmHg] and 40 NT matched by age and sex [BP_s/BP_d 120.0 (SD 9.8)/75.6 (SD 5.9) mmHg] attended a laboratory session to undertake mental arithmetic tasks, a fingergrip test and submaximal cycle ergometry. Brachial and peripheral BP as well as heart rate were measured using a sphygmomanometer and an continuously automatic blood pressure measuring device on the finger, respectively. The peripheral BP_s was higher than brachial BP_s, BP_d was similar for peripheral and brachial BP except during cycle ergometry. Associations between the levels of brachial and peripheral BP depended on demands and did not explain more than 42% of the common variance. The highest correlations between the two BP methods were observed during habituation, recovery and mental demands, and weak correlations during cycle ergometry. For peripheral BP_s and BP_d we found significant correlations in all phases of the test (r=0.58 to 0.86, P < 0.001), also in ergometry (NT r=0.62, P < 0.001, HT r=0.53, P < 0.001), in contrast to the brachial BP. Peripheral BP differentiated the two BP groups (57.5%–72.5% correctly classified) which had been grouped by daily measurement of brachial BP, but brachial BP was superior in this respect with 65.0%–87.5% being correctly classified. These results supported the suggestion that the combined measurement of peripheral and brachial BP provides complementary information regarding physiological changes in NT and HT in different situations. Accepted: 30 August 1999     

Subacute Inflammatory Activation in Subjects with Acute Coronary Syndrome and Left Ventricular Dysfunction

Several lines of evidence indicate that increased inflammatory cytokine levels can be used for risk prediction in patients with acute coronary syndrome (ACS). This study therefore aimed to evaluate correlations between levels of soluble interleukin (IL)-2 receptor (sIL-2r), IL-6, and IL-8 and in-hospital incidence of acute heart failure (AHF) and left ventricular (LV) systolic dysfunction in the subacute phase of ACS. In 48 consecutive patients with ACS, circulating levels of sIL-2r, IL-6, and IL-8 were ascertained 72–96 h after onset of symptoms. Clinical data, LV function, and in-hospital incidence of AHF were also evaluated. IL-8 levels were significantly higher in patients with pulmonary edema (1,829 ± 2,496 vs 456 ± 624 pg/ml, p < 0.05); sIL-2r, IL-6, and IL-8 levels were increased proportionally to Killip class (r = 0.35, p < 0.05; r = 0.48, r = 0.47, p < 0.01) and in patients with LV ejection fraction (LVEF) < 30%. Levels of sIL-2r were inversely related to LVEF in subjects with acute myocardial infarction (r = −0.51, p < 0.05). Soluble IL-2r and IL-8 levels were related to mitral regurgitation severity (r = 0.34, p < 0.05; r = 0.37, p < 0.05). Levels of sIL-2 were proportional to LV end-diastolic diameter (r = 0.49, p < 0.001) and LV end-systolic diameter (r = 0.58, p < 0.001). Number of cytokines with circulating values above upper level of normal was significantly correlated with Killip class and LVEF (r = 0.40, r = −0.38, p < 0.05). sIL-2r, IL-6, and IL-8 are increased in patients with ACS and systolic dysfunction or AHF. These data suggest that inflammatory cytokine activity detectable in peripheral blood may be useful in identifying subjects with a worse clinical course.     

Diagnostic value of procalcitonin in pleural effusions

This study was to determine the diagnostic value of procalcitonin (PCT) in the differentiation of infectious and non-infectious causes of pleural effusion. From January 2005 to April 2005, we measured the PCT levels of pleural effusion from 76 patients using an immunoluminometric assay. The types of pleural infusions studied were para-pneumonic effusion (n = 26), empyema (n = 7), tuberculous pleurisy (n = 8), malignant pleural effusion (n = 25) and transudative pleural effusion (n = 8). The PCT levels were low in transudative pleural effusions (0.188 ± 0.077 ng/mL) and tuberculous pleurisy (0.130 ± 0.069 ng/mL), but high in empyema (5.147 ± 3.056 ng/mL), para-pneumonic effusion (1.091 ± 0.355 ng/mL), and malignant pleural effusion (0.241 ± 0.071 ng/mL). The receiver-operating characteristic curve analysis for an optimal discrimination between empyema and para-pneumonic effusion from non-para-pneumonic effusion could be performed at a cut-off point of 0.18 ng/mL with area under the curve of 0.776 (sensitivity: 69.7%, specificity: 72.1%). The correlation was found between pleural effusion PCT and serum PCT levels in 16 patients (r ² = 0.967, p < 0.001). In conclusion, a high pleural effusion PCT level suggests the presence of empyema and para-pneumonic effusion.     

Pathology of testis and epididymis in native goats in southern Iran

A slaughterhouse-based survey was conducted to determine the type and the prevalence of lesions in the testis and epididymis of native bucks reared in southern Iran. Testis, epididymis, and tunica, which belonged to 425 bucks of various age groups, were inspected. The specimens were collected randomly during a 6-month period. Various abnormalities in testis and the epididymis were observed. Grossly, testicular mineralization was the most prevalent abnormality (n = 183, 45%) followed by degeneration or hypoplasia (n = 26, 6.4%), adhesion (n = 20, 4.9%), cryptorchidism (n = 12, 2.9%), congenital testicular cyst (n = 9, 2.2%), abscess (n = 4, 0.9%), and orchitis (n = 1, 0.2%). As the age of the bucks increased, the percentages of mineralization increased significantly (p < 0.05). Based on the results of the gross examination, congenital epididymal cysts were the most prevalent abnormality (n = 57, 14.4%) then followed by epididymal abscess (n = 12, 2.9%), melanosis (n = 10, 2.5%), and epididymitis (n = 3, 0.7%). Congenital epididymal cysts, 1 to 2 mm in diameter, were mostly located on the head of the epididymis. On histopathological examination, mineralization showed the highest prevalence rate in testis followed by hypoplasia and degeneration, besnoitiosis, orchitis, and edema. Besnoitiosis was also the predominant lesion in the head and tail of the epididymis followed by epididymitis, hypoplasia or degeneration, melanosis, and sperm granuloma. Besnoitia cysts were found in 11.3% of the testes, 14.1% of the epididymal heads, and 7.5% of the epididymal tails.     

Safety and tolerability of sputum induction in adolescents and adults with suspected pulmonary tuberculosis

Sputum induction by the inhalation of hypertonic saline may increase the yield of microbiological diagnosis of pulmonary tuberculosis (TB). This is particularly relevant in paucibacillary TB, such as in children or human immunodeficiency virus (HIV)-infected patients. Sputum induction must be shown to be safe and tolerable in community settings where invasive diagnostic methods are unavailable. The objective of this study was to describe the changes in physiological parameters and adverse events occurring during sputum induction in ambulatory adult and adolescent TB suspects recruited in community clinics. Sputum induction was performed in HIV-infected (n = 35) and HIV-uninfected (n = 67) TB suspects (n = 102). Oxygen saturation (%), blood pressure (mm Hg), heart rate (/minute), respiratory rate (/minute), and adverse events were monitored at baseline, continuously during the salbutamol pre-treatment and saline nebulization phases, and for 30 min afterwards. During nebulization, there was a statistically significant increase in oxygen saturation (1%, p < 0.0001), systolic BP (7 mm Hg, p < 0.0001), and diastolic BP (2 mm Hg, p = 0.008). Post-nebulization decrease in the systolic BP occurred (4 mm Hg, p = 0.016). These changes were not considered to be clinically significant. Eight minor, transitory, self-resolving adverse events occurred (labored breathing, n = 2; chest pain, n = 2; paroxysmal coughing, n = 1; elevated heart rate, n = 1; vomiting, n = 1; hypotension, n = 1), leading to procedure termination in four participants. No serious adverse events occurred. Induced sputum is safe, tolerable, and feasible in adult and adolescent TB suspects in a community healthcare setting.     

Design considerations in the development and application of microdisc electrode arrays (MDEAs) for implantable biosensors

The use of microlithographically fabricated Microdisc Electrode Arrays (MDEAs) in the development of implantable voltammetric biosensors necessitates design criteria that balances the overall footprint of the device with the advantages to be derived from large separation distances between non-interacting microdisc elements. Using the dynamic electroanalytical techniques of Multiple Scan Rate Cyclic Voltammetry (MSRCV) experiments with finite element simulations and Electrochemical Impedance Spectroscopy with equivalent circuit modeling, three unique MDEA designs; MDEA 050 (r = 25 μm, 5,184 discs), MDEA 100 (r = 50 μm, 1,296 discs) and MDEA 250 (r = 125 μm, 207 discs) of constant critical dimensions (center-to-center d/r = 4) and area (A = 0.1 cm²) were studied in 1.0 mM ferrocene monocarboxylic acid (FcCO₂H) solution (in 0.1 M Tris/0.1 M KCl buffer, pH = 7.2). The critical disc-to-disc spacing (d/r) required to archive 67% of maximal current response was defined as optimal. Based on the predictive model, new MDEA designs; MDEA 001 (r = 0.5 μm, 127,324 discs), MDEA 002.5 (r = 1.25 μm, 20,372 discs), MDEA 005 (r = 2.5 μm, 5,093 discs), MDEA 010 (r = 5 μm, 1,273 discs), MDEA 015 (r = 7.5 μm, 566 discs), MDEA 020 (r = 10 μm, 318 discs) were simulated at 10 and 100 mV/s. The final disc count of each MDEA was dictated by the need to maintain a comparable electroactive area between the MDEAs, which was chosen to be 0.001 cm², which in turn was dictated by the need to generate sufficient electrochemical current to be comfortably measured by common electrochemical detectors.     

A Retrospective Study of Feline Pancytopenia

To better define the incidence and causes of feline pancytopenia we reviewed cases of pancytopenia submitted to the University of Minnesota, Veterinary Teaching Hospital over an 18-month period. Pancytopenia was defined as a combination of anaemia (packed cell volume < 26%), neutropenia (segmented neutrophils < 3000/μl), and thrombocytopenia (platelet counts < 200 000/μl). Of 2011 complete blood counts reviewed, pancytopenia was detected in 56 (2.8%). Associated clinical disorders included drug-associated disorders (n = 9) infectious diseases (n= 14) immune-mediated haematological diseases (n= 3), chronic renal failure (n = 3), idiopathic aplastic anaemia (n= 3), and other causes (n= 5). Results of this study indicate that feline pancytopenia has multiple causes and that the prognosis is dependent on the cause of the pancytopenia. Differentiation of the various causes of pancytopenia requires a systematic approach that includes evaluation of infectious, drug-induced, and immune-mediated causes, and examination of bone marrow.     

A Retrospective Study of Feline Pancytopenia

To better define the incidence and causes of feline pancytopenia we reviewed cases of pancytopenia submitted to the University of Minnesota, Veterinary Teaching Hospital over an 18-month period. Pancytopenia was defined as a combination of anaemia (packed cell volume < 26%), neutropenia (segmented neutrophils < 3000/μl), and thrombocytopenia (platelet counts < 200 000/μl). Of 2011 complete blood counts reviewed, pancytopenia was detected in 56 (2.8%). Associated clinical disorders included drug-associated disorders (n = 9) infectious diseases (n= 14) immune-mediated haematological diseases (n= 3), chronic renal failure (n = 3), idiopathic aplastic anaemia (n= 3), and other causes (n= 5). Results of this study indicate that feline pancytopenia has multiple causes and that the prognosis is dependent on the cause of the pancytopenia. Differentiation of the various causes of pancytopenia requires a systematic approach that includes evaluation of infectious, drug-induced, and immune-mediated causes, and examination of bone marrow.     

Treatment of Exacerbations of Multiple Sclerosis without the Use of Corticosteroids: The Role of Metabolic and Antioxidant Therapy

A multicenter randomized post-registration case-control study was performed with 94 patients with relapsing-remitting and secondary progressive multiple sclerosis (MS) in exacerbation. The patients were divided into two groups: group 1 (n = 53) received Cytoflavin and basal therapy (Trental and group B vitamins), while group 2 (n = 41) received only basal therapy. After five days, treatment results were used to further divide each group into two subgroups: patients of subgroup 1A (n = 22), with clear positive treatment effects, continued to receive Cytoflavin and basal therapy; group 1B (n = 31) additionally received corticosteroid (methipred) pulse therapy; subgroup 2A (n = 14), with significant positive effects, continued to receive basal therapy; group 2B (n = 27) additionally received corticosteroid pulse therapy. The complex treatment, with Cytoflavin, Trental, group B vitamins, and corticosteroids, was found to be safe and well tolerated. The positive effects of including Cytoflavin in complex treatment consisted of a reduction in the need for corticosteroids: 41.5% of patients receiving Cytoflavin no longer needed them, compared with 34% of patients receiving only basal therapy. Patients given Cytoflavin showed more significant regression of neurological symptomatology on the EDSS than the group not given Cytoflavin; there were no patients with no treatment response; lipid peroxidation and anti-myelin basic protein antibody levels decreased; cognitive functions improved.     

Performance of two commercial blood IFN-γ release assays for the detection of Mycobacterium tuberculosis infection in patient candidates for anti-TNF-α treatment

The reactivation of latent tuberculosis (TB) is a major complication of tumor necrosis factor (TNF)-α inhibitors. Screening for TB infection is recommended before anti-TNF therapy is initiated; however, the use of tuberculin skin testing (TST) is controversial, due to the high rate of false-negative results in patients receiving immunosuppressive treatment. To compare the performance of two commercial interferon (IFN)-γ release assays (IGRA), T-SPOT.TB (TS-TB) and QuantiFERON-TB Gold “In-tube” (QFT-GIT), with TST for the detection of TB infection in patients due to start anti-TNF therapy, 69 human immunodeficiency virus (HIV)-negative Italian patients (mean age: 45.2 ± 12.6 years; male=39) were enrolled between September 2005 to August 2006. Patients affected by rheumatoid arthritis (n = 18), psoriatic arthritis (n = 26), ulcerous rectocolitis (n = 6), and Crohn’s disease (n = 19) were tested simultaneously with TST, TS-TB, and QFT-GIT. Overall, 26% of patients were positive by TST, 30.4% by TS-TB, and 31.8% by QFT-GIT. Agreement with TST was similar (κ = 0.21, p = 0.0002 and κ = 0.26, p < 0.001, respectively). In 11 TST-negative cases, IFN-γ release assays were positive. In addition, in seven Mantoux-positive cases with no TB risk factors, TST result agreement was achieved with at least one blood test. Indeterminate results were detected in 5.8% and 2.8% of cases, respectively, with TS-TB and with QFT-GIT (p = not significant [ns]). In conclusion, our results suggest that IGRAs may be helpful for screening purposes in patient candidates for anti-TNF therapy to confirm positive TST results and in selected cases when false-negative results are suspected. The utility of blood tests in patients with low or no TB risk remains to be assessed.     

Prevalence of toxin genes in consecutive clinical isolates of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and clinical impact

Even if Panton–Valentine leukocidin (PVL), toxic shock syndrome toxin-1 (TSST-1), staphylococcal enterotoxins (SEB and SEC), and exfoliative toxins (ETA and ETB) may be associated with severe infections, the clinical significance of their presence in clinical isolates of Staphylococcus aureus remains poorly documented. In this study, we evaluated the prevalence of toxin genes and the relationship between their presence and the severity of infection. We screened for the presence of these six toxin genes among 186 consecutive S. aureus clinical isolates (resistant or not to methicillin) during a two-month period. We compared the toxin gene profile between strains recovered from patients presenting uncomplicated infections (n = 151) and from patients suffering from severe infections (n = 35). At least one toxin gene was detected in 55 (29.6%) isolates as follows: pvl (n = 1), tst + sec (n = 5), seb (n = 19), seb + sec (n = 1), sec (n = 28), and eta (n = 1). The proportion of toxin-producing strains among patients with uncomplicated infections (27.8%) and patients with severe infections (37.1%) was not statistically different (p = 0.3044), even if the severity of infection tended to be associated with the presence of sec (p = 0.0655). Although the prevalence of toxin genes was relatively high herein, no statistically significant association between the severity of infection and the presence of toxin genes was observed.     

Cyclin D3 gene amplification in bladder carcinoma in situ

Carcinoma in situ (CIS) is a non-papillary high-grade, potentially aggressive, and unpredictable manifestation of bladder urothelial carcinoma. The aim of this study was to assess patterns of Cyclin D3 gene amplification in Bacillus Calmette-Guerin (BCG)-treated CIS and correlate gene status with recurrence-free and progression-free survival. A sequential cohort series of 28 primary (isolated) or secondary (concomitant) bladder CIS samples in which there was enough tissue material to assess Cyclin D3 gene status by fluorescent in situ hybridization was the study group. Cyclin D3 gene amplification was present in 29% of secondary CIS; none of primary CIS samples had Cyclin D3 gene amplification. Cyclin D3 amplification was related to recurrence- (p = 0.046) and progression-free survival (p = 0.002). Type of bladder CIS (primary vs. secondary) was unrelated to recurrence- or progression-free survival in the current series. Cox’s regression analysis selected Cyclin D3 as an independent predictor of progression-free survival (p = 0.041, relative risk = 61.503, 95% confidence interval = 1.1–274.710). None of primary CIS cases recurred on follow-up; nine secondary CIS recurred and four of them progressed to invasive bladder carcinoma HG T1 (n = 1), T2b N0M0 (n = 1), T3b N1M0 (n = 1) and T4aN1M1 (n = 1). Mean recurrence ± SD (months) occurred at 19.5 ± 2.06 (95% (confidence interval (CI)), 15.5–23.6); mean progression (months) occurred at 23.8 ± 1.46 (95% (CI), 20.9–26.7). Our study suggests that Cyclin D3 gene amplification might be a predictor of aggressiveness in BCG-treated CIS.     

Comparative study on determination of plasma thyroid hormones by chemiluminescence and electrochemiluminescence immunoassay methods in sheep

The aim of this study was to compare plasma thyroid hormone concentrations by both chemiluminescence (CLIA) and electrochemiluminescence immunoassay (ECLIA) methods in sheep. Blood samples were taken from the jugular vein of 25 clinically healthy, non-pregnant adult ewes. The plasma was analyzed to determine thyroxine (T₄), tri-iodothyronine (T₃), free thyroxine (fT₄) and free tri-iodothyronine (fT₃) concentrations. The data from this study indicates, there were significant differences in the T₄ (P < 0.0001), T₃(P < 0.01) and fT₃(P < 0.01) concentrations between the two methods, and the levels of these hormones were higher when using the ECLIA method. In determining thyroid hormones using the CLIA and ECLIA methods, significant positive correlations were found between T₄ and fT₄ (P < 0.001, r = 0.703; P < 0.0001, r = 0.806) also between T₃ and fT₃ (P < 0.0001, r = 0.922; P < 0.0001, r = 0.923) concentrations, respectively. The linear regression analysis of these hormones showed that the CLIA and ECLIA results were significantly correlated (T₄ (P < 0.01, r = 0.583), T₃ (P < 0.0001, r = 0.898), fT4 (P < 0.0001, r = 0.796) and fT₃ (P < 0.0001, r = 0.898).     

Frequent concomitant inactivation of <Emphasis Type="Italic">miR-34a</Emphasis> and <Emphasis Type="Italic">miR-34b/c</Emphasis> by CpG methylation in colorectal,pancreatic, mammary,ovarian, urothelial,and renal cell carcinomas and soft tissue sarcomas

The microRNA encoding genes miR-34a and miR-34b/c represent direct p53 target genes and possess tumor suppressive properties as they mediate apoptosis, cell cycle arrest, and senescence. We previously reported that the miR-34a gene is subject to epigenetic inactivation by CpG methylation of its promoter region in primary prostate cancer and melanomas, and in 110 different cancer cell lines of diverse origin. Here we analyzed the methylation status of miR-34a and miR-34b/c in additional primary tumors of divergent sites. We found methylation of miR-34a or miR-34b/c in formalin-fixed, paraffin-embedded (FFPE) tumor samples from 178 patients with the following frequencies: colorectal cancer (74% miR-34a, 99% miR-34b/c; n = 114), pancreatic cancer (64%, 100%; n = 11), mammary cancer (60%, 90%; n = 10), ovarian cancer (62%, 69%; n = 13), urothelial cancer (71%, 57%; n = 7), and renal cell cancer (58%, 100%; n = 12). Furthermore, soft tissue sarcomas showed methylation of miR-34 gene promoters in FFPE samples (64%, 45%; n = 11), in explanted, cultured cells (53%, 40%; n = 40), and in frozen tissue samples (75%, 75%, n = 8). In the colorectal cancer samples a statistically significant correlation of miR-34a methylation and the absence of p53 mutation was detected. With the exception of sarcoma cell lines, the inactivation of miR-34a and miR-34b/c was concomitant in most cases. These results show that miR-34 inactivation is a common event in tumor formation, and suggest that CpG methylation of miR-34a and miR-34-b/c may have diagnostic value. The mutual exclusiveness of miR-34a methylation and p53 mutation indicates that miR-34a inactivation may substitute for loss of p53 function in cancer.