S100B as a new fecal biomarker of inflammatory bowel diseases.

OBJECTIVE: S100 proteins are demonstrated to exert a protective role in the gastrointestinal tract. In the present study, we investigated whether S100B protein, that is typically expressed by enteroglial cells, is detectable in feces and could be a useful noninvasive indicator of gut chronic inflammation. PATIENTS AND METHODS: This clinical prospective study included n=48 patients suffering Crohn's disease (CD) or ulcerative colitis (UC) and non IBD-controls. The clinical disease activity was evaluated using Harvey-Bradshaw or Mayo Score Index while the diagnosis of IBD was defined based on standard endoscopic and histological criteria. S100B and calprotectin were extracted and analyzed using commercial enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: Unlike calprotectin, S100B was significantly decreased in both CD and UC compared to non IBD-patients. The strongest quantitative alterations of S100B were detected concomitantly with signs of active or quiescent disease, including high/normal expression of fecal calprotectin, mucosal damage/cryptitis, mucin depletion and inflammatory infiltrate, as defined by endoscopic evaluation and histological analysis. At the onset of disease and under no Infliximab-based therapy, the lowest was detected suggesting that S100B in feces could have a potential diagnostic value for IBD. CONCLUSIONS: Testing for S100B and calprotectin could be a useful screening tool to better predict IBD activity.

Impact of HIV-1 Infection and Antiretroviral Therapy on Bone Homeostasis and Mineral Density in Vertically Infected Patients.

Daily assumption of antiretroviral drugs and HIV-related immune activation lead to important side effects, which are particularly evident in vertically infected patients. Bone homeostasis impairment and reduction of bone mineral density (BMD) is one of the most important side effects. Primary aim of this study is to assess the prevalence of bone homeostasis alterations in a group of vertically infected patients; secondary aim is to analyze the relationship between bone homeostasis alterations and anthropometric data, severity of HIV infection, and antiretroviral therapy. We studied 67 patients with vertically transmitted HIV-1 (aged 6-31 years), followed by the Pediatric Infectious Disease Unit of the University Hospital of Padua, Italy. We analyzed bone turnover markers (P1NP and CTx) and we performed lumbar spine and femoral dual energy X-ray absorption densitometry (DXA). Personal and anthropometric data and information on HIV-infection severity and antiretroviral therapy were collected for all patients. We found that BMD values recorded by DXA showed a significant correlation with age, race, BMI, physical activity, and antiretroviral therapy duration. P1NP was increased in 43% of patients, while CTX in 61% of them. P1NP alteration was related to age, race, BMI, physical activity, therapy duration, and ever use of protease inhibitors and nucleotide reverse transcriptase inhibitors. CTX alteration was found to be correlated only with age. In conclusion, our study confirms that a wide percentage of HIV vertically infected patients show reduced BMD and impaired bone homeostasis. Strict monitoring is needed in order to early identify and treat these conditions.

Quality indicators to detect pre-analytical errors in laboratory testing.

The identification of reliable quality indicators (QIs) is a crucial step in enabling users to quantify the quality of laboratory services. The current lack of attention to extra-laboratory factors is in stark contrast to the body of evidence pointing to the multitude of errors that continue to occur, particularly in the pre-analytical phase. The ISO 15189: 2012 standard for laboratory accreditation defines the pre-analytical phase, and recognizes the need to evaluate, monitor and improve all the procedures and processes in the initial phase of the testing cycle, including those performed in the phase of requesting tests and collecting samples, the so-called "pre-pre-analytical phase". Therefore, QIs should allow the identification of errors and non-conformities that can occur in all steps of the pre-analytical phase. Traditionally, pre-analytical errors are grouped into identification and sample problems. However, appropriate test requesting and complete request forms are now recognized as fundamental components in providing valuable laboratory services. The model of QIs developed by the Working Group of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) includes indicators related to both identification and sample problems as well as all other pre-analytical defects, including those in test requesting and request forms. It, moreover, provides the framework (with objective criteria) necessary for promoting the harmonization of available QIs in the pre-analytical phase.

An integrated system for monitoring the quality of sample transportation.

OBJECTIVES: Due to the consolidation of laboratory testing facilities, there is an increasing need for systems able to assure quality and safety in biological sample transportation, although little evidence on this aspect is available in literature. DESIGN AND METHODS: An integrated system for sample transportation, implemented and monitored over a five-year period by our team, consists of secondary and tertiary containers, a device for temperature and time recording, and a system manager allowing the acceptance or rejection of biological samples through the immediate visualization and validation of registered data. RESULTS: Data collected between 2009 and October 2011, after a preliminary phase for optimizing the temperature inside the containers, demonstrated the frequency of transportations at an acceptable temperature (<20 °C) had increased and that of transportations at an excessively high temperature (>25 °C) had decreased by ~80%. CONCLUSIONS: The integrated system and related operating instructions allow improvement in the quality of sample transportation over time.

Uterine artery blood flow volume in pregnant women with an abnormal pulsatility index of the uterine arteries delivering normal or intrauterine growth restricted newborns.

The aim of this study was to assess and compare uterine artery (UtA) blood flow volume in pregnant patients with an abnormal uterine Doppler pulsatility index (PI) who delivered fetuses with an appropriate weight for gestational age (AGA) or with intrauterine growth restricted (IUGR). We prospectively recruited singleton pregnancies with abnormal uterine arteries P.I. between 18 and 38 weeks of gestation regardless of estimated fetal weight (EFW). Vessel diameter and blood flow velocity were measured along the UtA upstream to the vessel bifurcation in both the right and left UtAs. Uterine blood flow volumes measured in these pregnancies were compared to historical Control-pregnancies. Forty-three patients delivered at term a normal weight newborn (AGA-pregnancies). Thirty patients delivered growth restricted newborns at 32 weeks (i.r. 29-36w) with a median weight of 1160 gr (i.r. 1000-2065 gr) (IUGR-pregnancies). At mid-gestation (18 + 0 - 25 + 6 weeks + days of gestation) a significantly lower uterine blood flow volume per unit weight was observed between the two study groups and compared to controls: 142 ml/min/kg in IUGR-pregnancies, 217 ml/min/kg in AGA-pregnancies and 538 ml/min/kg in Control-pregnancies. These striking differences in blood flow volume were already present at mid-gestation, at a time when EFW was still normal. In late gestation (27 + 0 - 37 + 6 weeks + days of gestation), pregnancies with an abnormal uterine P.I. showed persistently low UtA flow (<50% of controls) even when corrected for fetal weight: 81 ml/min/kg in IUGR-pregnancies, 105 ml/min/kg in AGA-pregnancies, and 193 ml/min/kg in Control-pregnancies; p < 0.0001. Our findings are consistent with other recent studies regarding the association between reduced uterine blood flow volume and fetal growth restriction. However, the study brings new insight into the finding of abnormal uterine P.I. in normally grown fetuses typically dismissed as "falsely abnormal" or "false positive" findings. Our study suggests that blood flow volume measurement may serve as a new tool to assess this group of patients and possibly those with ischemic placental diseases that may provide some basis for therapeutic interventions.

Blood flow volume of uterine arteries in human pregnancies determined using 3D and bi-dimensional imaging, angio-Doppler, and fluid-dynamic modeling.

The primary aim of this pilot study was to study uterine artery (UtA) blood flow volume in uneventful human pregnancies delivered at term, at mid and late gestation by means of 3D and bi-dimensional ultrasound imaging with angio-Doppler combined with fluid-dynamic modeling. Secondary aims were to correlate flow volume to placental site and to UtA Pulsatility Index (PI). Women with singleton, low-risk pregnancies were examined at mid and late gestation. The structure and course of the uterine artery (UtA) was studied in each patient by means of 3D-angio-Doppler and included vessel diameter D, blood flow velocity and PI (measured along the UtA). Fetal weight estimation and placental insertion site were assessed by ultrasound. A robust fluid-dynamic modeling was applied to calculate absolute flow and flow per unit fetal weight. Mean UtA diameter and blood flow velocity increased significantly (p < 0.0001) from mid-gestation to late gestation from 2.6 mm and 67.5 cm/s, to 3.0 mm and 85.3 cm/s, respectively, yielding an increasing absolute flow troughout gestation. h coefficient, derived by fluid-dynamic modeling to calculate mean velocity, increased significantly from 0.52 at mid-gestation to 0.57 at late gestation. UtA blood flow volume ml/min/kg-fetal weight was significantly higher at mid-gestation than at late gestation (535 ml/min/kg vs 193 ml/min/kg; p < 0.0001). In cases with strictly lateral placentas the ipsilateral UtA accommodates at mid and late gestation 63% and 67% of the total UtA flow. In central placentas UtA flow was evenly distributed between the two vessels. An inverse correlation was observed between PI and blood flow volume ml/min/kg (Pearson's coefficient r = -0.54). Our work confirms the technological and methodological limitations in the measurement of uterine artery blood flow. However, Doppler measurements supported by three-dimensional angio imaging of the uterine vessel, high resolution imaging and diameter measurement, and a robust mathematical model of local circulation adds a genuine new area of investigation into human uterine circulation during pregnancy.

Heat-induced transcription of diphtheria toxin A or its variants, CRM176 and CRM197: implications for pancreatic cancer gene therapy.

Vectors combining the heat shock proteins (HSPs) promoter with the catalytic subunit A of the diphtheria toxin (DTA) or its variants, cross-reacting material (CRM) 176 and 197, were engineered to investigate the effect of bacterial toxins on pancreatic cancer (PC) cells. Three heat-inducible enhanced green fluorescent protein (eGFP)-expression vectors were obtained: V1 (91% homology to HSPA6), V2 (five heat shock elements upstream the minimal HSPA6 promoter) and V3 (V1 and V2 combined). The highest eGFP transcription and translation levels were found in V3 transfected PC cells. The V3 promoter was used to control DTA, CRM176 and CRM197 expression, treatment response being investigated in four PC cell lines. DTAwt or CRM176 transfected cell growth was completely arrested after heat shock. CRM197 toxin presumed to be inactive, caused mild distress at 37 degrees C and induced a 25-50% reduction in cell growth after heat shock. Preliminary in vivo findings showed that heat treatment arrests tumor growth in DTA197 stably transfected PSN1 cells. In conclusion, the efficient HSP promoter identified in this study may be extremely useful in controlling the transcription of toxins such as CRM197, which have lethal dose-related effects, and may thus be a promising tool in PC gene therapy in vivo.

Hypophyseal triplication: case report and embryologic considerations.

SUMMARY: Hypophyseal triplication is malformation that has not been described previously. We present a child with midline abnormalities who underwent epignathus excision at birth. Brain MR imaging revealed 2 paired lateral pituitary glands and an oval midline gland, each with an independent stalk, connected to a thickened third ventricle floor. Because malformations represent a failure in embryogenesis, this case may provide interesting clues on the normal development of the hypophysis.

Risk factors for cerebral glioma in adults: a case-control study in an Italian population.

A case-control study on risk factors for cerebral tumors was conducted on an adult Italian population by the four Neurosurgical Departments of the Veneto Region, i.e. Padua, Treviso, Verona and Vicenza. The study recruited 195 cases of histologically-confirmed cerebral glioma. One hospital control was selected for each case. Cases and controls were matched for age, sex, data of hospitalization and residence. Information on both cases and controls was obtained from a relative. Uninvolved interviewers administered a structured questionnaire including items on the subject's education, occupation, lifestyle, medical history, exposure to radiation for diagnosis or therapy, head trauma and blood group and the medical history of family members. The series of cerebral tumors was first considered as an indistinct set: none of the risk factors examined showed a statistically significant association. A positive association was found with blood group A (OR = 6) when low-grade astrocytomas (n = 41) were considered separately. As for the malignant astrocytomas (n = 132), there was a suggestive but not statistically significant association with the presence of CNS tumors among first- and second-degree relatives (OR = 7.0). On the whole, this study yielded no clear and meaningful association for the various risk factors analyzed.

[Changes in lymphocyte subsets after radiotherapy]. / Variazione dei subset linfocitari dopo radioterapia.

Radiation therapy seems to induce depletion of lymphocytes, which are very important cells for immunity response. The lymphocyte phenotype was studied in 41 non-pretreated patients with normal immunological parameters who received postoperative radiation therapy for breast, mediastinal or pelvic cancer with at least 50 Gy/25 fractions. The functional immunological assessment was analyzed by Multiskin test (Merieux) too. The lymphocyte phenotype was determined on whole-blood lysate employing an Ortho double-fluorescence cytofluorimeter. All patients, after radiation treatment, exhibited decrement in absolute and percent lymphocyte subpopulations; the Multiskin test demonstrated simultaneous change in skin-test response. The results are highly significant (p 2-tailed area less than 0.0001) for absolute cells count and skin-delayed response test, but percent variations are not significant when verified by t-test.

[Quantitative and functional assay of AT III in neoplastic pathology]. / Aspetto quantitativo e funzionale dell'AT III nella patologia neoplastica.

Our study compares the results obtained through the assay of AT III following two methods, the functional and the immunological one, carried out on plasma of 222 patients affected with solid malignant neoplasia. The purpose of our research was to examine the behaviour of AT III, a protein having a peculiar function in the complex coagulation mechanism. At least among healthy people, this function was and is correlated to its blood concentration. All examined subjects suffered from solid malignant neoplasia either of the respiratory tract or gastro-intestinal tract or sexual organs or urinary tract. From our research patients with liver neoplasia diagnosis have been excluded since the cancer seat might have significant implications on the genesis of coagulation factors and therefore on AT III itself. The age of patients as well as controls - formed by 100 clinically healthy donors-ranged between 30 and 70. At the very beginning patients were divided into groups according to the basic pathology (cancer seat) but then, since no great differences were noted between the groups, all patients were examined irrespective of the type and seat of neoplasia. The followed methods are: a functional method which tends to measure the total AT III capacity assay acting in enzyme excess. S 2238 (Kabi) being the chromogene substratum used; an immunological method which tends to measure the molecule blood concentration taking advantage of its antigenic properties. While on one hand the obtained results have shown a good correlation between the two methods in controls i.c. healthy subjects, on the other hand a significant difference has been noted between the AT III activity and its blood concentration in non healthy subjects as if to prove the interference of neoplasia in the blood coagulation process. In the patients affected with neoplasia the functional as well as the quantitative aspect - which can be considered synonyms in healthy subjects being the one indicative of the other - show that the delicate balance on which the coagulation mechanism is based has been upset.

[Blood coagulation in 222 cases of solid malignant neoplasms]. / Assetto emocoagulativo in 222 casi di neoplasia maligna solida.

We have examined 222 patients with nonterminal solid malignant tumors in order to study the possible variations on the hemocoagulation system due to neoplasia. Only patients whose hemocoagulation system could be proven normal by test made before the malignant neoplasia appeared were taken into consideration. Our study was based on the following tests: platelet count, platelet adhesiveness and aggregation, prothrombin time, thrombin time, thrombotest, antithrombin 3, thrombin-coagulase, activity partial thromboplastin time, fibrinogen, reptilase time. Our study showed there was no relevant difference between the results tests of healthy people taken as points or reference and those of our patients. We did find however a slight but diffuse alteration of all, or almost all, the components of the hemostatic and coagulation system. These variations however were not significant enough to enable us to prove any connection between the neoplasia and the whole hemocoagulation system.

[Changes in plasma ionized calcium levels during fluorescein angiography]. / Variation du calcium ionisé plasmatique au cours de l'angiographie en fluorescence.

The authors have demonstrated a significant decrease of the plasmatic ionized calcium level in 84 patients during retinal fluorangiography likely due to a chemical bond between calcium and fluorescein. The side effects noticed during the procedure were similar to that quoted in the literature; their frequency, however, was not correlated with the decrease of the plasmatic ionized calcium level, even though the magnitude of the decrease was twice as great in the patients who experienced some trouble as in those who did not. This lack of correlation may be related to the too small patients sample. A greater frequency of side effects has been noticed in patients treated by calcium inhibitors.