Erminio Filimberti

Potential role of interleukin-1 as the trigger for diffuse intravascular coagulation in acute nonlymphoblastic leukemia

Abnormalities of coagulation are common in patients with acute nonlymphoblastic leukemia, although the mechanisms involved are unclear, except in a few cases. To investigate the pathogenesis of this coagulopathy, suspensions of purified leukemic cells were prepared and tested for procoagulant activity. Neither the leukemic cells nor their supernatants directly accelerated the clotting of plasma. Since the leukemic cells did not possess direct procoagulant activity, their ability or inability to elaborate a mediator of cellular coagulant properties, interleukin-1, was studied. Leukemic cells from patients with coagulopathy elaborated interleukin-1, and addition of phytohemagglutinin increased interleukin-1 release. In contrast, no interleukin-1 was released, before or after stimulation with phytohemagglutinin, from leukemic cells from patients without coagulopathy. Leukemic cells from another group of patients with abnormalities of coagulation released interleukin-1 only after phytohemagglutinin treatment. In terms of the coagulation mechanism, interleukin-1 containing supernatants from leukemic cell cultures induced the procoagulant receptor tissue factor, a co-factor in the initiation of coagulation, on the endothelial cell surface. There was coordinate suppression of the anticoagulant endothelial cell receptor thrombomodulin, a co-factor for the antithrombotic protein C pathway. Antibody to interleukin-1 prevented these changes in cellular coagulant properties. Taken together, these changes result in a shift in the balance of endothelial cell coagulant properties to an activated state in which mechanisms promoting procoagulant reactions on the vessel surface predominate. Synthesis and release of the mediator interleukin-1 by leukemic cells thus defines a new mechanism through which malignant cells can potentially activate the coagulation mechanism.

The Association between Varicocele, Premature Ejaculation and Prostatitis Symptoms: Possible Mechanisms

INTRODUCTION No study has ever systematically evaluated the impact of varicocele on sexual function. AIM Two cross-sectional studies were performed in patients attending an andrology unit either for male sexual dysfunction (study 1) or couple infertility (study 2). In study 1, we evaluated the impact of varicocele on sexual function. In study 2, we retrospectively evaluated a possible association between varicocele and prostatitis signs and symptoms. METHODS Study 1 refers to a consecutive series of 2,448 (mean age 52.0 +/- 12.9 years) subjects. Study 2 consists of a consecutive series of 139 male subjects (mean age 37.3 +/- 6.3). MAIN OUTCOME MEASURES In study 1, varicocele was clinically classified into three grades according to Dubin criteria. Different hormonal parameters were also evaluated. All the patients of study 2 underwent simultaneous scrotal and transrectal color-Doppler ultrasonography (CDU) along with seminal characteristics and interleukin-8, a surrogate marker of prostatitis. RESULTS After adjusting for age, subjects with severe varicocele (N = 284, 11.6%; Dubin grade 2 and 3) showed a reduction of testicular volume (P < 0.01), higher luteinizing hormone (LH) (P < 0.05), follicle stimulating hormone (FSH) (P < 0.0001) and prolactin (P < 0.05) levels, and also an enlarged or tender prostate at digito-rectal examination (P < 0.05). Premature ejaculation was the only sexual symptoms significantly associated with varicocele (29.2% vs. 24.9% in subjects with or without varicocele, respectively; P < 0.05). In study 2, subjects with severe echographic-defined varicocele (basal venous reflux increasing or not after Valsalvas maneuver; N = 28, 20.1%) showed CDU features of prostatitis and higher seminal inteleukin-8 levels. The presence of any degree of varicocele (N = 40, 28.8%) was also associated with prostatitis symptoms, as measured by the National Institutes of Health Chronic Prostatitis Symptom Index scoring (P < 0.05), and in particular with the pain domain (P < 0.05). CONCLUSIONS In conclusion, signs and symptoms of prostatitis are more common in varicocele patients, who more often complain of premature ejaculation.

Ultrasonographic and clinical correlates of seminal plasma interleukin-8 levels in patients attending an andrology clinic for infertility

This study was aimed at evaluating the association between seminal plasma interleukin-8 (sIL-8) and colour-Doppler ultrasound (CDU) characteristics of the male genital tract in a series of patients fulfilling the criteria of male accessory gland infections (MAGI). Of 250 subjects seeking medical care for couple infertility, 79 (mean age: 36.4 ± 7.5 years) met the criteria of MAGI and scored higher than the rest of the sample on the National Institutes of Health-Chronic Prostatitis Symptom Index score. All patients underwent simultaneous hormone evaluation and seminal analysis (including sIL-8), along with scrotal and transrectal CDU before and after ejaculation. After adjusting for age, sIL-8 in patients with MAGI was significantly related to several abnormal semen and CDU parameters. In particular, leucocytospermia was closely associated with sIL-8. Ejaculate volume, unlike other semen or hormonal parameters, was negatively associated with sIL-8. When scrotal CDU was performed, sIL-8 was positively related to CDU inhomogeneous, hypo-echoic, hyper-echoic epididymis and to epididymal calcifications. In addition, a positive correlation among sIL-8, hyperaemic epididymis and an increased size of epididymal tail was found. When transrectal CDU was performed, an association among sIL-8 and hyper-echoic seminal vesicles, dilated ejaculatory ducts and duct calcifications was also observed. Finally, sIL-8 was positively related to prostate CDU abnormalities such as calcifications, inhomogeneous/hypo-echoic texture, hyperaemia and high arterial blood flow. No association was found with testis parameters. In conclusion, sIL-8 levels in patients with MAGI are associated with several parameters and CDU abnormalities of epididymis, seminal vesicles, ejaculatory ducts and prostate, but not of the testis. Furthermore, sIL-8 positively correlates with CDU signs of ejaculatory duct inflammatory subobstruction.

Elevated body mass index correlates with higher seminal plasma interleukin 8 levels and ultrasonographic abnormalities of the prostate in men attending an andrology clinic for infertility

Background: Obesity is associated with a systemic, low-grade inflammatory state. Although the relationship between obesity and semen parameters or prostate diseases has been previously investigated, the association between body mass index (BMI), prostate inflammatory diseases and color-Doppler ultrasound (CDU) of the male genital tract (MGT) has been poorly studied. Aim: To evaluate the association between BMI and CDU features of the MGT, signs and symptoms of prostate inflammation, semen parameters. Materials/Subjects and methods: We studied 222 men seeking medical care for couple infertility. According to the World Health Organization classification, subjects were divided into 3 groups: normal weight (no.=131, BMI=18.5–24.9 kg/m2), overweight (no.=71, BMI=25.0–29.9 kg/m2), obese (no.=20, BMI≥30.0 kg/m2). All patients underwent simultaneous testosterone evaluation and seminal analysis, including interleukin 8 (sIL-8), along with scrotal and transrectal CDU, before and after ejaculation. Prostatitis symptoms were evaluated by National Institutes of Health-Chronic Prostatitis Symptom Index questionnaire. Results: After adjusting for age and testosterone levels, higher BMI was significantly related to higher prostate volume and several CDU features of the prostate, including macro-calcifications, inhomogeneity, higher arterial peak systolic velocity (the latter adjusted also for blood pressure), but not with abnormalities of testis, epididymis, seminal vesicles. Furthermore, higher BMI and BMI class were significantly related to higher sIL-8, a reliable surrogate marker of prostate inflammatory diseases, even after adjustment for age. Conversely, no associations among BMI, clinical symptoms of prostatitis or semen parameters were observed. Conclusions: Subjects with higher BMI might develop CDU and biochemical signs suggestive of prostate inflammation, although not clinically overt.

Multi-variate analysis of factors governing the pharmacokinetics of exogenous factor VIII in haemophiliacs

SummaryThe pharmacokinetics of Factor VIII was evaluated by mathematical modeling in a large-scale study in 62 haemophilia-A subjects, in whom 137 plasma Factor VIII-time curves were measured during single dose (n=87) and repeated-dose (n=47) treatments for prophylaxis or minor bleeding episodes. The pharmacokinetic parameters [mean (SD)] estimated from single-dose curves were: clearance 3.85 ml·h−1·kg−1, volume of distribution 58.2 ml·kg−1, mean residence time 15.9 h. Parameters calculated from repeated-dose curves were: clearance 3.93 ml·h−1·kg−1, volume of distribution 61.8 ml·kg−1, and half-life 12.2 h. In patients with mild haemophilia, pharmaco-statistical analysis revealed that the endogenous synthesis of Factor VIII was constant and was not influenced by the administration of exogenous Factor VIII. The coefficient of variation for intra-individual variability of Factor VIII kinetics (estimated according to the Standard Two-Stage method) was 20.7% in single-dose curves and 23.2% in repeated-dose curves.

Sperm DNA fragmentation induced by cryopreservation: new insights and effect of a natural extract from Opuntia ficus-indica

OBJECTIVE To analyze the effect of cryopreservation on sperm DNA fragmentation (SDF) in two cytometric sperm populations, PI(brighter) and PI(dimmer), and to test the effects of Opuntia ficus-indica (OFI) extracts, which contain antioxidants and flavanoids, and of resveratrol on cryopreservation of human semen. DESIGN In vitro prospective study. SETTING Institutional study. PATIENT(S) Twenty-one normozoospermic men undergoing semen analysis for couple infertility. INTERVENTION(S) Cryopreservation using the routine method in the presence of OFI extracts or resveratrol. MAIN OUTCOME MEASURE(S) Measurement of SDF by TUNEL/PI flow cytometric method to evaluate sperm motility (by automated motion analysis, CASA system) and viability (by eosin/nigrosin staining) in the two populations of sperm PI(br) and PI(dim). RESULT(S) Cryopreservation induced an increase of SDF only in the PI(br) sperm population. The increase was negatively dependent on the basal values of SDF in the same population. Addition of OFI extracts and resveratrol to the cryopreservation medium slightly but statistically significantly reduced SDF in the PI(br) population without affecting the deleterious effect of cryopreservation on sperm motion parameters or viability. CONCLUSION(S) The increase of SDF in the PI(br) population, which is unrelated to semen quality, suggests that caution must be taken in using cryopreserved semen, as morphologically normal and motile sperm may be damaged. The addition of substances with multifunctional properties such as OFI extracts to cryopreservation medium is only slightly effective in preventing the dramatic effects on SDF.

Acrosome reaction is impaired in spermatozoa of obese men: a preliminary study

OBJECTIVE To compare spontaneous (Sp-AR) and P-induced acrosome reaction (AR) in spermatozoa of obese and lean subjects. SETTING Bariatric unit at a university hospital. DESIGN Prospective, observational study. PATIENT(S) Twenty-three obese (mean±SD body mass index [BMI], 44.3±5.9 kg/m2) and 25 age-matched lean (BMI, 24.2±3.0 kg/m2) subjects. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Spontaneous and P-induced AR in spermatozoa of obese and lean subjects. RESULT(S) A statistically significant difference was found between obese and lean cohorts in total T and calculated free T, E2, glycated hemoglobin, and high-density lipoproteins, whereas among the routine semen parameters analyzed, only immotile sperm percentage and ejaculate volume differed significantly. Spermatozoa of obese (n=13) vs. lean men (n=19) showed a higher Sp-AR (17.9%±7.2% vs. 8.3%±4.2%), which resulted in a reduced ability to respond to P evaluated as the AR-after-P-challenge parameter (3.5%±3.2% vs. 17.6%±9.2%). Multivariate analysis adjusted for age revealed a significant correlation between BMI, waist, E2, and glycated hemoglobin with both Sp-AR (age-adjusted r=0.654, r=0.711, r=0.369, and r=0.644, respectively) and AR-after-P-challenge (age-adjusted r=-0.570, r=-0.635, r=-0.507, and r=-0.563, respectively). A significant difference in sperm cholesterol content was reported between obese and lean men (29.8±19.5 vs. 19.1±14.6 ng/μg of proteins). CONCLUSION(S) Sperm AR is impaired in obese men, showing reduced response to P and elevated Sp-AR, associated with altered circulating levels of E2 and sperm cholesterol content.

High variability in results of semen analysis in andrology laboratories in Tuscany (Italy): the experience of an external quality control (EQC) programme

We report the results of the first three trials of an external quality control (EQC) programme performed in 71 laboratories executing semen analysis in Tuscany Region (Italy). At the end of the second trial, participants were invited to attend a teaching course illustrating and inviting to adhere to procedures recommended by WHO (V edition). Results of the first three trials of the EQC documented a huge variability in the procedures and the results. The highest variability was found for morphology (CV above 80% for all the trials), followed by count (CV of about 60% for all the trials) and motility (CV below 30% for all the trials). When results of sperm count and morphology were divided according to the used method, mean CV values did not show significant differences. CV for morphology dropped significantly at the third trial for most methods, indicating the usefulness of the teaching course for morphology assessment. Conversely, no differences were observed after the course for motility and for most methods to evaluate count, although CV values were lower at the second and third trial for the laboratories using the Burker cytometer. When results were divided according to tertiles of activity, the lowest mean bias values (difference between each laboratory result and the median value of the results) for count and morphology were observed for laboratories in the third tertile (performing over 200 semen analysis/year). Of interest, mean bias values for concentration dropped significantly at the third trial for low activity laboratories. In conclusion, lack of agreement of results of semen analysis in Tuscany is mainly because of the activity and the experience of the laboratory. Our study points out the importance of participating in EQC programmes and periodical teaching courses as well as the use of WHO recommended standardized procedures to increase precision and to allow the use of WHO reference values.

Single-dose pharmacokinetics of factor IX evaluated by model-independent methods

We studied the pharmacokinetic data of 13 subjects with hemophilia B treated with a single‐dose of a Factor IX concentrate (Bebulin TIM2, N = 9; Preconativ, N = 4). The decay curves of Factor IX were evaluated by model‐independent methods and the following pharmacokinetic parameters (mean ± SD) were estimated: clearance (ml/h/kg) = 4.99 ± 2.01; mean residence time (h) = 22.9 ± 10.6; volume of distribution (ml/kg) = 99.9 ± 35.5. The in vivo recovery (59.8% ± 16.9%) was found to be inversely correlated with the volume of distribution. No significant difference in the pharmacokinetic parameters was found between patients treated with Preconativ and those treated with Bebulin. A model‐dependent compartmental evaluation of the 13 decay curves showed that the two‐compartment model was better than the one‐compartment model in 7 cases (53.8%), but the improvement of fit resulting from the two‐compartment model was statistically significant in only 2 cases (15.4%).

Semen cryopreservation for men banking for oligospermia, cancers, and other pathologies: prediction of post-thaw outcome using basal semen quality

OBJECTIVE To evaluate post-thawing sperm parameters in a large series of men cryopreserving for different cancers and oligospermia. DESIGN Retrospective observational study. SETTING Semen cryopreservation laboratory. PATIENT(S) Six hundred twenty-three patients undergoing semen cryopreservation for cancer or oligospermia who discontinued banking. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Postcryopreservation sperm motility and viability. RESULT(S) In oligospermic men, recovery of motile sperm after cryopreservation was possible in only a few out of the 219 samples cryopreserved for this problem. Similarly, independent of the reason for which cryopreservation was required, if one basal semen parameter fell below the 5th percentile of the World Health Organization reference values, recovery of motile and viable spermatozoa after thawing was low. Among samples cryopreserved for cancer, those with testicular cancer showed the lowest basal semen quality and recovery after thawing. In cases of hematological cancers or other types of cancers, motility recovery was similar to that of non-cancer-related samples. Receiver operating characteristic analyses demonstrate that basal progressive and total motility predict the recovery rate of motile sperm after thawing with high accuracy, sensibility and specificity. CONCLUSION(S) Our study demonstrates the ability of prefreeze semen parameters to predict cryosurvival in terms of sensitivity and precision. Using this information, the clinician could perform appropriate counseling about the future possibilities of fertility for the patient.