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Background & Objectives: Chronic social stress is an important factor responsible for the worsening of depressive disorders in humans. In this study we present the relational Neuro-Psycho-Physical Optimization (NPPO) with Radio Electric Asymmetric Conveyer (REAC-CRM) as the treatment to tackle the unconscious dysfunction adjustments carried out by the central nervous system as a response to environmental stresses., Methods: Psychological stress was measured in a group of 888 patients using the Psychological Stress Measure (PSM) test, a self-administered questionnaire. Data were collected immediately before and after the 4-wk therapy cycle. The detection of anxiety and depression clusters by PSM test has been based on the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR, APA, 2000) criteria. Six hundred and eighty eight patients (212 males, 476 females, average PSM test total scores 107.9 +/- 23.13) were treated with REAC-CRM therapy; 200 (64 males, 136 females, average PSM test total scores 107.86 +/- 25.80) were treated with "placebo REAC-CRM therapy"and used as control., Results: This study showed a significant reduction in scores measuring subjective perceptions of stress in the patients treated with a cycle of REAC-CRM therapy. At the end-point the number of patients reporting symptoms of stress-related anxiety and depression on the PSM test was significantly reduced (P<0.001); in the placebo group no significant difference was highlighted., Interpretation & Conclusions: NPPO therapy with a cycle of REAC-CRM was shown to reduce subjective perceptions of stress measured by the PSM test and in particular, symptoms of stress-related anxiety and depression.

This retrospective study was aimed at evaluating the effects of cigarette consumption on semen parameters in a group of men with idiopathic infertility. The semen quality of 2 groups of men with idiopathic infertility, smokers (n = 118) and nonsmokers (n = 153), were compared. Conventional semen analysis was performed and sperm morphology was assessed by transmission electron microscopy (TEM). TEM data were elaborated by means of a mathematical formula based on a Bayesian technique able to furnish a fertility index (FI), and the percentages of sperm apoptosis, necrosis, and immaturity. Values of normality recommended by World Health Organization guidelines were used as a control for conventional semen analysis, and values from sperm of 25 men of proven fertility were used for TEM indices. Infertile smoker and nonsmoker patients showed similar sperm parameters, although sperm motility and TEM analysis values in both groups were significantly impaired compared with controls. Smoker patients were then classified as mild (>or=1 and 10 and <20 cigarettes/day), or heavy smokers (>or=20 cigarettes/d). Sperm concentration and FI were significantly (P < .05) different among the 3 considered smoker classes. Comparing the pairs of smoker classes, sperm concentration and FI in heavy smokers were significantly lower (P < .05) than that observed in mild smoker and nonsmoker groups. Although semen quality in males with idiopathic infertility seems not to be dramatically affected by cigarette consumption, heavy smokers show significantly lower sperm concentration and FI: another strong reason to stop smoking.

Estimates of semen parameters are important for defining normal ranges, which are currently established by 1999 World Health Organization guidelines. However, it is well known that semen evaluation is questionable because it is necessary for only 1 sperm to be able to reach and fertilize the oocyte. Spermiogram parameters and sperm morphology, evaluated by transmission electron microscopy (TEM), were performed on semen samples from 25 men of proven fertility. Despite a generally normal sperm concentration, progressive motility was reduced in 9 cases. Sperm characteristics were evaluated with an established technique, and the mean of the percentages of sperm pathologies were confirmed by comparing to previous reports. A comparison of apoptosis and necrosis in the samples, as detected by TEM and an annexin V/propidium iodide assay, was also performed. Fluorescence in situ hybridization was carried out on the same samples using probes for chromosomes 18, X, and Y. The mean value of the frequency of total aneuploidy in the analyzed group was 0.627% (25th percentile = 0.563%; median = 0.625%; 75th percentile = 0.690%). This study of the incidence of disomy and diploidy in spermatozoa from fertile, apparently normal individuals is important for making comparisons with infertile cohorts to determine the real increase of aneuploidy in those cohorts.

Aim: To perform screening, related to A-kinase anchoring proteins 4 (AKAP4) and tubulin proteins, in spermatozoa with absent or severely reduced motility in order to detect the status of the fibrous sheath and the axonemal structure., Methods: An immunocytochemical study of tubulin, used as a positive control, and AKAP4 was carried out to detect the presence and the distribution of these proteins in different sperm samples. The morphological characteristics of sperm were studied by transmission electron microscope (TEM) and the results were elaborated using a formula reported in previous studies. PCR was carried out on DNA extracted from peripheral blood lymphocytes to analyse partial sequences of the Akap4 and Akap3 genes., Results: Immunolabelling of tubulin and AKAP4 showed different patterns, which led us to divide the patients into groups. In group I, the absence of AKAP4 and tubulin was revealed, although these patients did not show alterations in the Akap4/Akap3 binding site. TEM evaluation highlighted that a high presence of necrosis was associated with total sperm immotility. In group II, a regular AKAP4 and tubulin signal was present, although motility was reduced and TEM analysis revealed the presence of immaturity. In group III, in which a weak AKAP4 label associated with normal tubulin staining and reduced motility was observed, a severe disorganization of the fibrous sheath was highlighted by TEM., Conclusion: While the role of AKAP4 in sperm motility is unclear, absent or weak AKAP4-labelling seems to be associated with absent or weak sperm motility.

Finasteride is a specific inhibitor of the 5alpha reductase enzyme originally approved for the treatment of benign prostatic hypertrophy and also for the treatment of androgenetic alopecia (AGA) in men at a dose of 1 mg/day. We report on three cases of young men recruited at our Centre for Male Infertility who had used finasteride for five years. Semen quality was investigated by light microscopy to evaluate sperm concentration and motility. Sperm morphology was performed by transmission electron microscope (TEM) and the data were analyzed. The presence of Y microdeletions was investigated by PCR. Meiotic segregation was explored by fluorescence in situ hybridization (FISH). Patient 1 was azoospermic, patients 2 and 3 showed a normal sperm concentration and severely reduced progressive motility. TEM analysis revealed altered sperm morphology consistent with necrosis and FISH data revealed elevated diploidy and sex chromosome disomy frequencies. This examination was repeated 1 year after the men had suspended the use of finasteride, without receiving any other treatment. A recovery of spermatogenetic process was observed. Motility and morphology improved whereas the meiotic pattern did not change presenting elevated diploidy and sex chromosome disomy frequency.

Cryptorchidism is a pathological condition defined as the failure of the testis to descend into the scrotum, the location of the cryptorchid testis can be in the inguinal canal or in the prescrotal and abdominal area, sometimes resulting in atrophic seminiferous tubules. The aim of this study was to analyze semen quality of men who underwent orchidopexy for unilateral or bilateral cryptorchidism during childhood. Semen quality was investigated by light microscopy to evaluate sperm concentration and motility. Sperm morphology was performed by transmission electron microscope (TEM), and the data were mathematically elaborated. The presence of Y microdeletions was investigated by polymerase chain reaction. The effect of cryptorchidism on meiosis was explored by fluorescence in situ hybridization (FISH). The incidence of azoospermia was higher in the group with bilateral compared with unilateral cryptorchidism, and semen parameters were better in the unilateral group. Sperm pathologies detected by TEM indicated a severe deterioration of sperm quality in both groups. Necrosis and apoptosis appeared to be the most frequent pathologies, and their values reached statistical significance compared with those from fertile controls. The presence of chromosome Y microdeletions in patients with cryptorchidism and severe spermatogenetic defects is controversial. No microdeletions were found in this study. FISH values indicated that the mean percentage of gonosome disomies and diploidies were generally out of normal range, indicating a severe disturbance of meiotic segregation. The effects induced by cryptorchidism resolved in childhood seem to include a spermatogenetic impairment, leading to recommendation of detailed ultrastructural and chromosomal sperm analyses before undertaking assisted reproductive techniques.

The idea that varicocele plays a detrimental role in fertility is supported by the presence of a higher frequency of affected men among the infertile population than among men with normal semen parameters. In this research we examined ejaculates from a large group of selected men affected by varicocele by light and electron microscopy. The effect of varicocele on chromosome meiotic segregation was investigated by fluorescence in situ hybridization (FISH). The potential benefits of varicocelectomy on sperm quality were evaluated by analyzing sperm characteristics before and after surgical correction of varicocele. Transmission electron microscopy (TEM) analysis, elaborated previously, showed that the incidence of immaturity, apoptosis, and necrosis was higher in the varicocele group than in controls. FISH analysis performed on sperm nuclei from selected patients with varicocele showed that the mean frequencies of disomies and diploidies were generally out of the normal range, indicating a severe disturbance in meiotic segregation. Sperm characteristics evaluated before and after varicocele repair showed a general improvement. As a consequence, the varicocele seem to affect sperm morphology and function concomitantly with meiotic segregation derangement. In consideration of these data, we suggest that TEM and FISH analyses should be performed for all varicocele patients.

Introduction: Current practices for renal replacement therapy in intensive care units (ICUs) remain poorly defined. The DOse REsponse Multicentre International collaborative initiative (DO-RE-MI) will address the issue of how the different modes of renal replacement therapy are currently chosen and performed. Here, we describe the study protocol, which was approved by the Scientific and Steering Committees., Methods: DO-RE-MI is an observational, multicentre study conducted in ICUs. The primary end-point will be the delivered dose of dialysis, which will be compared with ICU mortality, 28-day mortality, hospital mortality, ICU length of stay and number of days of mechanical ventilation. The secondary end-point will be the haemodynamic response to renal replacement therapy, expressed as percentage reduction in noradrenaline (norepinephrine) requirement. Based on the the sample analysis calculation, at least 162 patients must be recruited. Anonymized patient data will be entered online in electronic case report forms and uploaded to an internet website. Each participating centre will have 2 months to become acquainted with the electronic case report forms. After this period official recruitment will begin. Patient data belong to the respective centre, which may use the database for its own needs. However, all centres have agreed to participate in a joint effort to achieve the sample size needed for statistical analysis., Conclusion: The study will hopefully help to collect useful information on the current practice of renal replacement therapy in ICUs. It will also provide a centre-based collection of data that will be useful for monitoring all aspects of extracorporeal support, such as incidence, frequency, and duration.

Objective: To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS)., Design: Retrospective study., Setting: Regional Referral Center for Male Infertility, Siena, Italy., Patient(s): Twelve infertile patients with DFS sperm defects., Intervention(s): Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis., Main Outcome Measure(s): The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction., Result(s): The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls., Conclusion(s): These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.

Objective: To characterize and describe the ontogenesis of a rare flagellar defect affecting the whole sperm population of a sterile man., Design: Case report., Setting: Regional referral center for male infertility in Siena, Italy., Patient(s): A 28-year-old man with severe asthenozoospermia., Intervention(s): Physical and hormonal assays, semen analysis, and testicular biopsy., Main Outcome Measure(s): Semen samples and testicular biopsies were analyzed by light and transmission electron microscopy; immunocytochemical study with anti-beta-tubulin and anti-AKAP 82 antibodies was performed to detect the presence and distribution of proteins., Result(s): Ultrastructural analysis of ejaculated spermatozoa and testicular biopsy revealed absence of the fibrous sheath in the principal-piece region of the tail. Fibrous sheath-like structures were observed in cytoplasmic residues and residual bodies released by spermatids in the seminiferous epithelium. Other anomalies observed were supplementary axonemes and mitochondrial helix elongation. These features were confirmed by immunocytochemical staining., Conclusion(s): This rare sperm tail defect, characterized by absence of the fibrous sheath, presence of supplementary axonemes, and an abnormally elongated midpiece, originates in the seminiferous tubules during spermiogenesis, as detected in testicular biopsy sections. These defects occur in the whole sperm population, and therefore a genetic origin could be suggested.

Unlabelled: The preparation, characterization, and first biologic evaluation in rats of a novel class of monocationic (99m)Tc heart imaging agents are reported. The complexes are represented by the general formula [(99m)Tc(N)(PNP)(L)](+), where L is the monoanionic form of a dithiocarbamate ligand of the type [R(1)(R(2))-N-C(=S)S](-), PNP is a diphosphine ligand of the type [(R(3))(2)P-(CH(2))(2)](2)-N(R(4)), and R(1)-R(4) are organic functional groups., Methods: The new complexes were prepared by use of both liquid and freeze-dried formulations through a 2-step procedure. The first step involved the formation of the [TcN](2+) group through the reaction of (99m)TcO(4)(-) with succinic dihydrazide as a donor of nitride nitrogen atoms (N(3-)) in the presence of Sn(2+) ions. This step was followed by the simultaneous addition to the reaction solution of the ligand PNP and of the sodium salt of the dithiocarbamate ligand (NaL) to afford the final products, [(99m)Tc(N)(PNP)(L)](+). The chemical identities of the resulting (99m)Tc complexes were determined by comparing their chromatographic properties with those of the corresponding (99g)Tc analogs prepared by use of the long-lived isotope (99g)Tc and characterized by spectroscopic and crystallographic techniques. Ex vivo biodistribution studies were conducted in rats. In vivo tomographic images of the rat heart were obtained by use of a small-animal SPECT scanner., Results: The [(99m)Tc(N)(PNP)(L)](+) complexes are monocationic and possess a distorted square-pyramidal geometry in which the TcN multiple bond occupies an apical position and the diphosphine and dithiocarbamate ligands span the residual 4 coordination positions on the basal plane through the 2 phosphorus atoms and the 2 sulfur atoms, respectively. Imaging and biodistribution studies demonstrated that these radiopharmaceuticals localize selectively in the myocardium of rats and are retained in this region for a prolonged time. The kinetics of heart uptake and clearance were found to be influenced by variations in the lateral R(1)-R(4) groups. Blood and lung washouts were extremely fast. Elimination occurred mostly through the kidneys and the liver. Surprisingly, at 1 h after injection, liver activity was almost negligible. Analysis of heart-to-liver and heart-to-lung uptake ratios showed that these values increased exponentially over time and became much higher than those determined for (99m)Tc-sestamibi and (99m)Tc-tetrofosmin. These findings were confirmed by analysis of high-quality SPECT images collected in rats for the new complexes and compared with images obtained with (99m)Tc-sestamibi and (99m)Tc-tetrofosmin., Conclusion: The high myocardial uptake and the very high heart-to-lung and heart-to-liver uptake ratios indicate that the [(99m)Tc(N)(PNP)(L)](+) complexes exhibit very favorable distribution properties and could be used to obtain SPECT cardiac images with improved quality.