Lung visualisation on PET/MRI: implementing a protocol with a short echo-time and low flip-angle volumetric interpolated breath-hold examination sequence.

AIM: To assess the diagnostic performance in detecting lung lesions of a short echo-time (TE) and low flip-angle (FA) volumetric interpolated breath-hold examination (VIBE) sequence included in the integrated positron-emission tomography (PET)/magnetic resonance imaging (MRI) protocol. METHOD AND MATERIALS: Thirty-seven oncological patients who underwent computed tomography (CT) and PET/MRI, including both a dedicated short TE, low FA VIBE (modified VIBE) and a standard VIBE of the lung, were enrolled. Modified VIBE images were reviewed retrospectively and independently by three raters, to detect pulmonary nodules, parenchymal consolidation, and bands. Three other groups examined standard VIBE, PET, and CT images. MRI and PET findings were compared to CT using Krippendorff's alpha using patient-based and a lesion-based analysis. Krippendorff's alpha was calculated to assess the interobserver agreement among the three raters of the modified VIBE. RESULTS: In the patient-based analysis (positivity ≥1 lesion), the comparison of modified VIBE with CT showed an alpha of 0.54 for nodules <6 mm (versus 0.41 for standard VIBE and 0.09 for PET) and an alpha of 0.88 for nodules ≥6 mm (versus 0.74 for standard VIBE and 0.42 for PET). On a lesion-based analysis (presence/absence of each lesion), modified VIBE compared to CT showed an alpha of0.58 for nodules <6 mm (versus 0.44 for standard VIBE and 0.09 for PET) and an alpha of 0.90 for nodules ≥6 mm (versus 0.79 for standard VIBE and 0.50 for PET). The alpha value for the interobserver agreement was 0.90 for nodules <6 mm, 0.91 for nodules ≥6 mm, 1.00 for consolidations, and 0.95 for bands in the patient-based analysis and 0.89, 0.93, 1.00, and 0.95 in the lesion-based analysis. CONCLUSIONS: Modified VIBE proved to be reproducible, showed better accuracy than standard VIBE and PET, and very good concordance with CT in assessing lung nodules ≥6 mm, whereas the agreement was less satisfactory for smaller nodules.

Effect of microparticulated whey proteins on milk coagulation properties.

The enhancement of milk coagulation properties (MCP) and the reuse of whey produced by the dairy industry are of great interest to improve the efficiency of the cheese-making process. Native whey proteins (WP) can be aggregated and denatured to obtain colloidal microparticulated WP (MWP). The objective of this study was to assess the effect of MWP on MCP; namely, rennet coagulation time (RCT), curd-firming time, and curd firmness 30 min after rennet addition. Six concentrations of MWP (vol/vol; 1.5, 3.0, 4.5, 6.0, 7.5, and 9.0%) were added to 3 bulk milk samples (collected and analyzed during 3 d), and a sample without MWP was used as control. Within each day of analysis, 6 replicates of MCP for each treatment were obtained, changing the position of the treatment in the rack. For control samples, 2 replicates per day were performed. In addition to MCP, WP fractions were measured on each treatment during the 3 d of analysis. Milk coagulation properties were measured on 144 samples by using a Formagraph (Foss Electric, Hillerød, Denmark). Increasing the amount of MWP added to milk led to a longer RCT. In particular, significant differences were found between RCT of the control samples (13.5 min) and RCT of samples with 3.0% (14.6 min) or more MWP. A similar trend was observed for curd-firming time, which was shortest in the control samples and longest in samples with 9.0% MWP (21.4 min). No significant differences were detected for curd firmness at 30 min across concentrations of MWP. Adjustments in cheese processing should be made when recycling MWP, in particular during the coagulation process, by prolonging the time of rennet activity before cutting the curd.

Vascular risk factors for primary open angle glaucoma: the Egna-Neumarkt Study.

OBJECTIVE: To assess the impact of vascular risk factors on the prevalence of primary open angle glaucoma. DESIGN: Population-based cross-sectional study. PARTICIPANTS: Four thousand two hundred ninety-seven patients more than 40 years of age underwent a complete ocular examination in the context of the Egna-Neumarkt Glaucoma Study. INTERVENTION: Ocular examinations were performed by trained, quality-controlled ophthalmologists according to a predefined standardized protocol including medical interview, blood pressure reading, applanation tonometry, computerized perimetry, and optic nerve head examination. MAIN OUTCOME MEASURES: Prevalences of ocular hypertension, primary open-angle glaucoma, normal-tension glaucoma, and other types of glaucoma were determined. Correlation coefficients were calculated for the association between systemic blood pressure and age-adjusted intraocular pressure (IOP) and between age and both intraocular and systemic blood pressures. Odds ratios were computed to assess the risk of primary open-angle glaucoma and normal-tension glaucoma in relation to systemic hypertension or antihypertensive medication, blood pressure levels, diastolic perfusion pressure, and a number of other cardiovascular risk factors. RESULTS: A positive correlation was found between systemic blood pressure and IOP, and an association was found between diagnosis of primary open-angle glaucoma and systemic hypertension. Lower diastolic perfusion pressure is associated with a marked, progressive increase in the frequency of hypertensive glaucoma. No relationship was found between systemic diseases of vascular origin and glaucoma. CONCLUSIONS: Our data are in line with those reported in other recent epidemiologic studies and show that reduced diastolic perfusion pressure is an important risk factor for primary open-angle glaucoma.

Epidemiology of angle-closure glaucoma: prevalence, clinical types, and association with peripheral anterior chamber depth in the Egna-Neumarket Glaucoma Study.

OBJECTIVE: To assess the prevalence of primary angle-closure glaucoma (PACG), the frequency of its different clinical presentations, and its association with peripheral anterior chamber depth in a defined population in Northern Italy. DESIGN: Cross-sectional epidemiologic study in a defined population. PARTICIPANTS: All subjects resident in the Egna-Neumarkt area of the South Tyrol Region (Northern Italy) and more than 40 years of age were invited to undergo an ophthalmologic examination. INTERVENTION: After the screening examination, subjects with suspected glaucoma were re-examined at the screening center to confirm the diagnosis. All cases that still proved suspect after the second examination underwent a third phase of investigations and were classified as healthy or as definitely glaucomatous. Each subject was examined according to a standard protocol, including medical history interview, refraction and visual acuity determination, ocular biomicroscopy, evaluation of peripheral anterior chamber depth by means of the Van Herick method, applanation tonometry, optic disc evaluation, and computerized perimetry. Gonioscopy was not performed during initial screening but only in all selected patients in the second and third phases of investigations. The diagnosis of PACG was made on the basis of the concomitant presence of at least two of the following criteria: intraocular pressure > or = 22 mmHg, glaucomatous optic disc abnormalities, glaucomatous visual field defects. In addition, biomicroscopic or gonioscopic evidence of angle closure was also necessary. MAIN OUTCOME MEASURES: Percentage distribution of peripheral anterior chamber depths, prevalence of angle-closure glaucoma, and frequency of the different PACG clinical presentations. RESULTS: Four thousand two hundred ninety-seven subjects were examined (73.9% overall participation rate). The peripheral depth of the anterior chamber according to the Van Herick method was grade 2 in 14.7%, grade 1 in 2.5%, and grade 0 in 0.3% of the population. The overall prevalence of angle-closure glaucoma was 0.6% (26 cases). Five of these were cases of previous acute attacks resolved by therapy, three were cases of chronic angle-closure after acute attacks, three were intermittent angle-closure glaucomas, and 15 were chronic angle-closure cases. CONCLUSIONS: Occludable angles were more frequent than in other white populations previously studied. The prevalence of PACG is not as low as is usually believed; this type of glaucoma accounts for more than a quarter of all glaucomas found in the Egna-Neumarkt population. The most frequent clinical presentation is chronic angle-closure glaucoma.

Prevalence of glaucoma and intraocular pressure distribution in a defined population. The Egna-Neumarkt Study.

OBJECTIVE: The purpose of the study is to assess the prevalence of various types of glaucoma and to determine the intraocular pressure (IOP) distribution in a defined population in an Italian rural community. DESIGN: A cross-sectional epidemiologic study in a defined population was planned. After the screening examination, the subjects with suspected glaucoma were re-examined at the screening center in order to confirm the diagnosis. All cases that still proved suspect after the second examination underwent a third phase of investigations at the Ophthalmologic Division of Bolzano Hospital, and were classified as healthy or definitely glaucomatous. PARTICIPANTS: All subjects residing in the Egna-Neumarkt area of Alto Adige region (Northern Italy) and over 40 years of age were invited to undergo an ophthalmologic examination. INTERVENTION: Each subject was examined according to a standard protocol, including computerized perimetry, applanation tonometry, evaluation of anterior chamber depth and optic disc, and a medical history interview. The diagnosis of glaucoma was based on the presence of at least two of the following criteria: IOP > or = 22 mmHg, glaucomatous optic disc abnormalities, and glaucomatous visual field defects. Ocular hypertension was defined as IOP > or = 22 mmHg without visual field or glaucomatous optic disc abnormalities. MAIN OUTCOME MEASURES: Participation rate, mean IOP, prevalence of glaucoma (primary open-angle glaucoma, primary angle-closure glaucoma, normal-tension glaucoma, secondary glaucoma), and ocular hypertension were determined. RESULTS: Of a total of 5816, 4297 subjects were examined (73.9% overall participation rate). Intraocular pressure showed a Gaussian-like distribution curve skewed to the right. Mean IOP increased with age, and was slightly higher in men (15.14 mmHg) than in women (14.94 mmHg). The overall prevalences of ocular hypertension, primary open-angle glaucoma, primary angle-closure glaucoma, and normal-tension glaucoma were 2.1%, 1.4%, 0.6%, and 0.6%, respectively. Only 28 of 210 patients with glaucoma or ocular hypertension had been diagnosed prior to the screening. CONCLUSIONS: The distribution of IOP and the prevalence of the different types of glaucoma were similar to those found in other white populations.

Glaucoma and quality of the life.

A questionnaire was prepared and given to 332 subjects who passed through our Institute for Glaucoma with the aim of evaluating their quality of life. The diagnosis of glaucoma is associated with a diminished quality of life in a considerable percentage of the interviewed subjects. The patients are troubled most of all by the inconvenience of the treatments and the fear of visual compromission. Mitotic therapies, polytherapies and systemic therapies are the less agreeable of them. Of the therapy in itself, the most disagreeable aspects are the frequency and timetable of the medications. No quality of life differences were mentioned by surgically treated patients nor those on solely pharmacological treatment.

Diagnostic and clinical features in azoospermia.

BACKGROUND AND OBJECTIVE: The recent advances in assisted fertilization and gamete micromanipulation techniques have enabled fertilization in some forms of azoospermia; for example, epididymal sperm aspiration in obstructive azoospermia. Therefore knowledge of the specific degree of testicular damage is of primary importance, since other clinical parameters, such as FSH plasma levels and testicular volume, do not discriminate between the different testiculopathies. In order to further characterize the specific testicular conditions present in azoospermia, we have examined a large group of azoospermic subjects on the basis of testicular cytological analysis obtained by fine needle aspiration. DESIGN AND PATIENTS: One hundred and twenty-two infertile, azoospermic men were studied by physical examination, FSH radioimmunoassay, testicular ultrasound examination and fine needle aspiration of the testes. Thirty-five infertile normozoospermic subjects were studied as controls. RESULTS: The cytological analysis identified five different sub-types in azoospermic subjects: I, Sertoli cell-only syndrome; II, hypospermatogenesis; III, spermatogonial and/or spermatocytic arrest; IV, spermatidic arrest; and V, normal germ line. The testicular volume was reduced in groups I and II, while the FSH plasma levels were increased in groups I, II and III, suggesting a primary role of spermatids in the control of FSH secretion. CONCLUSIONS: In azoospermic subjects, testicular cytological analysis permits the identification of different sub-types and this classification may be very important in determining therapy, particularly the choice between surgical treatment and the hypothetical use of assisted fertilization techniques by retrieval of epididymal or intratesticular spermatozoa or spermatids.

Immunocytochemical localization of epidermal growth factor receptors in human testis from infertile subjects.

OBJECTIVE: To investigate the immunolocalization of the epidermal growth factor receptor (EGFR) in normal and pathological human testis by immunocytochemical technique. DESIGN: Cytologic specimens were obtained by bilateral fine needle aspiration (FNA) of the testis and stained in May Grünwald-Giemsa for the cytologic analysis; immunolocalization of EGFR was analyzed on duplicate slides from each testis using two anti-EGFR monoclonal antibodies and peroxidase-antiperoxidase technique. SETTING: Infertility center of an academic unit. PATIENTS: A total of 42 infertile patients, affected by various testicular diseases. The control group was made up of 10 normal sperm patients with autoimmune infertility and cytologic picture of normal spermatogenesis. INTERVENTIONS: Exogenous FSH was administered 75 IU IM on alternate days for 3 months on 16 of the infertile patients who showed oligospermia and normal FSH plasma levels. Semen analysis and testicular FNA (and after cytologic and immunocytochemical studies) were repeated at 3 months of treatment. MAIN OUTCOME MEASURES: Luteinizing hormone and FSH plasma levels were determined by RIA methods; qualitative and quantitative parameters for the cytologic evaluation are reported in our previous works. RESULTS: The cytologic analysis permitted identification of seven classes of infertile subjects, characterized by different cytologic pictures. Epidermal growth factor receptor immunostaining evidenced weak positivity on Sertoli and germ cells (with the exception of spermatozoa) in the presence of normal germ line and normal FSH plasma levels and strongly intense positivity in the presence of serious hypospermatogenesis, spermatogonial or spermatocytic arrest, and Sertoli cell-only syndrome. These conditions were characterized by higher FSH plasma levels than normal controls. All of the subjects who received exogenous FSH, with moderate hypospermatogenesis or spermatidic arrest, showed on Sertoli and germ cells a weak EGFR immunostaining before the treatment and intense immunostaining after the treatment. CONCLUSIONS: These results confirm recent demonstrations of EGFRs in human testis and evidence different EGFR immunostaining in the presence of various degrees of testicular damage, suggesting a role of this growth factor in growth and differentiation of the germ cells throughout spermatogenesis. The observation that intense EGFR immunostaining was found in subjects showing high FSH plasma levels and in all of the patients who received exogenous FSH, supports a possible role of this gonadotropin in the modulation of the EGFR expression.

Erythropoietin stimulates testosterone production in man.

Human recombinant erythropoietin (rHuEPO) treatment improves sexual function in end-stage renal failure patients with a still-debated mechanism. Experimental data suggested that rHuEPO was able to stimulate rat Leydig steroidogenesis; therefore, it has been suggested that rHuEPO may induce its effects in humans by acting on gonadal steroid production. Thirteen young adult males (age range, 16-28 yr) catheterized at peripheral and left internal spermatic venous levels during a contrast study for varicocele, were studied. In five subjects, rHuEPO (60 IU/kg, up to a maximum of 4000 IU total) was injected over 1 min in the cubital vein. Similarly, in other five patients, 50 micrograms GnRH were infused. In three subjects, 2 mL saline were injected, as controls. Plasma LH, FSH, and testosterone (T) levels were then determined at -15, 0, 15, 30, 45, 60, 90, and 120 min simultaneously in peripheral and spermatic venous blood. rHuEPO infusion did not have any effect on plasma LH and FSH levels in peripheral or spermatic veins. Similarly, rHuEPO infusion did not affect peripheral T concentration, but increased (approximately 400% vs. controls; P < 0.05) spermatic T levels. GnRH infusion induced an increase in plasma LH and FSH levels in both peripheral and spermatic veins. After GnRH infusion, an increase of approximately 12-fold (P = 0.05-0.001) in T was observed only at the spermatic venous level, without any peripheral T variation. These findings show that rHuEPO was able to influence testicular steroidogenesis by stimulating T production in man, whereas the absence of any effect on gonadotropin secretion suggests that rHuEPO might act directly on human Leydig cell function.

Assessment of testicular cytology by fine needle aspiration as a diagnostic parameter in the evaluation of the oligospermic subject.

OBJECTIVE: To investigate whether testicular cytology by fine needle aspiration may be considered a diagnostic parameter in the evaluation of the oligospermic subject. DESIGN: Cytologic smears were obtained using a 23-gauge needle, stained with May-Grünwald-Giemsa stain and examined under a light Orthoplan microscope (Wild Leitz, Wetzlar, Germany) for qualitative and quantitative analysis. PATIENTS: One hundred sixty-six oligospermic patients were analyzed, and the findings were compared with those obtained from 40 normozoospermic infertile subjects used as controls. MAIN OUTCOME MEASURE(S): At least 200 spermatogenic cells were counted per slide and classified at the various steps of spermatogenesis. Because the number of Sertoli cells may be considered as a constant per unit of tubular length, Sertoli cells:spermatogenic cells ratio (termed Sertoli index) provided further elucidation and more comprehensible results. RESULTS: The procedure provided no sign of traumatization. The cytologic analysis permitted identification of different classes of oligospermic subjects, characterized by the following specific cytologic pictures: [1] bilateral or [2] unilateral germ depopulation (hypospermatogenesis), associated with maturation abnormalities of the first steps of spermatogenesis on both testes; [3] difficult maturation of the immature germ cells (spermatogonial or spermatocytic arrest); [4] ineffective spermiogenesis (spermatidic arrest); [5] normal maturation of the germ line in the presence of oligospermia, resulting from a transient acute damage on the spermatogenic line. Plasma levels of FSH and the testicular volumes agree with the cytologic picture of each group of patients. CONCLUSIONS: The results support use of fine needle aspiration of the testis as a minimally invasive diagnostic parameter for the assessment of oligospermic subjects.

Sperm nuclear instability and staining with aniline blue: abnormal persistence of histones in spermatozoa in infertile men.

During mammalian spermiogenesis, replacement of the somatic histones by basic proteins, the protamines, allows normal sperm nuclear condensation. In this study we have evaluated the degree of chromatin compaction in spermatozoa from 191 infertile subjects, affected by different testicular disorders, compare with that in 50 fertile sperm donors (controls). In infertile men, there was a higher percentage of unstable spermatozoa after incubation with sodium dodecyl sulphate (SDS) and of stained spermatozoa after staining with aniline blue (P less than 0.001 vs. controls). Furthermore, a positive linear correlation was found between SDS-unstable spermatozoa and stained spermatozoa (P less than 0.001), suggesting that sperm instability was related to a defect in histone-replacement by sperm-specific nucleoproteins, protamines. When the patients were considered according to pathology, high sperm nuclear instability and a high percentage of stained spermatozoa were detected in groups affected by varicocele, idiopathic infertility and in patients with a history of unilateral cryptorchidism. In the latter group the same alterations were observed even when the cryptorchid testis had been removed during surgery. In the group with a past history of mumps orchitis these parameters did not show any difference when compared with controls.

Assessment of testicular cytology by fine needle aspiration as a diagnostic parameter in the evaluation of the azoospermic subject.

OBJECTIVE: To investigate whether testicular cytology by fine needle aspiration (FNA) may be considered a diagnostic parameter in the evaluation of the azoospermic subject. DESIGN: Cytologic smears were obtained using a 23-G needle, stained with May-Grünwald-Giemsa stain and examined under a light Orthoplan microscope (Wild, Leitz, Germany) for qualitative and quantitative analysis. PATIENTS: Fifty-four azoospermic patients were analyzed, and the findings were compared with those obtained from 40 normozoospermic infertile subjects used as controls. MAIN OUTCOME MEASURE(S): Two hundred spermatogenic cells were counted and classified at the various steps of spermatogenesis. Spermatic index and Sertoli index provided further elucidations and more comprehensible results. RESULTS: No sign of traumatization was observed. Cytologic analysis was proved to have high statistical reproducibility (P less than 0.01 for spermatogonia and secondary spermatocytes and P less than 0.001 for the other cell types, when compared between differential counts) and permitted identification of different situations associated with azoospermia: Sertoli cell-only syndrome, germ depopulation (hypospermatogenesis), spermatogonial arrest, spermatidic arrest, and obstructive azoospermia. These findings agreed with clinical and hormonal parameters and with the results of bilateral surgical biopsies, when performed. CONCLUSIONS: The results support use of FNA of the testis as a noninvasive diagnostic parameter for the assessment of azoospermic subjects.

Evidence for the involvement of sperm angiotensin converting enzyme in fertilization.

Recently it has been observed that ejaculated human sperm possess high angiotensin converting enzyme (ACE) activity and that this enzyme is released during the process of capacitation. This observation raises the possibility that ACE may be involved in the fertilization process. To verify this hypothesis, we tested the effects of a potent ACE inhibitor, Captopril, on acrosome reaction induced by capacitating medium (3.5% HSA-added BWW) and on ability of human capacitated spermatozoa to penetrate zona-free hamster oocytes. Addition of Captopril (100 nmol l-1) modified neither sperm motility nor viability at any time considered, but significantly reduced the acrosome reaction percentages of sperm incubated in capacitating medium. Furthermore, Captopril significantly reduced the percentage of penetrated oocytes. The mean penetration rates both in the absence and presence of Captopril were 65.5 +/- 4.9% and 26.9 +/- 2.3% (P less than 0.001) respectively. These findings provide evidence that sperm release of ACE during capacitation may have a physiological role in the regulation of the mechanisms that allow sperm acrosome reaction and thus fertilizability.

Epidermal growth factor receptors (EGFR) localization in human testis.

An indirect immunofluorescence technique was used to detect and localize epidermal growth factor receptors (EGFR) in human testicular biopsies in different testicular diseases. Monolateral biopsies from twelve infertile subjects were studied from qualitative/quantitative points of view and were examined by immunofluorescence study with anti-EGFR monoclonal antibody. Two different patterns of EGFR expression were observed: a very weak presence of EGFR was detectable on Sertoli cells, peritubular basal structures, and interstitial compartment in testicular biopsies showing a normal spermatogenic status; and an important increase in EGFR expression was observed on the cytoplasm of Sertoli cells and on peritubular basal structures in biopsies exhibiting various degrees of hypospermatogenesis and in a case of Sertoli cells only syndrome. Germ cells did not show EGFR immunolocalization. EGFR seems to be present on different testicular target cells. EGFR expression increases on peritubular and Sertoli cells in the presence of significant tubular damage.

Stimulatory effects of alpha-hANP on testosterone secretion in man.

Several recent observations suggest that atrial natriuretic peptides (ANP) can modulate steroidogenesis in isolated rat Leydig cells. At present, it is unknown whether ANP influence human testicular steroidogenesis. We therefore evaluated the effects of alpha-human ANP (hANP) administration on testosterone plasma levels in peripheral and internal spermatic venous blood of young men (catheterized for contrast study of varicocele). Six subjects were injected with 100 micrograms alpha-hANP in the cubital vein. Six different patients similarly received 50 micrograms LHRH. Three controls received 2 ml saline. Plasma LH, FSH, and testosterone were then determined 15 min before, at time of injection, and 15, 30, 45, and 60 min thereafter in spermatic vein and peripheral venous blood, as well as at 120 min in peripheral blood. LHRH--induced LH increase was followed by a marked increase of spermatic vein testosterone concentrations, but the peripheral testosterone concentration did not increase. Similarly, alpha-hANP administration did not affect peripheral testosterone and LH concentrations, but significantly increased spermatic vein testosterone levels (P less than 0.01). Our findings demonstrate that alpha-hANP exerts its stimulatory effect on testicular steroidogenesis in man without modifying gonadotropin secretion, suggesting that alpha-hANP may directly influence Leydig cell function.

Immunomagnetic method to select human sperm without sperm surface-bound autoantibodies in male autoimmune infertility.

Using supermagnetic polymer microspheres coated with anti-immunoglobulins, spermatozoa without autoantibodies bound on their surface can be isolated from a sperm population showing a variable percentage of cells with autoantibodies bound on their surface. This simple technique seems to induce no modification of semen qualities; therefore, the concentration of sperm after immunomagnetic separation might be useful for in vivo or in vitro insemination in infertile couples with autoimmune male infertility.

Pharmacokinetics of miocamycin in patients with liver cirrhosis.

The pharmacokinetics of miocamycin, a new macrolide antibiotic, was studied in 6 healthy controls and 6 patients with compensated liver cirrhosis. The results indicate that in chronic liver disease the kinetics of the drug is altered. Therefore, a dosage adjustment of miocamycin is recommended when dealing with these patients.