Epigallocatechin-3-gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C.

Stallion semen storage for artificial insemination is mainly based on liquid cooled storage. In many stallions this technique maintains sperm quality for an extended period of time (24-72 hr) at 7°C. While this technique is commonly used in the horse industry, there can be a decline in fertility in some stallions, due to an inability of their sperm to tolerate the cool storage process. The aim of the present work was to evaluate the effect of two natural antioxidants (epigallocatechin-3-gallate (EGCG) at 20, 60 and 120 µm and green tea polyphenols, and p at .001, .01 and .1 mg/ml) on some sperm parameters (sperm motility, viability/acrosome integrity and DNA quality) in extended semen immediately after its collection (T0) and after 2, 6, 24 and 48 hr of cool storage. Two ejaculates from three trotter stallions were analysed after 48 hr of storage at 4°C. No beneficial effect on the analysed parameters was observed: the two antioxidants were not able to improve sperm quality after 48 hr of storage. These results are in agreement with previous findings on the effect of different antioxidants reported by other researches, who have demonstrated that stallion semen keeps good antioxidant capacity after dilution for 24 hr. In conclusion, the positive effect exerted by antioxidant molecules in other species is not confirmed in the equine one.

Effect of cushioned or single layer semen centrifugation before sex sorting on frozen stallion semen quality.

The aim of this study was to compare the effect of presorting centrifugation (cushioned [CC] or single-layer colloid [SLC]), with simple dilution (SD), on the quality of sex-sorted stallion semen before and after sorting and after freezing and thawing. Four ejaculates from each of two fertile stallions were collected 1 week apart and evaluated for percent total sperm motility (TM), percent viable acrosome-intact sperm (VAI), and DNA quality (percentage of DNA fragmentation index). Freezing caused, independently from CC and SLC treatments, a significant decrease of TM (P < 0.05) and VAI (P < 0.05) in both unsorted and sorted semen. On the other hand, sorting did not impair TM and VAI and, interestingly, improved DNA quality in all treatments only before freezing (28 vs 13, 28 vs 10, 22 vs 7 in SD, CC, and SLC for unsorted vs sorted groups, respectively; P < 0.05); this positive effect was lost in the same samples after freezing and thawing, suggesting that the freezing process reduces the DNA quality of sex-sorted sperm. Our results suggest that CC and SLC are not able to select those spermatozoa that possess a better ability to withstand sperm processing associated with sperm sorting and freezing.

Effects of repeated transvaginal ultrasound-guided aspirations performed in anestrous and cyclic mares on P4 and E2 plasma levels and luteal function.

The aim of the present study was to verify how repeated ovum pick-up (OPU), performed in anestrous and cyclic mares, affect ovarian activity, measured by progesterone (P4) and 17ß-estradiol (E2) plasma levels. Ovum pick-up of all visible follicles was performed every 9 to 12 days, and four sessions were carried out during anestrous (A) and breeding season (BS). The number of aspirated follicles per mare at each session was not significantly different between the two periods (BS: 6.1 ± 2.4; A: 7.5 ± 4.4; P > 0.05), but the mean follicular diameter was significantly higher during BS (16.0 ± 7.1 vs. 10.2 ± 5.1 mm; P < 0.05); during A the number of aspirated follicles less than 15 mm in diameter resulted significantly higher than that registered in BS (5.1 ± 2.7 vs. 3.0 ± 1.8; P < 0.05). The total mean value of P4 was higher in BS than in A (6.3 ± 4.4 vs. 0.3 ± 1.8 ng/mL; P < 0.05), whereas the total mean level of E2 was not different between the two periods (3.8 ± 3.4 vs. 2.5 ± 2.7 pg/mL; P > 0.05). Estradiol plasma values resulted positively correlated, in A and BS, with diameter of follicles detected on the ovaries (R = 0.345 and R = 0.331, respectively), whereas a negative correlation was observed between P4 and follicular diameter in BS (R = -0.162). Both E2 and P4 presented a high individual variability during BS; in particular, in three of seven mares, P4 trend was compatible with a normal estrous cycle, and the interval between two consecutive peaks was 21 days. In two of seven mares, with CL at first OPU, P4 concentrations remained more than 3 ng/mL throughout the entire treatment period. Finally, in two of seven animals, P4 levels initially showed a similar pattern to that of a normal estrous cycle, then, after the second aspiration, they remained consistently higher than 3 ng/mL. When the procedure was carried out in cyclic animals, the influence of this technique on ovarian activity seemed to be related to individual variability although, according to progesterone values, structures observed on the ovaries after aspirations presented luteal function. Furthermore, the resumption of normal ovarian activity, after repeated OPU sessions, occurred in a period not much longer than the duration of a normal estrous cycle (25.4 ± 5.2 days). Data recorded during nonbreeding period showed that repeated OPU in anestrous mares do not affect ovarian activity and do not anticipate the resumption of ovarian cyclicity. However, based on the number of aspirated follicles in anestrous and cyclic mares, both types of subjects could be considered as oocyte donors.

Defining the clinical outcome status (COS) in sarcoidosis: results of WASOG Task Force.

The clinical outcome of sarcoidosis is quite variable. Several scoring systems have been used to assess the level of disease and clinical outcome. The definition of clinical phenotypes has become an important goal as genetic studies have identified distinct genotypes associated with different clinical phenotypes. In addition, treatment strategies have been developed for patients with resolving versus non resolving disease. A task force was established by the World Association of Sarcoidosis and Other Granulomatous diseases (WASOG) to define clinical phenotypes of the disease based on the clinical outcome status (COS). The committee chose to examine patients five years after diagnosis to determine the COS. Several features of the disease were incorporated into the final nine categories of the disease. These included the current or past need for systemic therapy, the resolution of the disease, and current status of the condition. Sarcoidosis patients who were African American or older were likely to have a higher COS, indicating more chronic disease. The COS may be useful in future studies of sarcoidosis.

Quality and fertilizing ability in vivo of sex-sorted stallion spermatozoa.

Little information is available on the quality of stallion spermatozoa after sex sorting. The objectives of the present study were to assess the quality of sex-sorted stallion spermatozoa and determine its fertilizing ability after hysteroscopic low dose insemination. Ejaculates from four stallions were collected and sorted by a MoFlo SX flow cytometer/sperm sorter. Before and after sorting, spermatozoa were evaluated for motility by Computer Assisted Sperm Analysis, viability (SYBR 14-propidium iodide), mitochondrial function (JC-1) and acrosomal status (fluorescein isothiocyanate Pisum sativum agglutinin conjugated). A fertility trial was carried out on four mares (seven oestrous cycles) by hysteroscopic insemination, depositing 5 x 10(6) X-bearing spermatozoa. Sex sorting resulted in a significant decrease (p < 0.001) in all motility characteristics. Sperm viability and percentage of spermatozoa with functional mitochondria were not affected by the sorting process, while the percentage of reacted spermatozoa was higher (p < 0.01) for non-sorted than sorted spermatozoa. Pregnancy rate was 28.6% (2/7) after low dose hysteroscopic insemination. Only one pregnancy was carried to term with the birth of a healthy filly. In conclusion, despite the reduction in sperm motility, sex sorting did not impair stallion sperm viability and mitochondrial activity immediately post-thaw; moreover, the sexed spermatozoa retained the ability to fertilize in vivo.

[Adoptive parental couple: a pilot study]. / La coppia genitoriale adottiva: uno studio pilota.

International adoption is a phenomenon in constant growth, involving over one hundred countries. In Italy there are approximately two thousand adoptions of foreign minors per year. The needs demonstrated by the families adopting these children, sometimes bearing complex problems, have led to the offer of a combined medical and psychological intervention oriented towards a global and integrated approach to the needs of a family that is changing with the arrival of a new member. Therefore, a clinical evaluation of the health status of the child recently arrived in Italy, targeted at identifying the presence of medical or psychological conditions, is performed during the day spent at the day hospital. The authors present data and considerations emerging from their ongoing experience, which has already involved 113 children and 89 families. The majority of the children evaluated so far show acute physical conditions, requiring major attention and support from their parents. The authors believe that in order to organize an adequate intervention it is useful to consider the parents of these children as caregivers. The function of caregiver is identified at three levels: the care of the child as son/daughter (typical of the parental function), the care of the problems deriving from the specific preadoption experience that accompany the child in the new family, and the care of the sick child. The authors deem that the combined medical and psychological intervention adopted at present with these families constitutes a useful support to the caregiver function.

Extrapulmonary presentation of sarcoidosis.

Sarcoidosis is a systemic disease that usually has a pulmonary presentation. Extrapulmonary presentation of the disease is often unrecognized because the clinical signs ( eg, peripheral lymph nodes) and more important symptoms ( eg, renal stones or uveitis) do not usually indicate systemic disease. In three different long-term studies of patients with sarcoidosis, 1.5% of the patients presented with uveitis, 11.7% presented with peripheral lymph nodes, and 3.6% presented with renal stones; a total of 16.6% of the patients had extrapulmonary presentation of sarcoidosis. Two of the three studies were retrospective. Many patients may have skin, parotid gland, spleen, liver, central nervous system, bone, or heart presentation; 16.6% is only the tip of an iceberg. A well-conducted, prospective, long-term study could indicate that more than 30% of patients with sarcoidosis have extrapulmonary presentations of the disease.

Comparison of registries of interstitial lung diseases in three European countries.

Few published studies have compared epidemiological data on the prevalence, incidence or relative frequency of the different interstitial lung diseases. In this review, the data of such registries from three countries in Europe (Belgium, Germany and Italy) are compared with those in the USA (Bernalillo County, NM). These registries show some striking similarities, but also discrepancies, which in part may be real, but may also be due to selection bias. Indeed, the registries in the European countries encompass the patients seen by pulmonologists, while the registry in Bernalillo County is based on a general population study in a limited area. In addition, in some studies, prevalences and incidences were registered, while in others, only prevalences or incidences were registered. Finally, most, but not all studies were prospective. Nonetheless, the different studies showed that sarcoidosis, idiopathic pulmonary fibrosis, hypersensitivity pneumonitis, interstitial lung diseases due to collagen vascular diseases and not-defined (or postinflammatory) fibrosis were most frequent. In the majority of disease entities a male predominance was found. Surprisingly large differences in the use of diagnostic techniques, such as high-resolution computed tomography, bronchoalveolar lavage, open lung biopsy and transbronchial lung biopsy, were noted between the different studies.

Interstitial lung diseases: an epidemiological overview.

Epidemiological studies on interstitial lung diseases (ILDs) may be schematically subdivided into the following major types: 1) quantifications of disease, broken down into incidence, prevalence and mortality data; 2) identification of aetiological factors; and 3) clinical epidemiological studies. Epidemiological data may be obtained from different sources or population groups, using different study designs such as systematic national statistics, population-based data and registries, and large case series of specific diseases. Differences in results between epidemiological studies may be due to real differences in incidence, but may also be due to changes in disease definitions and classifications, differences in the epidemiological design of the studies, or even registration bias. Comparative epidemiological data of different ILDs are almost limited to the general population study in Bernalillo County and to national mortality statistics, which should be interpreted with great caution. Also, some, mostly national registries of the different ILDs have been carried out by specific medical profession groups (especially pulmonologists), which clearly underestimate the real incidence of ILDs, but in which the comparison of the relative frequencies is probably accurate. Based on all these comparative studies, sarcoidosis and idiopathic pulmonary fibrosis appear to be the most frequent ILDs, followed by hypersensitivity pneumonitis and ILD in collagen vascular disease, when classical pneumoconioses are not included. There is also a relatively large group of nonspecific fibrosis. Much more data have been published on the epidemiology of specific forms of interstitial lung disease. Most information is available on the epidemiology of sarcoidosis, and those data are probably the most accurate. Data on idiopathic pulmonary fibrosis have the disadvantage of the recent changes in definition and classification of this disease. Hypersensitivity pneumonitis has been studied epidemiologically, especially in some exposure groups such as farmers and pigeon breeders, and in some regions in North America, UK, France and Scandinavia. Estimates of frequencies of interstitial lung disease in collagen vascular disease or of drug-induced interstitial lung disease are less accurate and more variable, depending on diagnostic criteria. Notwithstanding the aforementioned problems, this report tries to provide a balanced overview of the epidemiology of different interstitial lung diseases.

First report of the Italian register for diffuse infiltrative lung disorders (RIPID).

RIPID was established in 1998 as a joint project of the major Italian scientific societies for Respiratory Medicine, with the aim to create an Italian Register on diffuse infiltrative lung disorders that can provide the basis for epidemiological and clinical studies of adequate sample size. In the period from May 1998 to December 2000, 1,382 cases were submitted from 54 Centers in 15 regions of Italy, 54.2% males (mean age +/- SD 50.5 +/- 16.8 years) and 45.8% females (50.2 +/- 15.3 years). A current smoking habit emerges in 18% of subjects; former smokers and never-smokers represent 26% and 56% of the total case series, respectively. The most frequent disease registered is idiopathic pulmonary fibrosis (37.6%), followed in decreasing order by sarcoidosis (29.2%), and Langherans' cell hystiocytosis (6.6%). High resolution computed tomography (HRCT) was considered as the most important tool for final diagnosis in the majority of cases (74.4%); 39.4% of patients underwent transbronchial biopsies, 39.2% bronchoalveolar lavage (BAL). A surgical biopsy was performed in 20.5% of patients. A web site has been activated from December 2000 (www.pneumonet.it/ripid), allowing prompt access to all information and scientific material concerning the project and to an electronic form for data collection that can be completed on-line.

The clinical spectrum of the sarcoid peripheral lymph node.

BACKGROUND: The finding of sarcoid-type granulomas in a peripheral lymph node (PLN) without clinical evidence of changes suggestive of sarcoidosis elsewhere poses a diagnostic problem. The long term follow-up of these patients has never been described in adults. AIMS OF THE WORK: 1. To describe in the above population whether and when a definite diagnosis of sarcoidosis was eventually made, and the time required to make the diagnosis. 2. To study the percentage of peripheral lymph node presentation in sarcoidosis. PATIENTS: A peripheral lymph node presentation, with lymph node biopsy demonstrating sarcoid granulomas, was seen in 127 patients over the last 20 years. Detailed investigation permitted the early diagnosis of sarcoidosis in 76, and of sarcoid reaction in 8 patients. The other 43-patients with granulomatous lymph node and no clinical evidence of changes outside the lymphatic system at the onset are the subject of the present study. METHODS: Periodic examination at our Sarcoid Clinic every 2 to 4 months, in a long term median follow-up of 36 months (range 1 to 203) and workup according to clinical need, including chest X ray and Computed Axial Tomography (CAT), pulmonary function tests, total body 67Ga scan, Broncho Alveolar Lavage (BAL) studies, blood cell counts, 24 h calciuria and urine analysis, serological tests for liver function, calcaemia, Angiotensin Converting Enzyme (ACE). RESULTS: The diagnosis of sarcoidosis (chronic in all) could be made in 33 patients (25 pulmonary, 8 extrapulmonary), after a median time from presentation of 5 years (range 3-288 months). In the other 10, in spite of a median duration of the illness of 62 months (range 20-487), our diagnosis has been idiopathic granulomatous disease of peripheral lymph nodes. Thus, we observed 109 patients in 20 years presenting with lymph nodes that were surgically removed and provided the diagnosis of sarcoidosis sooner (76 patients) or later (33 patients). CONCLUSIONS: 1. In patients presenting only sarcoid granulomas in peripheral lymph nodes, sarcoidosis may be diagnosed months or years later, but a subpopulation of them still exists where granulomatous lesions remain unexplained. 2. In our series of patients, peripheral lymph node presentation occurred in 11.7% of cases of sarcoidosis.

The role of thoracic surgery in diagnosing interstitial lung disease.

In this article, the current status of thoracic surgery options for reaching a diagnosis in interstitial lung disease is described. When surgery is needed, mediastinoscopy is the first step in cases of suspected stage I or II sarcoidosis. If this is not the case, video-assisted thoracoscopy is currently preferred to open lung biopsy because the need for analgesia lessened, less blood is lost, the operative time is shorter, the complication rate is lower, and the postoperative stay is shorter. In some cases, video-assisted thoracoscopy may also be preferred to mediastinoscopy, especially in young women, for cosmetic reasons.

Changing strategies of lung biopsies in diffuse lung diseases: the impact of video-assisted thoracoscopy.

The aims of this report were: 1) to compare the strategy of bioptic approach in Italy during the last 4 yrs with a previous period; and 2) to compare efficacy and safety of video-assisted thorascopic lung biopsy (VTLB) versus OLB. We retrospectively evaluated: 1) the strategy of the bioptic approach in the Milan Sarcoid Clinic in the years 1992-1995 (201 patients) versus 1988-1991 (197 patients); and 2) data from 65 VTLB procedures in the years 1992-1995 versus 68 OLB procedures in the years 1988-1991 performed in patients with diffuse lung disease. It was found that the use of OLB (17-9%), mediastinoscopy (15-5%), and scalene node biopsy (20-7%) decreased, whereas transbronchial biopsy (TBB) increased (11-17%). VTLB biopsy is now performed in 17% of patients. VTLB compares favourably with OLB as there is less need for analgesia (7.5+/-7.5 versus 17.5+/-8.0 methadone mg i.m.: p<0.001), lower blood loss (61+/-58 versus 156+/-84 mL in the first postoperative day: p<0.001), and shorter postoperative stay (4.7+/-1.6 versus 5.7+/-1.4 days: p<0.001). Specimen adequacy (98.6 versus 985%) and diagnostic accuracy (86.1% VTLB, versus 92.6% OLB: p>0.05) were the same in the two groups. In conclusion, video-assisted thoracoscopic lung biopsy is replacing both mediastinoscopy and open lung biopsy. It is at present the best option when a surgical procedure is required for histological confirmation of diffuse lung disease.

The late follow-up of chronic sarcoid patients previously treated with corticosteroids.

AIM OF THE STUDY: The aim of the study was to evaluate, in a white population with chronic sarcoidosis, the rate and pattern of relapses, the correlated factors, and the course of disease after prednisone withdrawal. METHODS: We have retrospectively examined the charts of 702 consecutive patients with histologically proven sarcoidosis, first seen in the Milan Sarcoidosis Clinic in the period October 1978-October 1994. 239 patients required corticosteroid therapy; in 82 it was possible to discontinue prednisone therapy and to have a follow-up of at least 18 months after withdrawal. RESULTS: A relapse, requiring a new course of steroids, was observed in 30 (36.6%) of the 82 patients (R group). The other 52 patients (No-R group) did not relapse during a mean follow-up of 36.8 +/- 24.8 months (range 18-125). There were no relapses after 3 asymptomatic years of prednisone withdrawal. Extrapulmonary sarcoidosis was a reason for giving therapy in 46.6% of patients in the R group, vs 23.0% in the No-R group (P < 0.05). The first course of therapy lasted 22 months [median time; i.q. 11.5 to 34.5] in R group vs 26 months [i.q. 18 to 41] in No-R group (P > 0.05). The mean daily prednisone dose was higher in the R group: 17 mg [median value; i.q. 8.9 to 23.2] vs 10.6 mg [i.q. 8.1 to 13.8] in the No-R group (p < 0.05). Logistic regression confirmed the prognostic significance of mean daily prednisone dose and of extrapulmonary sarcoidosis at presentation (P < 0.01). A mild sarcoid activity at the time of withdrawal was still present in 51.9% of patients who did not relapse, and in 66.7% of patients who relapsed (p > 0.05). Relapse in the first year after withdrawal of prednisone therapy occurred in twenty-five of the 30 patients. The pattern of relapse was different from the initial manifestation in 5. Nine of the 30 patients could ultimately be weaned successfully from prednisone. CONCLUSION: Relapses occurred in 36.6% of cases, and their pattern was the same as the initial manifestation in the majority of cases. A mild sarcoid activity at the time of withdrawal is not a reason for continuing steroids when the disease is abating. In our white population severe irreversible pulmonary impairment is rare, and even patients requiring chronic therapy need low prednisone dosage, usually around 10 mg daily, to control the disease in the late course.