Age-related ultrastructural and monoamine oxidase changes in the rat optic nerve.

The aim of this paper is to study the morphology and the distribution of the monoamine oxidase enzymatic system in the optic nerve of 4 month-old Wistar (young) and 28 month-old Wistar (old) rats. The optic nerve was harvested from 20 young and old rats. The segment of optic nerve was divided longitudinally into two pieces, each 0.1 mm in length. The first piece was used for transmission electron microscopy. The second piece was stained with histochemical reaction for monoamine oxidase. The agerelated changes in the optic nerve of rats include micro-anatomical details, ultrastructure and monoamine oxidase histochemical staining. A strong decrease of the thin nerve fibers and a swelling of the thick ones can be observed in optic nerve fibers of old rats. Increased monoamine oxidase histochemical staining of the optic nerve of aged rats is well demonstrated. The increase of meningeal shealth and the decrease of thin nerve fibers of the optic nerve in old rats are well documented. Morphological, ultrastructural and histochemical changes observed in optic nerve fibers of the old rats show a close relation with aging.

Cryoprotectant-free vitrification of human spermatozoa in new artificial seminal fluid.

Vitrification is a new method that has been recently introduced in Assisted Reproduction Technique programs. The aim of this study was to design a new medium similar to normal human seminal fluid (SF), formulation artificial seminal fluid (ASF), and to compare the cryoprotective potency of this medium with SF and human tubal fluid (HTF) medium. Thirty normal ejaculates were processed with the swim-up technique and sperm suspensions were divided into four aliquots: (i) fresh sample (control); (ii) vitrification in HTF medium supplemented with 5 mg/mL human serum albumin and 0.25 mol sucrose (Vit HTF); (iii) vitrification with patients' SF (Vit SF); and (iv) vitrification in ASF (Vit ASF). After warming, sperm parameters of motility, viability, and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% glutaraldehyde and processed for scanning electron microscopy and transmission electron microscopy observations. The results showed that progressive motility (46.09 ± 10.33 vs. 36.80 ± 13.75), grade A motility (36.59 ± 11.40 vs. 16.41 ± 11.24), and normal morphology (18.74 ± 8.35 vs. 11.85 ± 5.84) and viability (68.22 ± 10.83 vs. 60.86 ± 11.72) of spermatozoa were significantly higher in Vit ASF than in Vit HTF. All parameters were better in Vit ASF than in Vit SF, but only viability was significantly different (p = 0.006). After cryopreservation, deep invagination in cytoplasm and mechanically weak point sites and folded tail were commonly observed. But, this phenomenon was more significant in Vit HTF and Vit SF than in ASF (p < 0.05). In transmission electron microscopy evaluation, acrosome damage, plasma membrane loss, chromatin vacuolation, and disruption of mitochondria arrangement and structures were observed in all vitrified groups. Adherence of several tail sections together was also seen in all cryo groups. But this was seen more in Vit HTF and Vit SF than in ASF (p < 0.05). In conclusion, vitrification of human spermatozoa with ASF can effectively preserve the quality of sperm motility in comparison with Vit HTF.

[Surveillance of health status in an agrarian environment: proposal for a method. 1. Epidemiological data]. / La sorveglianza dello stato di salute in ambiente agricolo: proposta di un metodo. Nota I--Dati epidemiologici.

In order to evaluate the risk of the exposure of the pesticides and N-nitroso compounds in the agricultural environment not only farm-hands have to be considered but also their families. So it is necessary to prepare survey methodologies to examine the state of health of all individuals who are exposed to same kind of risk (pesticides and nitroso compounds), even if in different ways and intensity. We have therefore chosen four farms situated on the same territory which cultivate vegetables. Three, of these farms, made wide use of pesticides and fertilizers, the fourth one was used as a "control farm" because it did not employ any chemicals. For every farm data about the neighbouring territory, the climate, the kind of cultivation, the pesticides and other employed chemicals were collected. Every subject (in all 25) residing in the farm was interviewed with a standard questionnaire about personal data, the duties performed on the farm, the way in which chemicals were used, their medical history as well as the life style. Samples of blood were drawn periodically in different seasons, over a two year period, to determine both nitrosoamine (NA) and the enzymes which reveal hepatic damage: Alkaline phosphatase (ALP), Leucinoaminopeptidase (LAP), Gamma-glutamyltransferase (GGT) and Acetylcholinesterase (Ach). We have observed that: a. the most frequent pathologies concern skin (16% of the people) and liver (12% of the people), which are favourite targets for agricultural chemicals though the serum enzymes did not show any important change; b. 8% of the subjects had acute pesticide poisoning; we therefore observed neither particular precautionary safety measures, nor a particular knowledge of acute and chronic toxic effects depending on the use of pesticides. So we suggest: 1) that the Sanitary Authorities control the state of health not only of the farm-hands but also of their families, if exposed to risk; this can be realized through the strict collaboration of the Occupational Physician and the Family doctor; 2) capillary action of health education regarding agricultural risks; 3) to increase research to find more sensitive and efficient biological indicators in order to evaluate the hazards of agricultural chemicals.