A rapid and simple method for the separation of pure lymphocytes from horse blood

A method for the separation of pure and viable lymphocytes and granulocytes from the same blood sample in horses was reported. By centrifuging equine heparinized blood at 100 xg for 10 min at room temperature (r.t.), the resulting supernatant plasma was an almost pure (97.71 +/- 0.30%; n = 15) suspension of highly viable (98.72 +/- 0.28%) lymphocytes. When sodium citrate was used as an anticoagulant, lymphocyte suspensions collected in the same manner showed lower purity (87.89 +/- 1.59%; n = 9) and higher yields (56.56 +/- 3.89%, n = 9 versus 36.11 +/- 2.23%, n = 15). Where needed, a further centrifugation at 250 xg for 3 min (r.t.) of heparinized lymphocyte preparations removed an average of 87.39% (n = 15) contaminating platelets. A suspension of 85.96 +/- 2.20% pure granulocytes (93.23 +/- 1.74% neutrophils; n = 14) with minimal contamination by erythrocytes and high viability (93.11 +/- 1.26%) was obtained by performing a flash red blood cell lysis on the white-greyish layer resulting from the centrifugation of the heparinized blood samples. Among the several methods available, the procedure described herein is easy, rapid, cheap and reproducible.     

Renal diseases of reptiles.

Renal disease is one of the most common problems in reptilian medicine. Herbivorous species seem to be more susceptible than carnivores. The causation may be multifactorial, and the signs are often nonspecific. Prevention is preferred to treatment, and a comprehensive health program, which includes quarantine, routine health screens during annual examinations, and necropsy examinations of all animals that die, is desirable.     

A bio-economic indicators suite for the appraisal of the demersal trawl fishery in the Southern Adriatic Sea (Central Mediterranean)

The state of demersal fishery in the Southern Adriatic Sea (GFCM-GSA 18, Central Mediterranean), years 1996–2003, from a biological, social and economic point of view was analysed using 47 indicators: 22 biological indicators obtained from fishery-independent data through yearly experimental bottom trawl surveys (“Medits” Programme), and 25 socio-economic indicators estimated from fishery-dependent data, available from the monitoring system of the Italian Institute for Economic Research on Fisheries and Aquaculture (IREPA). Biological indicators were applied for “single-species” (Eledone cirrhosa, E. moschata, Illex coindetti, Merluccius merluccius, Mullus barbatus, Nephrops norvegicus, Parapenaeus longirostris, Raja clavata, Zeus faber) and for “multi-species” analysis. Economic indicators describing economic performance, productivity, costs and prices, and the overall economic sustainability of fishery were estimated. Social indicators and a general indicator summarising social sustainability were also considered. Indicators’ values were displayed using the Traffic Light system. Both fishery-independent and fishery-dependent indicators highlighted a progressive decline of the trawl fishery system in the GSA 18. This decline was mainly related to the ongoing depletion of the traditional fishery target species (mostly long-living, late-maturing species) partially replaced by the increase of traditional accessory species (generally short-living species), as well as to the reduction of productivity and increasing costs. The whole procedure was proposed as a contribution to the identification and applicability of bio-economic indicators for fishery management purposes.     

Prediction of blood orange MT firmness by multivariate modelling of low alterative penetrometric data set: A preliminary study

A study was conducted to evaluate a non-destructive method able to predict Magness–Taylor (MT) penetrometric firmness of blood oranges, potentially applicable for on-line selection. The method consisted of multivariate modelling of non-destructive penetrometric measurements performed on intact oranges with a 11.3 mm diameter probe by digital dynamometric equipment. The tests were carried out on 366 blood oranges (Citrus sinensis (L.) Osbeck cv. Tarocco); 60 were used as a control group to verify the non-destructive nature of the tests. Four test factors were used on the fruit: two different levels of non-destructive deformation (1 and 3 mm), at two different deformation speeds (5 and 30 mm s^?1). In order to obtain the reference parameter (MT firmness), a destructive penetrometric test followed each non-destructive deformation on the same fruit (deformation of 20 mm at 1 mm s^?1). The force–deformation curves of the non-destructive tests were elaborated to calculate different texture parameters (max force, elasticity, work). These parameters with raw curve data (force values for constant deformation steps) were used, after autoscaling pre-processing, as X-block datasets in supervised multivariate modelling (partial least squares) to predict MT firmness values. Among the test factors used, the 30 mm s^?1 speed and 3 mm deformation provided the best overall prediction of MT firmness. This comprised an r value of 0.76 and standard error of prevision (SEP) of 2.99 N for a MT firmness mean value of 35.65 N (8.4%). The time needed to apply the selected deformation may lead to a sorting speed of about 10 fruit per second, representing an acceptable value for commercial application.     

Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice and correction methods for quantitative assays of transgenic and endogenous GUS

Background  

The β-glucuronidase (GUS) gene reporter system is one of the most effective and employed techniques in the study of gene regulation in plant molecular biology. Improving protocols for GUS assays have rendered the original method described by Jefferson amenable to various requirements and conditions, but the serious limitation caused by inhibitors of the enzyme activity in plant tissues has thus far been underestimated.     

Food medicine and minor nourishment in the folk traditions of Central Italy (Marche,Abruzzo and Latium)

A series of research projects on wild plants used in Central Italy (Marche, Latium and Abruzzo) as food medicine and for minor nourishment was carried out in the years 1979-2000. This study reports a summary of the data personally collected in the field. Minor nourishment appears to be still consistently utilised: wild plants are consumed preferably in salads, boiled or in omelettes. Some therapeutic properties are attributed by people to the alimentary plants, often documented by the scientific literature. In this study, folk alimentary and/or therapeutic food uses of 126 entities distributed among 39 families are described. Particular folk uses concerning cultivated plants are also reported. In this paper, for the first time pharmacobotanical properties are systematically emphasised for the folk food species of Central Italy. References to analogous alimentary uses in Central Italy are documented. Comparisons among different food traditions are also discussed.     

Photoinhibition of photosynthesis in needles of two cypress (Cupressus sempervirens) clones

Photoinhibition of photosynthesis and photosynthetic recovery were studied in detached needles of cypress (Cupressus sempervirens L.) Clones 52 and 30 under controlled conditions of high irradiation (about 1900 micromol m(-2) s(-1) for 60 min; HL treatment), followed by 60 min in darkness. The degree of photoinhibition was determined based on the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm), which is a measure of the potential efficiency of photosystem II (PSII), and on electron transport measurements. The Fv/Fm ratio declined in needles of both clones in response to the HL treatment. Minimal fluorescence (Fo) increased in HL-treated needles of both clones. The HL treatment decreased rates of whole-chain and PSII activity of isolated thylakoids more in Clone 52 than in Clone 30. In needles of both clones, PSI activity was less sensitive to photoinhibition than PSII activity. In the subsequent 60-min dark incubation, fast recovery was observed in needles of both clones, with PSII efficiencies reaching similar values to those in non-photoinhibited needles. The artificial exogenous electron donors diphenyl carbazide (DPC), hydroxylamine (NH2OH) and manganese chloride (MnCl2) failed to restore the HL-induced loss of PSII activity in needles of Clone 30, whereas DPC and NH2OH significantly restored PSII activity in photoinhibited needles of Clone 52. Quantification of the PSII reaction center protein D1 and the 33-kDa protein of the water-splitting complex following HL treatment of needles revealed pronounced differences between Clone 52 and Clone 30. The large decrease in PSII activity in HL-treated needles was caused by the marked loss of D1 protein and 33-kDa protein in Clone 30 and Clone 52, respectively.     

Traditional phytotherapy in Central Italy (Marche, Abruzzo, and Latium)

In this study, the more significant results of extensive ethnopharmacobotanical research carried out by the author in the years 1977-2000 in 175 localities of three regions of Central Italy (Marche, Abruzzo, and Latium) have been reported and compared. The usages of 80 species belonging to 36 families are described, of which 71 were used in human therapy and 29 in veterinary medicine. Uses are suited with the number of localities in which they have been mentioned. Among the wild plant mainly still used, Malva sylvestris, Urtica dioica, and Sambucus nigra are particularly highly considered, while major uses of plants concern these plants in addition to Allium sativum, Rubus ulmifolius, Parietaria diffusa, Cynodon dactylon, and Ficus carica. Unusual phytotherapic uses concern Brassica oleracea, Taraxacum officinale (warts), Ruscus aculeatus, S. nigra (chilblains), Allium cepa (chilblains; to remove thorns and splinters), Juglans regia, R. ulmifolius (burns), and Euphorbia paralias (bites of weevers). Among new uses with only one quotation, we remember Cirsium arvense (intestinal disturbances), Centaurea bracteata (cough), Lupinus albus (calluses), Melittis melissophyllum (eye inflammations, antispasmodic), and Artemisia absinthium (tendon inflammations), while among plants employed in various regions with interesting less-known properties, there are C. arvense (emergency haemostatic), P. diffusa (insect bites), and Scrophularia canina (antiseptic and cicatrizing agent for wounds in bovines and sheep).     

Supercritical CO2 extract of Cinnamomum zeylanicum: chemical characterization and antityrosinase activity

The volatile oil of the bark of Cinnamomum zeylanicum was extracted by means of supercritical CO2 fluid extraction in different conditions of pressure and temperature. Its chemical composition was characterized by GC-MS analysis. Nineteen compounds, which in the supercritical extract represented >95% of the oil, were identified. (E)-Cinnamaldehyde (77.1%), (E)-beta-caryophyllene (6.0%), alpha-terpineol (4.4%), and eugenol (3.0%) were found to be the major constituents. The SFE oil of cinnamon was screened for its biological activity about the formation of melanin in vitro. The extract showed antityrosinase activity and was able to reduce the formation of insoluble flakes of melanin from tyrosine. The oil also delayed the browning effect in apple homogenate. (E)-Cinnamaldehyde and eugenol were found to be mainly responsible of this inhibition effect.