Impacts of short-term water restriction on Pelibuey sheep: physiological and blood parameters.

One of the projected effects of climate change is a reduction in rainfall in certain regions of the world. Hence, the agricultural and livestock sectors will have to cope with increasing incidences of water shortage while still maintaining productivity levels to feed an ever increasing global population. This short communication reports on the effect of a 2-week water stress on Pelibuey sheep in Cuba. Three treatments were compared, viz. supply of water ad libitum, water supplied once every 3 or 6 days. Following exposure to the water stress, the results showed no changes in sheep body weight or rectal temperature. However, respiration frequency was affected with water stress causing a reduction from 23.3 to 13.3 respirations per min in control and water-deprived animals, respectively. Furthermore, there was evidence for hemoconcentration in response to water stress (levels of hemoglobin increased from 9.2 to 13.1 g L-1 and hematocrits from 27.6 to 39.3% in the control group and animals restricted to water once every 6 days. The imposed water stress was also evident in the reduction of lymphocytes (from ±63 to 43%), and in increase of neutrophils (from approximately 38 to 54%) and leukocytes (from 3133 to 4933 per mm3). The results indicated a decline in the levels of antioxidants, i.e., SOD from approximately 13 to 10 U mg-1 protein and CAT activity from 23 to 9 U mg-1 protein. To the best of our knowledge, this is the first report on the response of Pelibuey sheep to short-term water shortage stress under Cuban environmental conditions.

Pineapple stem-derived bromelain based priming improves pepper seed protein reserve mobilization, germination, emergence and plant growth.

Pepper seeds are slow to germinate and emergence is often non-uniform and incomplete, reducing gains from this cash crop. This study investigated the effects of pineapple stem- derived protease (stem bromelain) based priming on pepper seed germination in relation to reserve mobilization (specifically, proteins and amino acids), germination, emergence and plant growth. These parameters were compared across two controls, (1) unsoaked seeds and (2) seeds soaked in deionized water, and seeds soaked in pineapple stem bromelain crude extract (treatment). Seeds were soaked in bromelain crude extract possessing a proteolytic activity of 6.25 tU or deionized water (first control) for 3 h at 35 °C. Light microscopy revealed an abundance of protein bodies in the endosperm of the seeds prior to imbibition. When observed for a period of 96 h, these bodies were progressively degraded, with the rate of this degradation being fastest in bromelain-treated seeds. Quantitative analysis of protein levels confirmed this observation: 17.2 mg proteins/g FW at 120 h after priming in bromelain-treated seeds compared with 22.1 mg/g FW in controls (average). The bromelain treatment also increased levels of free amino acids from 3.9 mg/g FW in the controls to 4.6 mg/g FW after 120 h of imbibition. Germination and emergence percentages were initially higher in bromelain-treated seeds: 92.0% germination in bromelain-treated seeds vs. ~ 52.2% in the controls at 18 d; 100% emergence in protease-treated seeds vs. ~ 72.2% in the controls at 18 d. However, these parameters were comparable across the treatment and the controls at 28 d. Importantly, plant fresh and dry weights were significantly higher when seeds were primed with bromelain. The results suggest the use of bromelain extracts for priming pepper seeds based on their proteolytic activity, since germination is dependent on the availability of crude protein and essential amino acids. The benefits of bromelain seed priming appear to translate into improved seedling growth as well.

Euclidean distance can identify the mannitol level that produces the most remarkable integral effect on sugarcane micropropagation in temporary immersion bioreactors.

Plant scientists usually record several indicators in their abiotic factor experiments. The common statistical management involves univariate analyses. Such analyses generally create a split picture of the effects of experimental treatments since each indicator is addressed independently. The Euclidean distance combined with the information of the control treatment could have potential as an integrating indicator. The Euclidean distance has demonstrated its usefulness in many scientific fields but, as far as we know, it has not yet been employed for plant experimental analyses. To exemplify the use of the Euclidean distance in this field, we performed an experiment focused on the effects of mannitol on sugarcane micropropagation in temporary immersion bioreactors. Five mannitol concentrations were compared: 0, 50, 100, 150 and 200 mM. As dependent variables we recorded shoot multiplication rate, fresh weight, and levels of aldehydes, chlorophylls, carotenoids and phenolics. The statistical protocol which we then carried out integrated all dependent variables to easily identify the mannitol concentration that produced the most remarkable integral effect. Results provided by the Euclidean distance demonstrate a gradually increasing distance from the control in function of increasing mannitol concentrations. 200 mM mannitol caused the most significant alteration of sugarcane biochemistry and physiology under the experimental conditions described here. This treatment showed the longest statistically significant Euclidean distance to the control treatment (2.38). In contrast, 50 and 100 mM mannitol showed the lowest Euclidean distances (0.61 and 0.84, respectively) and thus poor integrated effects of mannitol. The analysis shown here indicates that the use of the Euclidean distance can contribute to establishing a more integrated evaluation of the contrasting mannitol treatments.

IMPACT OF LIQUID NITROGEN EXPOSURE ON SELECTED BIOCHEMICAL AND STRUCTURAL PARAMETERS OF HYDRATED Phaseolus vulgaris L. SEEDS.

BACKGROUND: It is well known that cryopreserving seeds with high water content is detrimental to survival, but biochemical and structural parameters of cryostored hydrated common bean seeds have not been published. OBJECTIVE: The objective of this work was to study the effect of liquid nitrogen exposure on selected biochemical and structural parameters of hydrated Phaseolus vulgaris seeds. MATERIALS AND METHODS: We cryopreserved seeds at various moisture contents and evaluated: germination; electrolyte leakage; fresh seed weight; levels of chlorophyll pigments, malondialdehyde, other aldehydes, phenolics and proteins; thickness of cotyledon epidermis, parenchyma, and starch storage parenchyma; and radicle and plumule lengths. RESULTS: Germination was totally inhibited when seeds were immersed in water for 50 min (moisture content of 38%, FW basis) before cryopreservation. The combined effects of seed water imbibition and cryostorage decreased phenolics (free, cell wall-linked, total), chlorophyll a and protein content. By contrast, electrolyte leakage and levels of chlorophyll b and other aldehydes increased as a result of the combination of these two experimental factors. These were the most significant effects observed during exposure of humid seed to liquid nitrogen. CONCLUSION: Further studies are still required to clarify the molecular events taking place in plant cells during cryostorage.

Safety and efficacy of various concentrations of topical lidocaine gel for intravitreal injection.

INTRODUCTION: Intravitreal injection (IVT) is one of the most common vitreoretinal procedures, a large majority are performed with local anesthesia. The purpose of this study was to investigate the safety to the cornea and anesthetic efficacy of five concentrations of lidocaine gel. METHODS: A prospective clinical trial was conducted testing lidocaine gel in five preparations: 2, 3.5, 5, 8 and 12%. Patients with macular degeneration, diabetic edema or retina vein occlusion were scheduled for intravitreal treatment received topical anesthesia with lidocaine gel 5 and 10 min before the procedure. Patients answered the visual analog scale for pain during the procedure. Corneal and conjunctival was evaluated using the Oxford scale. RESULTS: In total, 260 patients were randomized into five groups. The mean pain scores (± standard deviation) were 2.63 (± 1.68) in the 2% group, 2.08 (± 1.35) in the 3.5%; 2.00 (± 1.65) in the 5%, 1.93 (± 1.40) in the 8% and 1.83 (± 1.35) in the 12% group. Mean pain score among all groups was similar (p = 0.077). There was no significant difference between groups in regard to keratitis mean score (p = 0.897). CONCLUSIONS: Lidocaine gel at concentrations from 2 to 12% induced similar anesthetic effect for IVTs, without adverse effects on cornea and conjunctiva.

Effects of cryopreservation of Phaseolus vulgaris L. seeds on early stages of germination.

In this work, we studied the effects of cryopreservation on various parameters of early stages of germination of Phaseolus vulgaris seeds (0, 7 and 14 days). Percentages of germination, fresh mass of different plant parts, levels of chlorophyll pigments (a, b, total), malondialdehyde, other aldehydes, phenolics (cell wall-linked, free, and total) and protein were determined. No phenotypic changes were observed visually in seedlings recovered from cryopreserved seeds. However, several significant effects of seed liquid nitrogen exposure were recorded at the biochemical level. There was a significant negative effect of cryopreservation on shoot protein content, which decreased from 3.11 mg g(-1) fresh weight for non-cryopreserved controls to 0.44 mg g(-1) fresh shoot weight for cryopreserved seeds. On the other hand, cryopreservation significantly increased levels of other aldehydes than malondialdehyde in shoots at day 7, from 56.47 µmol g(-1) for non-cryopreserved controls to 253.19 µmol g(-1) fresh shoot weight for cryopreserved samples. Liquid nitrogen exposure significantly reduced phenolics contents (free, cell-wall linked, total) in roots at day 7 after onset of germination. In general, roots were more affected by cryostorage compared with other plant parts, while leaves were the least affected. The effects of seed cryopreservation seem to decline progressively along with seedling growth.

Actividad proteolítica de extractos enzimáticos obtenidos de plantas de la familia Bromeliaceae / Proteolytic activity of enzymatic extracts from plants of the Bromeliaceae family

Las enzimas proteolíticas aisladas de plantas de la familia Bromeliaceae se utilizan ampliamente en la industria médica, biotecnológica y alimenticia. Los estudios realizados en los últimos años sobre la actividad contra metástasis y tumores de las cisteíno-proteasas hacen que se incremente el interés por explorar nuevas fuentes naturales de obtención de fitoproteasas. En el presente trabajo se evaluó la actividad proteolítica de extractos enzimáticos obtenidos a partir de diferentes órganos de plantas de la familia Bromeliaceae. Se colectaron y clasificaron cinco grupos. Las plantas que se colectaron pertenecen a 3 géneros de la mencionada familia: 3 grupos son del género Tillandsia, 1 es del género Guzmania y otro del género Hohenbergia. Los mayores índices de actividad específica (3,3 U/mg de proteínas) se obtuvieron en los preparados obtenidos a partir de diferentes órganos de Hohenbergia penduliflora Mez, de cuyos extractos obtenidos se evaluó la influencia del pH de extracción y la actividad específica fue superior al realizarla a pH 3 a partir de sus tallos(AU)

Actividad proteolítica de extractos enzimáticos obtenidos de plantas de la familia Bromeliaceae / Proteolytic activity of enzymatic extracts from plants of the Bromeliaceae family

Las enzimas proteolíticas aisladas de plantas de la familia Bromeliaceae se utilizan ampliamente en la industria médica, biotecnológica y alimenticia. Los estudios realizados en los últimos años sobre la actividad contra metástasis y tumores de las cisteíno-proteasas hacen que se incremente el interés por explorar nuevas fuentes naturales de obtención de fitoproteasas. En el presente trabajo se evaluó la actividad proteolítica de extractos enzimáticos obtenidos a partir de diferentes órganos de plantas de la familia Bromeliaceae. Se colectaron y clasificaron cinco grupos. Las plantas que se colectaron pertenecen a 3 géneros de la mencionada familia: 3 grupos son del género Tillandsia, 1 es del género Guzmania y otro del género Hohenbergia. Los mayores índices de actividad específica (3,3 U/mg de proteínas) se obtuvieron en los preparados obtenidos a partir de diferentes órganos de Hohenbergia penduliflora Mez, de cuyos extractos obtenidos se evaluó la influencia del pH de extracción y la actividad específica fue superior al realizarla a pH 3 a partir de sus tallos.

The proteolytic enzymes isolated from the Bromeliaceae family are widely used in the medical, biotechnological, and food industries. The studies conducted in recent years on the activity against metastasis and cysteine-proteases, increase the interest in screening new natural sources of obtention of phytoproteases. In the present paper, the authors assessed the proteolytic activity of enzymatic extracts obtained from different organs of the Bromeliaceae family plants. Five groups were collected and classified. Plants obtained belong to three genuses of the above mentioned family: 3 groups are of genus Tillandsia , one of genus Guzmania , and the other of genus Hohenbergia . The highest rates of specific activity (3.3 U/mg of proteins) were attained in preparations obtained from different organs of Hohenbergia penduliflora Mez. from whose extracts the influence of extraction pH was assessed. The specific activity was greater on carrying it out at pH3, starting from their stalks.