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Identification of a grapefruit cDNA belonging to a unique class of citrus dehydrins and characterization of its expression patterns under temperature stress conditions.

Citrus fruits are sensitive to low temperatures and this often results in the development of chilling injuries during postharvest storage. In order to gain more insight into the molecular mechanisms involved in the acquisition of fruit chilling tolerance, we initiated a grapefruit (Citrus paradisi, cv. Marsh Seedless) flavedo cDNA sequencing project and used it to identify a cDNA similar to other Poncirus trifoliata and Citrus unshiu dehydrin genes reported to be responsive to low temperatures. The grapefruit dehydrin cDNA, designated cor15, encodes a predicted polypeptide of 15.1 kDa, that is almost completely identical with other reported citrus dehydrin proteins, except that it contains two large amino acid repeats, whereas P. trifoliata COR11 has only one such repeat and P. trifoliata COR19 and C. unshiu COR19 have three repeats. Together, the various grapefruit, P. trifoliata and C. unshiu dehydrins form a closely related and unique dehydrin gene family that differs from most other plant dehydrins in having an unusual K-segment similar to that of gymnosperms and in having a serine cluster (S-segment) at an unusual position at the carboxy-terminus. The grapefruit cor15 gene is consistently expressed in the fruit peel tissue at harvest, but its message levels dramatically decrease during storage at 2 degrees C. However, a pre-storage hot water treatment, which enhances fruit chilling tolerance, elicited retention of the constant level of cor15 gene expression during cold storage and eliminated its decline. The hot water treatment had no inductive effect on cor15 gene expression when the fruit were held at non-chilling temperatures. The effects of other stresses, such as exposure to ethylene, UV irradiation and wounding, on cor15 gene expression, were temporary and persisted for 1-2 days after the treatments.

Journal Article↗

Deep-sea ecology. Developmental arrest in vent worm embryos.

Temperature is a key factor in controlling the distribution of marine organisms and is particularly important at hydrothermal vents, where steep thermal gradients are present over a scale of centimetres. The thermophilic worm Alvinella pompejana, which is found at the vents of the East Pacific Rise (2,500-m depth), has an unusually broad thermotolerance (20-80 degrees C) as an adult, but we show here that the temperature range required by the developing embryo is very different from that tolerated by adults. Our results indicate that early embryos may disperse through cold abyssal water in a state of developmental arrest, completing their development only when they encounter water that is warm enough for their growth and survival.

Animals↗

Prevention of natural colds by contact prophylaxis with intranasal alpha 2-interferon.

We conducted a randomized, placebo-controlled, double-blind study to determine whether intranasal alpha 2-interferon could prevent respiratory illnesses in healthy contacts of ill family members. Beginning within 48 hours of the onset of illness in a family member, contacts self-administered interferon (5 X 10(6) IU) or placebo spray once daily for seven days. Respiratory illness developed during the eight-day period, starting with the second day of spraying, in 52 of 222 persons in the placebo group as compared with 32 of 226 in the interferon group (P = 0.02; efficacy, 39 percent). Among persons exposed to laboratory-documented rhinovirus colds, illness developed in 2 of 27 interferon recipients as compared with 12 of 34 placebo recipients (P = 0.02; efficacy, 79 percent). During the two-week period during and after spraying, rhinovirus colds developed in 1.3 percent of those spraying with interferon and in 15.1 percent of those spraying with placebo (P = 0.003; efficacy, 88 percent). Blood-tinged mucus or nasal mucosal bleeding or both were detected in 7.7 percent of placebo and 13.6 percent of interferon users (P = 0.04), but no evidence of cumulative nasal toxicity was found. We conclude that postexposure prophylaxis with intranasal interferon may in some cases provide an effective strategy for controlling the spread of natural colds, especially those caused by rhinoviruses.

Administration, Intranasal↗

Echinacea purpurea for prevention of experimental rhinovirus colds.

A randomized, double-blind, placebo-controlled clinical trial was conducted to evaluate the ability of Echinacea purpurea to prevent infection with rhinovirus type 39 (RV-39). Forty-eight previously healthy adults received echinacea or placebo, 2.5 mL 3 times per day, for 7 days before and 7 days after intranasal inoculation with RV-39. Symptoms were assessed to evaluate clinical illness. Viral culture and serologic studies were performed to evaluate the presence of rhinovirus infection. A total of 92% of echinacea recipients and 95% of placebo recipients were infected. Colds developed in 58% of echinacea recipients and 82% of placebo recipients (P=.114, by Fisher's exact test). Administration of echinacea before and after exposure to rhinovirus did not decrease the rate of infection; however, because of the small sample size, statistical hypothesis testing had relatively poor power to detect statistically significant differences in the frequency and severity of illness.

Adolescent↗

New dimensions for the vital storage of microsurgical free flaps: an experimental approach.

A revolutionary circulatory system has been developed to nourish big, free osteomusculocutaneous flaps extracorporally. Thus we will be able to transplant the free flaps to defect areas that have no sufficient vascular situation. In respect to the cold storage of microsurgical free flaps, to date maximal periods of ischaemic tolerance have been considerably exceeded; the maximal period is currently 168 hours. The vitality of the flap is monitored through parameters setting forth the consumption of oxygen together with histology and electron microscopy. The oxygenation of the nutritive medium is achieved through an aqueous phase gas exchange.

Animals↗

Effect of cold air on exercise capacity in COPD: increase or decrease?

STUDY OBJECTIVE: To clarify the effect of cold air on exercise capacity in COPD. DESIGN: Cycle ergometer tests under different environmental conditions. SETTING: Pulmonary function laboratory and an environmental chamber at a university hospital. PARTICIPANTS: Eighteen patients with stable COPD; 14 completed the study. INTERVENTIONS: A preliminary cycle ergometer test followed by two incremental, symptom-limited cycle ergometer tests, one at 24 degrees C and the other at -20 degrees C. MEASUREMENTS: On the first study day: arterial blood gas analysis, 12 to 15 s maximal voluntary ventilation, maximal expiratory flow-volume curves before and 1 h after inhalation of 80 microg of ipratropium bromide, and diffusion capacity of the lung. During the exercise challenges: spirometric indices, minute ventilation (VE), oxygen consumption (VO2), carbon dioxide production (VCO2), facial skin temperature, and heart rate. The feeling of dyspnea was assessed with a visual analogue scale. RESULTS: The maximal work load was 87.5+/-7.3 W at -20 degrees C compared with 96.4+/-6.9 W at 24 degrees C (p<0.05). Accordingly, the exercise duration was shorter in the cold. Exercise dyspnea was more severe in the cold at equal work loads. The shortening of exercise duration induced by cold air correlated with the enhancement of exercise dyspnea. Furthermore, cold air cooled the facial skin and induced immediate bronchoconstriction. VE, VO2, VCO2, and heart rate did not differ between the warm and cold challenges. CONCLUSIONS: Cold air decreases exercise capacity in COPD, probably by increasing exercise dyspnea.

Adult↗

Role of influenza vaccine for healthy children in the US.

Influenza infection is associated with significant morbidity and mortality in adults, but the highest attack rates for influenza regularly occur in children, particularly those in preschool and elementary school. The consequences of influenza in this younger population - increased rate of hospitalization in those younger than 2 years of age and serious associated morbidity - have been underestimated. Children are also the critical link for spreading influenza in the community. Recent data suggest that mass influenza vaccination of healthy children would not only protect recipients, but also may reduce the burden of influenza throughout the community. During the past 3 decades, efforts to control influenza have focused on the use of an injectable trivalent inactivated vaccine (TIV) in high-risk persons. The vaccine is 'safe' and effective, but its acceptance and uptake by patients and healthcare providers have been modest at best. A new intranasal, live-attenuated, trivalent cold-adapted influenza virus vaccine (CAIV-T) [FluMist] is 'safe', well tolerated, immunogenic, and efficacious in preventing influenza illness in healthy children. Compared with TIV, CAIV-T is easier to administer and should be more readily acceptable, particularly for mass immunization campaigns. CAIV-T also induces a broader immune response and has demonstrated protection against at least three different variant influenza strains. This vaccine is particularly well suited for routine immunization of children and thus offers the potential for greatly improved control of influenza. However, the acquisition cost per single dose of FluMist for the 2003-4 season ( approximate, equals 46 US dollars) significantly hampered its uptake both by practitioners and by managed care organizations, even despite a later approximate, equals 25 US dollars rebate offer. For the 2004-5 season, CAIV-T is likely to be only modestly more expensive (average wholesale price: 16.50 US dollars for non-returnable doses, 23 US dollars for returnable doses) than TIV. The practitioner must consider the benefits of FluMist compared with its likely higher vaccine cost and the issues of reimbursement among multiple insurers.

Adolescent↗

In vitro thymus suppression of hemagglutinin production in Xenopus laevis: location, drug and temperature sensitivity.

Suppression of splenic hemagglutinin (HA) production by allothymuses in vitro was further studied in Xenopus laevis, the South African clawed toad. Since thymic capacity to suppress splenic HA secretion into the culture medium is retained in animals previously exposed to N-methyl-N-nitrosourea (NMU), which destroys the thymus cortex in this species, suppressor function must be located in the thymus medulla. Reciprocal thymus-spleen combinations showed that normal thymus can suppress immunised spleen fragments from NMU-treated animals. Since Xenopus exposed to NMU are also devoid of helper function, thymus suppression acts directly on the antibody forming system. Cyclophosphamide removed thymic suppression and enhanced spleen fragment antibody production, while Cyclosporin A enhanced thymic suppression and blocked HA production. Therefore, antigenic stimulation of thymic suppression and antibody production are inversely related in this species. Long-term storage of Xenopus in the cold increased lymphoid cellularity of the thymus medulla, but thymuses from cold-stored animals could not suppress normal spleen fragments and their spleens were not suppressed by normal thymuses. Thus, ectotherms may retain immunological capacity when subjected to prolonged cold by a loss of thymic suppression.

Animals↗

[Physiological bases for work and rest schedules during shift work in Arctic tundra].

Conditions of Far North determine use of shifted work mode for construction of railways. The studies proved that low-distance shifted work is the most acceptable due to preserved health and work performance, lower rate of ageing. This mode maintains adaptation to the region's conditions, high level of work performance, saves the qualified workers.

Adaptation, Physiological↗

Review: clinical opportunities provided by the nasal administration of peptides.

Peptides are rapidly being developed as potential new therapeutic agents and the nasal route is being evaluated as a means of achieving systemic absorption. Current research in man is being directed at a number of polypeptides, including calcitonin, growth hormone releasing hormones (GHRH), insulin, gonadotropin hormone releasing hormones (GnRH) and vasopressin analogues. The underlying protective functions of the nose provide anatomical, temporal and enzymatic barriers to absorption of peptides. The nasal route is relatively unsuccessful when used for high molecular weight polypeptides. Penetration enhancers improve bioavailability but are poorly tolerated. Reproducibility of effect is highly variable, major contributing factors including the site of deposition and type of delivery system as well as changes in the mucous secretion and mucociliary clearance, compounded by the presence of allergy, hay fever and the common cold in treated subjects. The future potential for this route lies in development of effective and well tolerated formulations in highly accurate delivery systems for the chronic administration of peptides, enabling the replacement of impractical and invasive intravenous injections in patients on lifelong substitution treatment for various deficiency states.

Administration, Intranasal↗

Post exercise changes in compartmental body temperature accompanying intermittent cold water cooling in the hyperthermic horse.

Whereas the efficacy of cold water cooling of horses has been demonstrated by several studies, the dynamics of temperature changes within and between compartments (primarily muscle, blood [core], skin and deep core [rectal]) have not been investigated. Changes in body temperature associated with cold water cooling were investigated in the hyperthermic horse. Muscle (TMU), pulmonary artery (TPA), rectal (TREC), tail-skin (TTSK) and coat surface (TCOAT) temperatures, were monitored continuously in 5 Thoroughbred horses during and after exercise in hot humid (30 degrees C and 80% RH) conditions on a treadmill. Horses were cooled in the hot humid environment with cold water (approximately 6 degrees C) for 6 30 s periods. Between each 30 s cooling period the horses stood for 30 s. A total of 180 l of cold water was applied. Horses were monitored for a further 4 min following the final cooling period. From the end of exercise to the end of the final cooling (6.5 min), mean (+/- s.e.) rates of decrease for TTSK and TPA were similar (0.8 +/- 0.1 and 0.8 +/- 0.1 degrees C/min, respectively). The effects on TMU and TREC were less marked, with average rates of 0.2 +/- 0.1 and 0.0 +/- 0.1 degrees C/min, respectively. During the first 4 min of cooling, TPA fell during the 30 s period of water application and rose during each 30 s period of standing. When TPA fell below approximately 36.5 degrees C, these variations were suppressed and TPA rose steadily, despite continued applications; TREC and TMU continued to fall, although less rapidly than before. These observations are consistent with the onset of skin vasoconstriction at low TPA. The mechanism is mediated through a cooling of circulating blood volume providing a greater capacity for heat transfer between muscle and circulation. Intermittent application of cold water (approximately 6 degrees C) improves heat removal without apparent deleterious effects and is well tolerated. Even when hypothermia develops (based on TPA), muscle and rectal temperatures continue to fall.

Animals↗

Effects of caffeine and various xanthines on hornets and bees.

The effect of caffeine was assessed on Vespa orientalis hornets maintained either in sealed breeding boxes or as entire colonies free to forage, and also on Apis mellifera bees within their hives. In a number of instances the hornets were also used to study the effect of various bodily extracts of queen hornets and of the following xanthines: Purine; hypoxanthine; uric acid; theophylline; and theobromine. The studied materials were found to exert an effect on three categories of activities: (1) Motor motility, flight, and construction; (2) sensory response to light (retinal and extraretinal), noise, irritability, orientation; and (3) physiological changes in appetite, copulation, oviposition, hibernation, resistance to cold, and longevity. Up to a point the produced effects were reversible. Throughout the period of experimentation the test insects did not show signs of tolerance or addiction towards caffeine.

Animals↗

Overexpression of multiple dehydrin genes enhances tolerance to freezing stress in Arabidopsis.

To elucidate the contribution of dehydrins (DHNs) to freezing stress tolerance in Arabidopsis, transgenic plants overexpressing multiple DHN genes were generated. Chimeric double constructs for expression of RAB18 and COR47 (pTP9) or LTI29 and LTI30 (pTP10) were made by fusing the coding sequences of the respective DHN genes to the cauliflower mosaic virus 35S promoter. Overexpression of the chimeric genes in Arabidopsis resulted in accumulation of the corresponding dehydrins to levels similar or higher than in cold-acclimated wild-type plants. Transgenic plants exhibited lower LT50 values and improved survival when exposed to freezing stress compared to the control plants. Post-embedding immuno electron microscopy of high-pressure frozen, freeze-substituted samples revealed partial intracellular translocation from cytosol to the vicinity of the membranes of the acidic dehydrin LTI29 during cold acclimation in transgenic plants. This study provides evidence that dehydrins contribute to freezing stress tolerance in plants and suggests that this could be partly due to their protective effect on membranes.

Adaptation, Physiological↗

Comparison of the cold hardiness capacities of the oviparous and viviparous forms of Lacerta vivipara.

The lizard Lacerta vivipara has allopatric oviparous and viviparous populations. The cold hardiness strategy of L. vivipara has previously been studied in viviparous populations, but never in oviparous ones. The present study reveals that both the oviparous and viviparous individuals of this species are able to survive in a supercooled state at -3 degrees C for at least one week when kept on dry substrates. The mean crystallisation temperatures of the body, around -4 degrees C on dry substrata and -2 degrees C on wet substrata, do not differ between oviparous and viviparous individuals. All the individuals are able to tolerate up to 48-50% of their body fluid converted into ice, but only viviparous individuals were able to stabilize their body ice content at 48%, and hence were able to survive even when frozen at -3 degrees C for times of up 24 hours. Ice contents higher than 51% have been constantly found lethal for oviparous individuals. This suggests that, in L. vivipara, the evolution towards a higher degree of freezing tolerance could parallel the evolution of the viviparous reproductive mode, a feature believed to be strongly selected under cold climatic conditions. This is the first report, among reptiles, of an intraspecific variation regarding the freeze tolerance capacities.

Adaptation, Physiological↗

Prazosin treatment of primary Raynaud's phenomenon.

In a double blind study, prazosin, a specific adrenergic alpha 1-receptor antagonist, or placebo were given to 15 females with primary Raynaud's phenomenon. At a low dose (1 mg twice daily) 5 out of 7 of the prazosin-treated patients reported a reduction of attacks induced by cold (p less than 0.05). This was not confirmed by a cold provocation test which showed no improvement at finger temperatures of 15 or 10 degrees C. The highest tolerated dose in the prazosin-treated patients varied from 2-8 mg daily, and the greatest number of side effects was recorded in this group (p less than 0.05). None of the patients experienced complete relief from cold-induced attacks. It was concluded either that Raynaud's phenomenon is not only caused by stimulation of alpha 1-receptors in digital arteries or the clinically achievable blockade was insufficient to prevent attacks.

Blood Pressure↗