Aging preferentially affects molecular pathways implicated in development and disease of myelinating glial cells

The central and peripheral nervous systems are involved in multiple age-dependent neurological deficits that are often attributed to alterations in function of myelinating glial cells. However, the molecular events that underlie the age-related decline of glial cell function are unknown. We used Schwann cells as a model to study biological processes affected in glial cells by aging. We comprehensively profiled gene expression of the Schwann cellrich mouse sciatic nerve throughout life, from day of birth until senescence (840 days of age). We combined the aging data with the microarray transcriptional data obtained using nerves isolated from Schwann cell-specific neuropathy-inducing mutants MPZCre/+/Lpin1fE2−3/fE2−3 , MPZCre/+/ScapfE1/fE1 and Pmp22-null mice. The majority of age related transcripts were also affected in the analyzed mouse models of neuropathy (54.4%) and in development (59.5%) indicating a high level of overlapping in implicated molecular pathways. We observed that compared to peripheral nerve development, dynamically changing expression profiles in aging have opposite (anticorrelated) orientation while they copy the orientation of transcriptional changes observed in analyzed neuropathy models. Subsequent clustering and biological annotation of dynamically changing transcripts revealed that the processes most significantly deregulated in aging include inflammatory/immune response and lipid biosynthesis/metabolism. Importantly, the changes in these pathways were also observed in myelinated oligodendrocyte-rich optic nerves of aged mice, albeit with lower magnitude. This observation suggests that similar biological processes are affected in aging glial cells in central and peripheral nervous systems, however with different dynamics. Our data, which provide the first comprehensive comparison of molecular changes in glial cells in three distinct biological conditions comprising development, aging and disease, provide not only a new inside into the molecular alterations underlying neural system aging but also identify target pathways for potential therapeutic approaches to prevent or delay complications associated with age-related and inherited forms of neuropathies. *Current address: Department of Physiology, UCSF, San Francisco, CA, USA.

Decomposition of Ruppia cirrhosa (Petagna) Grande in the sediment of a coastal lagoon

The decomposition process of Ruppia cirrhosa was studied in a Mediterranean coastal lagoon in the Delta of the River Ebro (NE Spain). Leaves and shoots of Ruppia were enclosed in 1 mm-mesh and 100 pm-mesh litter bags to ascertain the effect of detritivores, macroinvertebrates, and bacteria and fungi, respectively. Changes in biomass and carbon, and, nitrogen and phosphorus concentrations in the detritus were studied at the sediment-water interface and in the sediment. Significant differences in biomass decay were observed between the two bag types. Significant differences in decomposition were observed between the two experimental conditions studied using 100 pm-mesh bags. These differences were not significant when using the 1 mm-mesh bags. The carbon content in the detritus remained constant during the decomposition process. The percentage of nitrogen increased progressively from an initial 2.4 % to 3 %. The percentage of phosphorus decreased rapidly during the first two days of decomposition from an initial 0.26 % to 0.17 %. This loss is greater in the sediment than in the water column or at the sediment-water interface. From these results we deduce that the activity of microorganisms seems to be more important in the sediment than in the water-sediment interface, and that grazing by macroinvertebrates has less importance in the sediment than in the water column.

Effect of nitrogen and phosphorus supply on growth, chlorophyll content and tissue composition of the macroalga Chaetomorpha linum (O.F. Mull), Kutz, in a Mediterranean Coastal Lagoon

The effect of dissolved nutrients on growth, nutrient content and uptake rates of Chaetomorpha linum in a Mediterranean coastal lagoon (Tancada, Ebro delta, NE Spain) was studied in laboratory experiments. Water was enriched with distinct forms of nitrogen, such as nitrate or ammonium and phosphorus. Enrichment with N, P or with both nutrients resulted in a significant increase in the tissue content of these nutrients. N-enrichment was followed by an increase in chlorophyll content after 4 days of treatment, although the difference was only significant when nitrate was added without P. P-enrichment had no significant effect on chlorophyll content. In all the treatments an increase in biomass was obseved after 10 days. This increase was higher in the N+P treatments. In all the treatments the uptake rate was significantly higher when nutrients were added than in control jars. The uptake rate of N, as ammonium, and P were significantly higher when they were added alone while that of N as nitrate was higher in the N+P treatment. In the P-enriched cultures, the final P-content of macroalgal tissues was ten-fold that of the initial tissue concentrations, thereby indicating luxury P-uptake. Moreover, at the end of the incubation the N:P ratio increased to 80, showing that P rather than N was the limiting factor for C. linum in the Tancada lagoon. The relatively high availability of N is related to the N inputs from rice fields that surround the lagoon and to P binding in sediments.

Evaluation of the integrity of human corneal epithelium maintained in organ-culture during eye banking using Corneamax®

Purpose: Mediums have been developed to conserve corneal endothelium in organ-culture during eye banking. CorneaMax® is used by 25% of Eye Bank in Europe. Only little is known about conservation of corneal epithelium with this medium during banking. Its preservation could be of interest in clinic to cure corneal disease with stem cells deficiency. Therefore, we wanted to examine the integrity of human corneal epithelium maintained in CorneaMax®. Methods: Human corneas, considered unsuitable for transplantation, were obtained from the Eye Bank in Lausanne. Average post-mortem time was 14 hours. Cornoscleral rings were maintained in organ-culture in Corneamax® at 32°C. Samples were formalin-fixed after period ranging from 0 (D0) to 35 days (D35, N=5 for each time points) and stained with H&E. Proliferation and apoptosis were evaluated by immunostaining with antibody against Ki67 and Caspase3 respectively. Results: Corneas, which were not in organ-cultured (D0), showed different morphology, including intact epithelium with 5 to 7 layers, but also completely denuded basement membrane. In two cases, at D0, the epithelium lost its adherence to the basal lamina of the cornea creating a large epithelial sheet. During the two first days, corneas and limbus area lost totally their epithelium, except for some remaining limbal basal cells. From day 2 to day 10, regeneration of the epithelium took place, starting from the limbal region in direction to the central cornea. From day 10 to day 35, corneal epithelium appeared as an atrophic epithelium, consisting of only two cell layers. Proliferation happened in the whole cornea during the 35 days of organ-culture, as shown by Ki67 positive cells. Apoptosis was rarely detected in the corneal epithelium. Conclusions: Corneas maintained in CorneaMax® showed a complete disappearance of the corneal epithelium during the two first days and a conservation of limbal basal cells in the limbal region. These remaining cells allowed a full regeneration of the tissue, leading to an atrophic epithelium, composed of only two cell layers. This atrophic epithelium could be seen in all the organ-cultured corneas during the 35 days of conservation. This study is a first step to develop medium in organ-culture in order to conserve corneal epithelial cells.

The direct effects of tacrolimus and cyclosporin A on isolated human islets: A functional, survival and gene expression study.

The use of immunosuppressive drugs in transplanted patients is associated with the development of diabetes, possibly due to β-cell toxicity. To better understand the mechanisms leading to post-transplant diabetes, we investigated the actions of prolonged exposure of isolated human islets to therapeutical levels of tacrolimus (Tac) or cyclosporin A (CsA). Islets were isolated from the pancreas of multiorgan donors by enzymatic digestion and density gradient centrifugation. Functional, survival and molecular studies were then performed after 4 days of incubation with therapeutical concentrations of Tac or  CsA. Glucose-induced insulin secretion was significantly decreased in Tac, but not in CsA exposed islets, which was associated with a reduction of the amount of insulin granules as shown by electron microscopy. The percentage of apoptotic β-cells was higher in Tac than CsA exposed islets. Microarray experiments followed by Gene Set Enrichment Analysis revealed that gene expression was more markedly affected upon Tac treatment. In conclusion, Tac and CsA affect features of beta-cell differently, with several changes occurring at the molecular level.

Effects of packaging and temperature on postharvest of atemoya

Effects of refrigerated storage and different packages on postharvest behaviour of fruits of atemoya (Annona cherimola x A. squamosa) cv. PR3 were investigated. Fruits were individually sealed in copolymer (PD-955) and low-density polyethylene (LDPE) bags and stored for 21 days at 15°C or 25°C. Then they were unwrapped and maintained at 25°C, for ripening. Weight loss and firmness of fruits were both affected by storage time, temperature and packaging. Weight loss in packaged atemoyas was lower than in the control, non-wrapped fruits. A non-trained panel scored the atemoyas for overall eating quality and appearance. Fruits sealed in LDPE did not ripen, probably due to an injurious atmosphere developed inside the package. Atemoyas packaged in PD-955 film had a shelf-life of 17 days against 13 days of the control ones, both stored at 15°C, an increase of 30% on shelf-life.

Effect of growth selection on morphology in Pirenaica cattle

Twelve variables were identified to define morphology in 109 calves sired by eight Pirenaica bulls widely used in artificial insemination. The effect of selection for weight at 210 days of age (W210) was detected from the regression coefficient between the 12 variables and the selection index for W210 used in the breeding plan. Unbiased estimates of the genetic correlated responses on the morphology were obtained without previously estimating the genetic correlation among traits. In Pirenaica cattle, selection for W210 increased slaughter live weight, chest depth and corporal length. The expected changes on height (withers, back and rump), rump width and thoracic perimeter were lower but positive and statistically significant.

Aging preferentially affects molecular pathways implicated in development and disease of myelinating glial cells

The central and peripheral nervous systems are involved in multiple agedependent neurological deficits that are often attributed to alterations in function of myelinating glial cells. However, the molecular events that underlie the age-related decline of glial cell function are unknown. We used Schwann cells as a model to study biological processes affected in glial cells by aging. We comprehensively profiled gene expression of the Schwann cell-rich mouse sciatic nerve throughout life, from day of birth until senescence (840 days of age). We combined the aging data with the microarray transcriptional data obtained using nerves isolated from Schwann cell-specific neuropathy-inducing mutants MPZCre/þ/Lpin1fE2-3/fE2-3, MPZCre/þ/ScapfE1/fE1 and Pmp22-null mice. A majority of age related transcripts were also affected in the analyzed mouse models of neuropathy (54.4%) and in development (59.5%) indicating a high level of overlapping in implicated molecular pathways. We observed that compared to peripheral nerve development, dynamically changing expression profiles in aging have opposite (anticorrelated) orientation while they copy the orientation of transcriptional changes observed in analyzed neuropathy models. Subsequent clustering and biological annotation of dynamically changing transcripts revealed that the processes most significantly deregulated in aging include inflammatory/ immune response and lipid biosynthesis/metabolism. Importantly, the changes in these pathways were also observed in myelinated oligodendrocyte- rich optic nerves of aged mice, albeit with lower magnitude. This observation suggests that similar biological processes are affected in aging glial cells in central and peripheral nervous systems, however with different dynamics. Our data, which provide the first comprehensive comparison of molecular changes in glial cells in three distinct biological conditions comprising development, aging and disease, provide not only a new inside into the molecular alterations underlying neural system aging but also identify target pathways for potential therapeutical approaches to prevent or delay complications associated with age-related and inherited forms of neuropathies.

Induction of somatic embryogenesis in immature seeds of guavatree cv. Paluma

The biotechnological techniques may help solve many problems of guava culture, such as the high perishability of fruits. Somatic embryogenesis can generate highly multiplicative cell cultures and with high regenerative potential, serving as basis for genetic transformation. The aim of this work was to obtain somatic embryogenesis of guava (Psidium guajava L.) cv. Paluma. Immature seeds were used, and they were inoculated in MS environment containing 400 mg L-1 of L-glutamine, 100 mg L-1 myo-inositol, 60 g L-1 sucrose, 100 mg L-1 ascorbic acid and supplemented with different types and concentrations of growth regulators. Embryogenic callus appeared after 37 days of culture in an environment containing 1.0 mg L-1 2.4-D + 2.0 mg L-1 2-ip, in 7% of the explants. After 65 days of culture, the treatment containing 0.5 mg L-1 CPA showed 20% of explants with direct embryos, while the treatment with 1 mg L-1 had 14% of explants with direct embryos and 7% of explants with embryogenic callus. In 66.6% of embryos regenerated with 0.5 mg L¹ CPA there was the formation of secondary embryos. The use of IASP and BAP, aiming embryogenesis proliferation, led to an increase in the cellular proliferation, but calli apparently lost their embryogenic potential.

Residual effect of growth regulators in etiolation and regeneration of in vitro pineapple plants

This work aimed to evaluate the influence of naphthaleneacetic acid (NAA) and gibberellic acid (GA3) plant regulators in in vitro etiolation and subsequent regeneration of the PE x SC-60 pineapple hybrid. Nodal segments of in vitro plants with approximately 5-7 cm height were incubated in basic MS culture medium supplemented with 0.0; 0.5 and 1.0 mg L-1 of naphthaleneacetic acid (NAA) in combination with gibberellic acid (GA3) in concentrations of 0.0; 0.5 and 1.0 mg L-1, and maintained at 27 ºC under dark condition. Evaluations were carried out at 90 and 180 days after incubation period. The best results for length of etiolated stems were obtained with 1.0 mg L-1 of NAA. In the experiment followed by the regeneration, stems with 3 cm from the etiolation treatment, were cultivated in proliferation medium and the number of regenerated plants per treatment was evaluated at 60 days of cultivation. The treatment that promoted the best etiolation of plants also promoted the worst regeneration rates, demonstrating the residual effect of the auxin used in the previous step in the regeneration of plants of the pineapple hybrid evaluated.

Sensory characteristics of 'Douradão' peaches submitted to modified atmosphere packaging

The sensory, physical and chemical characteristics of 'Douradão' peaches cold stored in different modified atmosphere packaging (LDPE bags of 30, 50, 60, 75µm thickness) were studied. After 14, 21 and 28 days of cold storage (1 ± 1 ºC and 90 ± 5% RH), samples were withdrawn from MAP and kept during 4 days in ambient air for ripening. Descriptive terminology and sensory profile of the peaches were developed by methodology based on the Quantitative Descriptive Analysis (QDA). The assessors consensually defined the sensory descriptors, their respective reference materials and the descriptive evaluation ballot. Fourteen individuals were selected as judges based on their discrimination capacity and reproducibility. Seven descriptors were generated showing similarities and differences among the samples. The data were analysed by ANOVA, Tukey test and Principal Component Analysis (PCA). The atmospheres that developed inside the different packaging materials during cold storage differed significantly. The PCA showed that MA50 and MA60 treatments were more characterized by the fresh peach flavour, fresh appearance, juiciness and flesh firmness, and were effective for keeping good quality of 'Douradão' peaches during 28 d of cold storage. The Control and MA30 treatments were characterized by the mealiness, the MA75 treatment showed lower intensity for all attributes evaluated and they were ineffective to maintain good quality of the fruits during cold storage. Higher correlation coefficients (positive) were found between fresh appearance and flesh firmness (0.95), fresh appearance and juiciness (0.97), ratio and intensity of fresh peach smell (0.81), as well as higher correlation coefficients (negative) between Hue angle and intensity of yellow colour (-0.91), fresh appearance and mealiness (-0.92), juiciness and mealiness (-0.95), firmness and mealiness (-0.94).

Modified atmosphere packaging extending the storage life of 'douradão' peach

'Douradão' peach is a perishable product and when cold stored is subject to chilling injury. The objective of the experiment was to evaluate the effect of modified atmosphere packaging (MAP) and cold storage on quality and storage life of these peaches. Fruits were packed in polypropylene (PP) trays and placed inside low density polyethylene (LDPE) bags (30, 50, 60, 75 μm thickness) with active modified atmosphere (10 kPa CO2 + 1.5kPa O2, balance N2). The control was made with peaches held in nonwrapped PP trays. Fruits were kept at 1 ± 1 °C and 90 ± 5% relative humidity (RH) for 28 days and CO2 and O2 within packages was monitored every two days. After 14, 21 and 28 days, samples were withdrawn from MAP and kept in air at 25 ± 1 °C and 90 ± 5% RH for ripening. On the day of removal from the cold storage and after 4 days, peaches were evaluated for weight loss, decay incidence, flesh firmness, woolliness incidence, soluble solids content (SSC), titratable acidity (TA) and juice content. The results showed that MAP had influence on reducing weight loss and prevented postharvest decay. MAP of 1-2 kPa O2 and 3-6 kPa CO2 at 1 °C (from 50 and 60 μm LDPE films) were effective for keeping good quality of 'Douradão' peaches during 28 days of storage, the ripe fruits showed reduced incidence of woolliness, adequate juiciness and flesh firmness. Packages of 30 and 75 μm LDPE films were ineffective for reducing woolliness during cold storage. MAP fruits showed lower SSC and no relevant effect on TA. Control fruits did not present marketable conditions after 14 days of cold storage.

Effects of modified atmosphere packaging on ripening of 'Douradão' peach related to pectolytic enzymes activities and chilling injury symptoms

The present study evaluated the effects of modified atmosphere packaging on inhibition of the development of chilling injury symptoms in 'Douradão' peach after cold storage and the possible involvement of cell wall enzymes. Fruits were harvested at the middle stadium of ripening, packed in polypropylene trays and placed inside low density polyethylene (LDPE) bags (30, 50, 60 and 75 µm of thickness) with active modified atmosphere (10 kPa CO2 + 1.5 kPa O2, balance N2). The following treatments were tested: Control: peaches held in nonwrapped trays; MA30: LDPE film - 30 µm; MA50: LDPE film - 50 µm; MA60: LDPE film - 60 µm and MA75: LDPE film - 75 µm. Fruits were kept at 1±1ºC and 90±5% relative humidity (RH) for 28 days. After 14, 21 and 28 days, samples were withdrawn from MAP and kept in air at 25±1ºC and 90±5% RH for ripening. On the day of removal and after 4 days, peaches were evaluated for woolliness incidence, pectolytic enzymes activities. The respiratory rate and ethylene synthesis were monitored during 6 days of ripening. The results showed that MA50 and MA60 treatments had positive effect on the inhibition of the development of woolly texture and reduced pectin methylesterase activity on the ripe fruits, keeping good quality of 'Douradão' peach during 28 days of cold storage. The treatments Control, MA30 and MA75 showed higher woolliness incidence and did not present marketable conditions after 14 days of cold storage.

The effect of dose on the antimalarial efficacy of artemether-lumefantrine: a systematic review and pooled analysis of individual patient data.

BACKGROUND: Artemether-lumefantrine is the most widely used artemisinin-based combination therapy for malaria, although treatment failures occur in some regions. We investigated the effect of dosing strategy on efficacy in a pooled analysis from trials done in a wide range of malaria-endemic settings. METHODS: We searched PubMed for clinical trials that enrolled and treated patients with artemether-lumefantrine and were published from 1960 to December, 2012. We merged individual patient data from these trials by use of standardised methods. The primary endpoint was the PCR-adjusted risk of Plasmodium falciparum recrudescence by day 28. Secondary endpoints consisted of the PCR-adjusted risk of P falciparum recurrence by day 42, PCR-unadjusted risk of P falciparum recurrence by day 42, early parasite clearance, and gametocyte carriage. Risk factors for PCR-adjusted recrudescence were identified using Cox's regression model with frailty shared across the study sites. FINDINGS: We included 61 studies done between January, 1998, and December, 2012, and included 14 327 patients in our analyses. The PCR-adjusted therapeutic efficacy was 97·6% (95% CI 97·4-97·9) at day 28 and 96·0% (95·6-96·5) at day 42. After controlling for age and parasitaemia, patients prescribed a higher dose of artemether had a lower risk of having parasitaemia on day 1 (adjusted odds ratio [OR] 0·92, 95% CI 0·86-0·99 for every 1 mg/kg increase in daily artemether dose; p=0·024), but not on day 2 (p=0·69) or day 3 (0·087). In Asia, children weighing 10-15 kg who received a total lumefantrine dose less than 60 mg/kg had the lowest PCR-adjusted efficacy (91·7%, 95% CI 86·5-96·9). In Africa, the risk of treatment failure was greatest in malnourished children aged 1-3 years (PCR-adjusted efficacy 94·3%, 95% CI 92·3-96·3). A higher artemether dose was associated with a lower gametocyte presence within 14 days of treatment (adjusted OR 0·92, 95% CI 0·85-0·99; p=0·037 for every 1 mg/kg increase in total artemether dose). INTERPRETATION: The recommended dose of artemether-lumefantrine provides reliable efficacy in most patients with uncomplicated malaria. However, therapeutic efficacy was lowest in young children from Asia and young underweight children from Africa; a higher dose regimen should be assessed in these groups. FUNDING: Bill & Melinda Gates Foundation.

AltitudeOmics : impaired pulmonary gas exchange efficiency and blunted ventilatory acclimatization in humans with patent foramen ovale after 16 days at 5,260 m.

A patent foramen ovale (PFO), present in ∼40% of the general population, is a potential source of right-to-left shunt that can impair pulmonary gas exchange efficiency [i.e., increase the alveolar-to-arterial Po2 difference (A-aDO2)]. Prior studies investigating human acclimatization to high-altitude with A-aDO2 as a key parameter have not investigated differences between subjects with (PFO+) or without a PFO (PFO-). We hypothesized that in PFO+ subjects A-aDO2 would not improve (i.e., decrease) after acclimatization to high altitude compared with PFO- subjects. Twenty-one (11 PFO+) healthy sea-level residents were studied at rest and during cycle ergometer exercise at the highest iso-workload achieved at sea level (SL), after acute transport to 5,260 m (ALT1), and again at 5,260 m after 16 days of high-altitude acclimatization (ALT16). In contrast to PFO- subjects, PFO+ subjects had 1) no improvement in A-aDO2 at rest and during exercise at ALT16 compared with ALT1, 2) no significant increase in resting alveolar ventilation, or alveolar Po2, at ALT16 compared with ALT1, and consequently had 3) an increased arterial Pco2 and decreased arterial Po2 and arterial O2 saturation at rest at ALT16. Furthermore, PFO+ subjects had an increased incidence of acute mountain sickness (AMS) at ALT1 concomitant with significantly lower peripheral O2 saturation (SpO2). These data suggest that PFO+ subjects have increased susceptibility to AMS when not taking prophylactic treatments, that right-to-left shunt through a PFO impairs pulmonary gas exchange efficiency even after acclimatization to high altitude, and that PFO+ subjects have blunted ventilatory acclimatization after 16 days at altitude compared with PFO- subjects.