An extremely young massive clump forming by gravitational collapse in a primordial galaxy.

When cosmic star formation history reaches a peak (at about redshift z ≈ 2), galaxies vigorously fed by cosmic reservoirs are dominated by gas and contain massive star-forming clumps, which are thought to form by violent gravitational instabilities in highly turbulent gas-rich disks. However, a clump formation event has not yet been observed, and it is debated whether clumps can survive energetic feedback from young stars, and afterwards migrate inwards to form galaxy bulges. Here we report the spatially resolved spectroscopy of a bright off-nuclear emission line region in a galaxy at z = 1.987. Although this region dominates star formation in the galaxy disk, its stellar continuum remains undetected in deep imaging, revealing an extremely young (less than ten million years old) massive clump, forming through the gravitational collapse of more than one billion solar masses of gas. Gas consumption in this young clump is more than tenfold faster than in the host galaxy, displaying high star-formation efficiency during this phase, in agreement with our hydrodynamic simulations. The frequency of older clumps with similar masses, coupled with our initial estimate of their formation rate (about 2.5 per billion years), supports long lifetimes (about 500 million years), favouring models in which clumps survive feedback and grow the bulges of present-day galaxies.

Hospital surge capacity in a tertiary emergency referral centre during the COVID-19 outbreak in Italy.

The first person-to-person transmission of the 2019 novel coronavirus in Italy on 21 February 2020 led to an infection chain that represents one of the largest known COVID-19 outbreaks outside Asia. In northern Italy in particular, we rapidly experienced a critical care crisis due to a shortage of intensive care beds, as we expected according to data reported in China. Based on our experience of managing this surge, we produced this review to support other healthcare services in preparedness and training of hospitals during the current coronavirus outbreak. We had a dedicated task force that identified a response plan, which included: (1) establishment of dedicated, cohorted intensive care units for COVID-19-positive patients; (2) design of appropriate procedures for pre-triage, diagnosis and isolation of suspected and confirmed cases; and (3) training of all staff to work in the dedicated intensive care unit, in personal protective equipment usage and patient management. Hospital multidisciplinary and departmental collaboration was needed to work on all principles of surge capacity, including: space definition; supplies provision; staff recruitment; and ad hoc training. Dedicated protocols were applied where full isolation of spaces, staff and patients was implemented. Opening the unit and the whole hospital emergency process required the multidisciplinary, multi-level involvement of healthcare providers and hospital managers all working towards a common goal: patient care and hospital safety. Hospitals should be prepared to face severe disruptions to their routine and it is very likely that protocols and procedures might require re-discussion and updating on a daily basis.

Recruitment manoeuvres dislodge mucus towards the distal airways in an experimental model of severe pneumonia.

BACKGROUND: Recruitment manoeuvres generate a transient increase in trans-pulmonary pressure that could open collapsed alveoli. Recruitment manoeuvres might generate very high inspiratory airflows. We evaluated whether recruitment manoeuvres could displace respiratory secretions towards the distal airways and impair gas exchange in a porcine model of bacterial pneumonia. METHODS: We conducted a prospective randomised study in 10 mechanically ventilated pigs. Pneumonia was produced by direct intra-bronchial introduction of Pseudomonas aeruginosa. Four recruitment manoeuvres were applied randomly: extended sigh (ES), maximal recruitment strategy (MRS), sudden increase in driving pressure and PEEP (SI-PEEP), and sustained inflation (SI). Mucus transport was assessed by fluoroscopic tracking of radiopaque disks before and during each recruitment manoeuvre. The effects of each RM on gas exchange were assessed 15 min after the intervention. RESULTS: Before recruitment manoeuvres, mucus always cleared towards the glottis. Conversely, mucus was displaced towards the distal airways in 28.6% ES applications and 50% of all other recruitment manoeuvres (P=0.053). Median mucus velocity was 1.26 mm min-1 [0.48-3.89] before each recruitment manoeuvre, but was reversed (P=0.007) during ES [0.10 mm min-1 [-0.04-1.00]], MRS [0.10 mm min-1 [-0.4-0.48]], SI-PEEP [0.02 mm min-1 [-0.14-0.34]], and SI [0.10 mm min-1 [-0.63-0.75]]. When PaO2 failed to improve after recruitment manoeuvre, mucus was displaced towards the distal airways in 68.7% of the cases, compared with 31.2% recruitment manoeuvres associated with improved PaO2 (odds ratio: 4.76 (95% confidence interval: 1.13-19.97). CONCLUSIONS: Recruitment manoeuvres dislodge mucus distally, irrespective of airflow generated by different recruitment manoeuvres. Further investigation in humans is warranted to corroborate these pre clinical findings, as there may be limited benefits associated with lung recruitment in pneumonia.

Stress during first gestation of ewes impairs memory and learning of male offspring.

This study aimed to investigate the influence of gestational stress induced by lipopolysaccharide (LPS, Escherichia coli) on the physiological changes of ewes, as well as on the subsequent behavioral interaction between ewes and lambs and on the memory and learning of 30-day-old offspring in a T-maze. Thirty-six nulliparous pregnant crossbred Santa Ines ewes with an initial live weight of 45 ± 6 kg, age of 12 ± 2 months, and body condition score between 3 and 3.5 (on a scale of 1 to 5) were divided into two treatments: LPS treatment (E. coli; 0.8 µg.kg-1) and Control (placebo/saline) administered in late pregnancy (day 120). Blood samples were collected before (0 h at 5:00 h) and 1 h, 2 h, 4 h, 8 h, 12 h, 24 h after the administration of LPS or placebo to determine the cortisol release curve. Rectal temperature was measured at the same time points. After birth, male lambs (N = 19) were used to evaluate the maternal-offspring behavioral interaction, weight, and cognitive ability in a T-maze. Blood cortisol and rectal temperature of ewes increased after LPS administration and returned to baseline levels after 24 h. The activities facilitating and stimulating suckling were higher on LPS group (P < 0.05). Lambs whose mothers were challenged with LPS during late pregnancy showed greater learning and memory disabilities including fear behavior and the inability to make decisions at 30 days of age in the T-maze. In sheep, the immunological stress induced by LPS in late pregnancy promotes an inflammatory response characterized by specific rectal temperature and cortisol release profiles, improving maternal care that can increase offspring survival; however, the exposure of sheep fetuses to maternal inflammation causes cognitive impairment in lambs at 30 days of age, which could not be reduced by the behavioral interaction between the mother and offspring.

Evidence of Air Trapping During Ex Vivo Lung Perfusion: A Swine Experimental Lung Imaging and Mechanics Study.

Ex vivo lung perfusion (EVLP) allows the ventilation and perfusion of lungs to evaluate their viability for transplantation. The aim of this study is to compare the mechanical, morphologic and functional properties of lungs during EVLP with values obtained in vivo to guide a safe mechanical ventilation strategy. Lungs from 5 healthy pigs were studied in vivo and during 4 hours of EVLP. Lung compliance, airway resistance, gas exchange, and hemodynamic parameters were collected at positive end-expiratory pressure (PEEP) of 5 cm H2O. Computed tomography was performed at PEEP 0, PEEP 5, and total lung capacity (TLC). Lung pressure-volume (PV) curves were performed from PEEP 0 to TLC. Lung compliance decreased during EVLP (53 ± 5 mL/cm H2O vs 29 ± 7 mL/cm H2O, P < .05), and the PV curve showed a lower inflection point. Gas content (528 ± 118 mL vs 892 ± 402 mL at PEEP 0) and airway resistance (25 ± 5 vs 44 ± 9 cmH2O/L∗s-1, P < .05) were higher during EVLP. Alveolar dead space (5% ± 2% vs 17% ± 6%, P < .05) and intrapulmonary shunt (9% ± 2% vs 28% ± 13%, P < .05) increased ex vivo compared to in vivo, while the partial pressure of oxygen to inspired oxygen fraction ratio (PO2/FiO2) did not differ (468 ± 52 mm Hg vs 536 ± 14 mm Hg). In conclusion, during EVLP lungs show signs of air trapping and bronchoconstriction, resulting in low compliance and increased alveolar dead space. Intrapulmonary shunt is high despite oxygenation levels acceptable for transplantation.

Risk factors for skin lesions on the necks of Norwegian dairy cows.

Physical restrictions at the feed barrier may cause skin lesions, especially on the necks of cows. Thus, some aspects of the functionality of feed barrier design can be assessed by observing skin lesions on the necks of dairy cattle. In this study, 232 herds, mainly of the breed Norwegian Red (94%), were investigated between September 2006 and June 2007. The herds were categorized into 2 groups. One data set, based on 1,148 cows (115 herds), was from farms using feed barriers with post-and-rail designs (86 herds) and combinations of barriers (29 herds). The second data set, which included 1,168 cows (117 herds), was from farms using only vertical feed barriers such as tombstone, diagonal, and headlock. Skin lesions were observed on the necks of 21% of the 2,335 cows observed. Forty-two percent of cows had neck lesions in herds with post-and-rail barriers, whereas 4% of cows had neck lesions in herds with vertical barriers and 30% of cows had neck lesions in herds with a combination of both horizontal and vertical barriers. Alternating logistic regression was used to identify factors associated with skin lesions on the necks of cows. The estimates were transformed into odds ratios (OR). In herds in which post-and-rail design barriers were used, the following risk factors were associated with skin lesions: manger front lower than 63 cm versus higher (OR=3.6), no feed stalls versus feed stalls (OR=10.9), mixed rations versus no mixed ration (OR=3.8), cows later than 30 DIM versus earlier (OR=1.9), cows with low milk production versus those producing 1,000 kg higher (OR=1.4), feeding frequency of less than 3 times daily versus 3 and more times daily (OR=1.9), using only post-and-rail feed barriers (OR=1.8) versus using combinations, and farmer disagreement with the statement "animals experience physical pain as humans do" (OR=1.9) versus agreement. Diagonal barriers, categorized as vertical barriers, increased the association of skin lesions with increasing manger front heights. Feed barrier design was associated with skin lesions on the necks of cows, particularly for barriers of a post-and-rail design. To minimize the risk of neck lesions, the top rail height of post-and-rail barriers should be above 109 cm, especially for cows taller than 126 cm, and the manger front should not be lower than 63 cm. Feed stalls, feeding frequency of 3 or more times daily, and the feeding of unmixed rations were also identified as preventive factors.

Dairy farmer attitudes and empathy toward animals are associated with animal welfare indicators.

Attitudes and empathy of farmers influence human-animal interaction, thereby affecting their behavior toward animals. The goal was to investigate how measures of attitude and empathy toward animals were associated with animal welfare indicators such as milk yield, mastitis incidence, fertility index, and the prevalence of skin lesions on cows. To assess empathy toward animals, a photo-based pain assessment instrument was developed depicting various conditions that could be associated with some degree of pain in cattle and included questions aimed at assessing attitudes toward animals. Photos of painful conditions are useful in eliciting measurable empathic responses to pain in humans. A total of 221 farmers were sampled via e-mail and 154 responses were obtained. In the first analysis, farmers were categorized into 2 groups according to their agreement or disagreement with the attitude statement "animals experience physical pain as humans do." In the second analysis, farmers were assigned a median pain assessment score obtained from their estimates on the visual analog scale of 21 conditions assumed painful for cattle. In the third analysis, farmers were clustered in 3 groups according to their visual analog scale responses. Three conditions were ranked as the most painful: fracture of tuber coxae, dystocia, and serious mastitis. Farmers with positive attitudes toward animals scored 2 points higher on their empathy score compared with farmers with negative attitudes. Personal experience with each additional condition resulted in a 0.09 higher score. Cluster analysis revealed 3 groups. Farmers in group 3 had the highest median pain assessment score (6.7+/-0.2), indicating a high level of empathy and a positive attitude toward animals. They had the lowest prevalence of skin lesions over the carpus (24+/-6%) and the lowest milk production (6,705+/-202 kg). The complex associations between indicators of empathy and attitudes with relevant welfare outcomes suggest that competence building to safeguard animal welfare could benefit from including both attitudes and empathy in human-animal interactions studies.

Prevalence and risk factors for skin lesions on legs of dairy cattle housed in freestalls in Norway.

Appropriate indoor housing facilities for dairy cattle promote improved animal welfare. Skin alterations are an indicator of dysfunctional housing. The purpose was to determine the relationship between different housing design and skin lesions, hence providing farmers more insight into how to reduce the occurrence of lesions. A cross-sectional study was conducted on 2,335 animals in 232 Norwegian freestall-housed dairy cattle from September 2006 to June 2007. A model was established to investigate risk factors related to the presence of lesions including hair loss, swelling, and wounds on the legs of dairy cattle. Separate models were developed to investigate risk factors related to the presence of knee and hock lesions. Previously described risk factors were included in the models, together with a questionnaire addressing the attitude of the farmer toward the animals. Mean (+/- SD) prevalence for hock lesions was 60.5 +/- 21.2% with a median value of 64%. The prevalence for knee lesions was 35.3 +/- 25.7% with a median of 30%. Cows in herds with a soft freestall base had an odds ratio (OR) for knee and hock lesions of 0.22 (0.06 to 0.73) and 0.62 (0.48 to 0.80), respectively, compared with those in herds with a harder freestall base, such as concrete and compact rubber mats. There was an increased risk of hock lesions when the length in the lying area in a double-row freestall was >250 cm (OR = 2.96; 1.02 to 8.60) compared with 260 cm (OR = 2.11; 1.53 to 2.90) compared with < or =260 cm. The risk for knee lesions increased if a row against a wall was >270 cm (OR = 1.72; 1.09 to 2.72) compared with < or =270 cm. Hock lesions were associated with lame cows (OR = 5.76; 1.14 to 29.18) versus nonlame cows, and with cows in their second or higher parity (OR = 1.27; 1.06 to 1.52) versus cows in their first parity. Knee lesions were associated with farmers' negative attitudes toward animals in pain (OR = 3.28; 1.79 to 6.03) versus those with positive attitudes; cows in the beginning of their lactation (OR = 1.84; 1.24 to 2.74) versus those at the end of their lactation; and tall animals (OR = 1.27; 1.00 to 1.61) versus shorter animals. These results show that freestall design is important with respect to skin lesions as are the characteristics of individual animals and the farmer.

Serum biochemistry and electrophoretic patterns in the Eurasian Buzzard (Buteo buteo): reference values.

In avian medicine, hematologic and biochemical laboratory investigations are still in their infancy, because of the difficulty involved in collecting data. This has led to a lack of reference values and a nonstandardized approach to specimens obtained in critical conditions. The Eurasian Buzzard (Buteo buteo) is one of the most common raptors in Italy, yet little is known about the physiologic blood parameters of this species. Serum biochemistry and electrophoretic investigations were performed in 40 healthy Eurasian Buzzards in different Italian wildlife rescue centers waiting to be released after recovering from trauma injuries. Mean values for biochemistry parameters were pancreatic amylase 626.9 IU/l, uric acid 7.5 mg/dl, aspartate aminotransferase 330.9 IU/l, glucose 375.1 mg/dl, lipase 26.3 IU/ l, total protein 38.4 g/l, total bilirubin 0.04 mg/ dl, lactate dehydrogenase 2,008.4 IU/l, creatinine kinase 1,604.1 IU/l, alanine aminotransferase 40.4 IU/l, alkaline phosphatase 89.8 IU/l, magnesium 2.3 mg/dl, calcium 10.2 mg/dl, phosphorus 2.02 mg/dl, cholesterol 192.2 mg/ dl, triglyceride 116.4 mg/dl, albumin 14.5 g/l, creatinine 0.1 mg/dl. Mean electrophoretic values were prealbumin 1.4 g/l, albumin 14.2 g/l, alpha 1 globulin 5.9 g/l, alpha 2 globulin 4.7 g/l, beta globulin 7.5 g/l, gamma globulin 3.6 g/l, albumin/globulins ratio 0.8 g/l.

Attitudes of veterinary students to pain in cattle.

Three hundred veterinary students were divided randomly into two groups to answer a questionnaire inquiring how painful they considered 23 conditions affecting cattle (seven of which were illustrated) to be, on either a numerical rating scale (NRS) or a visual analogue scale (VAS). The individual responses were used to assess whether cluster analysis could be used to divide the population into distinct groups. The ratios of men to women in the two groups were similar. The NRS scores ranged from 1 to 10 and the VAS scores ranged from 0 to 10. The conditions that were illustrated with a picture had a wider range of scores (2.65 to 9.0 v 2.67 to 7.93) and a slightly higher mean score (5.71 v 5.34) than those without pictures. Fracture of the tuber coxae, dystocia requiring the help of two people, and serious mastitis, were scored as the most painful conditions in adult cattle. Using the median score of each student as an outcome, their sex and year of enrollment and the scoring scale were significant. Women scored 0.9 points higher than men. Cluster analysis revealed two distinct groups in both the VAS and NRS, but the distribution was more even among the students using the VAS. This group was used in further analysis, and one cluster had the most men and more students with several siblings.

[The advent of biosimilars: new rules to guarantee patient safety]. / L'avvento dei biosimili: nuove regole per garantire la sicurezza del paziente.

Biosimilars are medicinal products proposed as copies of biotech drugs whose patents have expired. Their entry into the therapeutic armamentarium entails knowledge of the rules established in Europe relevant to their use in clinical practice. In September 2007, an Italian panel of experts comprising two nephrologists, a clinical immunologist, an oncohematologist, a pharmacologist, and a hospital pharmacist examined the main features of biotech drugs and the issues faced by the regulatory authorities in the definition of a specific approval pathway for biosimilars in Europe. The panel of experts agreed that it is important to inform the medical and scientific community that biosimilars are not exact copies of their reference products; therefore the rules governing their clinical use are not the same as those established for biotech drugs in general. Patient safety should be the fundamental principle guiding therapeutic choices, and making these choices should be the prerogative of physicians.

Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines.

Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1ß), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α). This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types. This technique advances traditional ELISAs by: •FMIA provides insight into the profiles of multiple porcine cytokines in certain situations (e.g. disease, parturition).•Use of the Bio-Plex® 200 system to investigate novel sample types, including brain, placenta and synovial tissue.•Multiplexing utilises a fraction of the sample volume compared with multiple ELISAs.

Preliminary Experience With Hypothermic Oxygenated Machine Perfusion in an Italian Liver Transplant Center.

BACKGROUND: Machine perfusion is increasingly utilized in liver transplantation to face the detrimental consequences of the use of extended-criteria donors. Hypothermic oxygenated machine perfusion (HOPE) appears to be more protective relative to static cold storage. Conversely, normothermic machine perfusion (NMP) allows a better graft evaluation. We describe a pilot prospective study on machine perfusion in selected grafts. METHODS: HOPE was executed for all the grafts procured from donors after cardiac death (DCDs) and for livers from donors after brain death (DBDs) requiring prolonged preservation time. NMP was used when a more precise evaluation was needed. Both HOPE and NMP were performed through the portal vein and hepatic artery. RESULTS: From July 2016 to November 2017, we performed 7 HOPE procedures: 5 for DCD and 2 for DBD grafts. Two livers presented with macrovesicular steatosis >30% (1 DCD and 1 DBD). HOPE lasted 240 minutes (180-320 min) with a total ischemia time of 575 minutes (410-810 min). Six grafts were successfully transplanted. One DCD graft required additional evaluation using NMP. The graft was then discarded due to extensive hepatocellular necrosis. In the post-transplant course, acute and chronic renal failure were the main complications affecting 3 and 2 recipients, respectively. In our series, steatosis was the main risk factor for kidney injury. Patient and graft survival rate was 100% and no ischemic cholangiopathies were observed after 270 days (106-582 days). CONCLUSIONS: Our study confirms HOPE safety and efficacy for DCD and DBD grafts. These data are particularly significant for DCD management in the Italian setting where the mandatory 20-minute hands-off interval before death declaration further prolongs warm ischemia time.

Measurement of end-expiratory lung volume by oxygen washin-washout in controlled and assisted mechanically ventilated patients.

OBJECTIVE: Assessing limits of agreement with helium dilution and repeatability of a new system (lung funcution, LUFU) that measures end-expiratory lung volume (EELV) in mechanically ventilated patients using the O(2) washin (EELV(Win)) and washout (EELV(Wout)) technique. LUFU consists of an Evita 4 ventilator, a side-stream oxygen analyzer, and a dedicated PC software. DESIGN AND SETTING: Prospective human study in a general ICU of a University hospital. PATIENTS: Thirty-six mechanically ventilated patients. INTERVENTIONS: We obtained 36 couples of both EELV(Win) and EELV(Wout) measurements in each patient (5 with healthy lungs, 9 with ALI, 22 with ARDS). Measurements were obtained with patients ventilated either by assisted (ASB, 16 measurements) or controlled (CMV, 20 measurements) ventilation. In 19 of 20 cases in CMV, we obtained helium dilution measurements (EELV(He)). MEASUREMENTS AND RESULTS: Bias for agreement with EELV(He) was -16 +/- 156 and 8 +/- 161 ml, respectively, for EELV(Win) and EELV(Wout). Bias for agreement between EELV(Win) and EELV(Wout) was 28 +/- 78 and 23 +/- 168 ml, respectively, for CMV and ASB. During CMV bias for repeatability were 8 +/- 92 and 23 +/- 165 ml, respectively, for EELV(Win) and EELV(Wout). During ASB bias for repeatability were 32 +/- 160 and -15 +/- 147 ml, respectively, for EELV(Win) and EELV(Wout). CONCLUSIONS: The LUFU method showed good agreement with helium, and good repeatability during partial and controlled mechanical ventilation. The technique is simple and safe.

Depressed DHEA and increased sickness response behaviors in lame dairy cows with inflammatory foot lesions.

Lameness is a multifactorial condition influenced by the environment, genetics, management and nutrition. Detection of lameness is subjective and currently limited to visual locomotion observations which lack reliability and sensitivity. The objective of this study was to search for potential biomarkers of inflammatory foot lesions that underlie most cases of lameness in dairy cows, with a focus on the sickness response and relevant endocrine, immune and behavioral changes. Serum and peripheral blood mononuclear cells (PBMC) were collected from eight sound and eight lame high-producing Holstein cows. Immune cell activation was investigated in PBMCs using a candidate gene approach in which the expression of pro-opiomelanocortin, interleukin-1beta, l-selectin, matrix metalloproteinase-9 and glucocorticoid receptor-alpha was measured via quantitative real time-RT-PCR. Endocrine changes were investigated by monitoring serum concentrations of cortisol and dehydroepiandrosterone (DHEA). Additionally, systematic behavioral observations were carried out to characterize a behavioral profile associated with a sickness response typical of this condition. Lame cows showed significantly lower eating (P=0.01) and ruminating (P=0.01) behaviors and higher incidence of self-grooming (P=0.04) compared to sound cows. Lame cows also showed a 23% decrease in serum DHEA (P=0.01) and 65% higher cortisol:DHEA ratio (P=0.06) compared to sound cows. However, no significant differences were found in candidate gene expression between lame and sound cows. In association with sickness behaviors, serum DHEA concentration and cortisol:DHEA ratio are promising objective indicators of inflammatory foot lesions in dairy cattle and may be useful as diagnostic targets for animals in need of treatment.

Effects of early weaning and social isolation on the expression of glucocorticoid and mineralocorticoid receptor and 11beta-hydroxysteroid dehydrogenase 1 and 2 mRNAs in the frontal cortex and hippocampus of piglets.

Pigs weaned at young ages show more abnormal and aggressive behaviors and cognitive deficits compared to later weaned pigs. We investigated the effects of age, weaning and/or social isolation on the expression of genes regulating glucocorticoid response [glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11beta-hydroxysteroid dehydrogenases 1 and 2 (11beta-HSD1 and 11beta-HSD2)] in the frontal cortex and hippocampus. Early- (EW; n = 6) and conventionally-weaned (CW; n = 6) piglets were weaned at 10 and 21 days after birth, respectively. Non-weaned (NW) piglets of both ages (NW; n = 6/group) remained with their dams. Immediately before euthanasia, half of CW, EW and NW animals were socially isolated for 15 min at 12 (EW, NW) and 23 (CW, NW) days of age. Differences in amounts of 11beta-HSD1, 11beta-HSD2, GR and MR mRNA were determined by quantitative real-time RT-PCR and data subjected to multivariate linear mixed model analysis. When compared with NW piglets at 12 days of age, the hippocampi of EW piglets showed decreased gene expression (P < 0.01). Social isolation decreased gene expression (P < 0.05) in the frontal cortex of all piglets. Twelve-day-old piglets showed higher MR mRNA in the frontal cortex (P < 0.01) and lower 11beta-HSD2 and GR mRNA (P < 0.05) in the hippocampus compared to 23-day-old animals. Results indicate that EW affected the hippocampus of piglets at 12 days of age, while social isolation affected frontal cortex regardless of age. These results may be correlated with behavioral and cognitive changes reported in EW piglets.

Investigation of changes in global gene expression in the frontal cortex of early-weaned and socially isolated piglets using microarray and quantitative real-time RT-PCR.

We hypothesize that early-weaned piglets experience aberrant expression of stress-responsive genes in the frontal cortex, a key brain area involved in cognitive function and behavior organization. To test this hypothesis, female early-weaned piglets (EW; n = 6) were weaned 10 days after birth, while non-weaned piglets (NW; n = 6) were left with their dams. Half of EW (n = 3) and NW (n = 3) animals were socially isolated (SI) for 15 min at 12 days of age, when all animals (n = 12) were euthanized and tissue collected. The effects of EW and SI were examined by gene expression profiling using cDNA microarray hybridizations, generated from a porcine brain cDNA library. A total of 103 genes were differentially expressed (P < 0.05, fold change >1.25) among four direct comparisons. Forty-two genes had known functions, from which 24 showed relevant brain-related functions. Quantitative real-time polymerase chain reaction (Q-RT-PCR) was used to confirm regulation of expression of a subset of 6 genes with important brain functions, selected from the microarray outcomes. In non-weaned animals, a significant suppression of mRNA abundance for carboxypeptidase E, 14-3-3 protein and phosphoprotein enriched in astrocytes 15 kDa was observed in response to SI. Also, in early-weaned animals, diazepam binding inhibitor and actin-related protein 2/3 complex mRNA levels were suppressed in response to SI. Results suggest that social isolation of non- and early-weaned piglets may impact expression of genes involved in regulation of neuronal function, development, and protection in the frontal cortex of young pigs.

Chemical and biological characterization of dissolved organic matter from silver fir and beech forest soils.

Despite a growing attention to the dissolved organic matter (DOM) in terrestrial ecosystems and evidence of the fact that vegetation affects the quality of both undissolved and dissolved organic matter in soil, the role of DOM as a biological indicator is still poorly understood. In this work, the fertility of 59 sites, divided into eight key alliances of the order Fagetalia sylvaticae Pawl., was studied considering chemical and biological parameters such as soil DOM, hormone-like activity, low-molecular-weight (LMW) aliphatic and phenolic acids, and floristic data. Both non-parametric tests and principal component analysis (PCA) revealed differences between silver fir and beech forests and within each type of forest. There were also differences between neutrophilous and acidophilous types. What's more, PCA reveals the dominance of the auxin (IAA)-like activity, and of some phenolic acids in distinguishing the acidophilous beeches (ACI) form the other types, whereas the gibberellin (GA)-like activity is more relevant in neutrophilous conditions such as thermophilous (THE) and mesophilous (MESO) beeches and montane (MO), high montane (HMA), high montane (HMC) silver fir forests. The GA-like activity is also related to the succinic, fumaric, malonic, and l-malic acids in the MO, HMA and HMC silver fir forests. Moreover, the role of LMW aliphatic acids in mobilizing the hormone-like activity, which improves forest growth, is stressed. The growth of seedlings of Picea abies was influenced by the phenolic acid content. At concentrations between 1 and 100 microM, phenylacetic and protocatechuic acids inhibited root growth to the same extent as indoleacetic acid, while p-hydroxybenzoic acid had a stimulating effect comparable to that of gibberellic acid. The aliphatic and phenolic acids appear to be related to plant strategies that influence soil fertility affecting plant growth through rhizodeposition. The role of LMW aliphatic and phenolic acids as molecular markers of ecosystem function is noted.

Cell age-related monovalent cations content and density changes in stored human erythrocytes.

Conversion of erythrocyte membrane protein 4.1b to 4.1a occurs through a non-enzymatic deamidation reaction in most mammalian erythrocytes, with an in vivo half-life of approximately 41 days, making the 4.1a/4.1b ratio a useful index of red cell age [Inaba and Maede, Biochim. Biophys. Acta 944 (1988) 256-264]. Normal human erythrocytes distribute into subpopulations of increasing cell density and cell age when centrifuged in polyarabinogalactan density gradients. We have observed that, when erythrocytes were stored at 4 degrees C under standard blood bank conditions, the deamidation was virtually undetectable, as cells maintained the 4.1a/4.1b ratio they displayed at the onset of storage. By measuring the 4.1a/4.1b values in subpopulations of cells of different density at various time points during storage, a modification of the normal 'cell age/cell density' relationship was observed, as erythrocytes were affected by changes in cell volume in an age-dependent manner. This may stem from a different impact of storage on the imbalance of monovalent cations, Na(+) and K(+), in young and old erythrocytes, related to their different complement of cation transporters.

ICSH guidelines for the laboratory diagnosis of nonimmune hereditary red cell membrane disorders.

INTRODUCTION: Hereditary spherocytosis (HS), hereditary elliptocytosis (HE), and hereditary stomatocytosis (HSt) are inherited red cell disorders caused by defects in various membrane proteins. The heterogeneous clinical presentation, biochemical and genetic abnormalities in HS and HE have been well documented. The need to raise the awareness of HSt, albeit its much lower prevalence than HS, is due to the undesirable outcome of splenectomy in these patients. METHODS: The scope of this guideline is to identify the characteristic clinical features, the red cell parameters (including red cell morphology) for these red cell disorders associated, respectively, with defective cytoskeleton (HS and HE) and abnormal cation permeability in the lipid bilayer (HSt) of the red cell. The current screening tests for HS are described, and their limitations are highlighted. RESULTS: An appropriate diagnosis can often be made when the screening test result(s) is reviewed together with the patient's clinical/family history, blood count results, reticulocyte count, red cell morphology, and chemistry results. SDS-polyacrylamide gel electrophoresis of erythrocyte membrane proteins, monovalent cation flux measurement, and molecular analysis of membrane protein genes are specialist tests for further investigation. CONCLUSION: Specialist tests provide additional evidence in supporting the diagnosis and that will facilitate the management of the patient. In the case of a patient's clinical phenotype being more severe than the affected members within the immediate family, molecular testing of all family members is useful for confirming the diagnosis and allows an insight into the molecular basis of the abnormality such as a recessive mode of inheritance or a de novo mutation.