Organic cations in halide perovskite solid solutions: exploring beyond size effects.

Halide perovskites are a class of materials of consolidated optoelectronic and electrochemical applications, reaching efficiencies compared to established materials in respective fields. In this scenario, the design and understanding of composition-structure-property relations is imperative. In solid solutions containing mixed cations, some direct relations between the sizes of the substituents and the properties of perovskites are generally observed. However, in several cases, these relations are not observed, implying that other characteristics of these cations play a major role. Despite its importance, this understanding has not been comprehensively deepened. To address this issue, we synthesized and characterized the structure, electrical behavior, and stability of methylammonium lead iodide-based perovskites with equal amounts of the substituents guanidinium, ethylammonium, and acetamidinium. These three large organic cations have essentially equal sizes but other remarkably different characteristics, such as the number of N-H bonds, intrinsic dipole moment, and order of C-N bonds. Herein, we show that these cations have dramatically different effects over important fundamental and applied properties of resulting perovskites, including the orthorhombic-to-tetragonal and tetragonal-to-cubic phase transitions, microstructural development, ionic conductivity, I-V hysteresis, electronic carrier mobility, and stability against light-induced degradation. These effects are correlated with the characteristics of the large substituent cations and help pave the way for a better rational chemical design of halide perovskites.

Risk factors for purulent vaginal discharge and its association with reproductive performance of lactating Jersey cows.

Despite the rapid growth in popularity of the Jersey breed, most research on dairy cows in the United States has been done with the Holstein breed. Postpartum uterine diseases negatively influence reproductive performance of dairy cows and limited data are available regarding predisposing factors for uterine diseases in Jersey cows. Our objectives were to determine the prevalence and risk factors for purulent vaginal discharge (PVD) and its effect on fertility of lactating Jersey cows. This was a retrospective observational study with data collected from 3,822 Jersey cows. The Metricheck device was used for PVD diagnosis, and positive cases (≥50% of pus in exudate) were further classified for severity using the following 4 categories based on the amount of pus observed: 50 to 60% pus in exudate, 60 to 90% pus in exudate, 90 to 100% pus in exudate, and 90 to 100% pus in exudate + uterine fluid detected by palpation per rectum. Univariable and multivariable regression analyzes were conducted to dissect the risk factors for PVD and severity of PVD in Jerseys cows. The major risk factors for PVD were calving-related problems, retained fetal membrane, metritis, and days in the close-up diet. A subgroup of cows (n = 740) was scored for body condition and locomotion scores and had blood sampled in the peripartum for determination of plasma concentrations of nonesterified fatty acids and ß-hydroxybutyrate. In the multivariable analysis that considered data collected for the subgroup of cows, peripartum nonesterified fatty acids, postpartum ß-hydroxybutyrate, and peripartum locomotion and body condition scores were not retained in the reduced model of predictors of PVD. Not surprisingly, pregnancy per artificial insemination following the first and second services was reduced in cows diagnosed with PVD compared with cows without PVD. In addition, PVD was associated with increased odds of pregnancy loss after the first service and reduced hazard of pregnancy by 305 d in milk. The stratification of PVD severity according to the amount of pus observed and the consistency of the uterus was meaningful, as observed by the differences in reproductive outcomes between cows with diverging amounts of pus in the retrieved exudate. Risk factors for PVD in Jersey cows was similar to previously reported for Holstein cows, and a strong detrimental effect of PVD on fertility was also observed in Jersey cows.

Short communication: Simultaneous measurements of estrus behavior and plasma concentrations of estradiol during estrus in lactating and nonlactating dairy cows.

Estrus is an important behavior that can potentially be subjected to genomic selection. Circulating estradiol concentrations at estrus may be a useful phenotype if the absolute concentrations of estradiol are associated with overt phenotypes for estrus (activity, rump touches, or both; e.g., mounts, chinrests) that can be easily observed. The objective was to measure plasma estradiol concentrations at estrus and associate these measurements with the increase in activity (steps per hour) and rump touches received at estrus. We also tested the effect of lactation on the estrus traits that we measured. Cows (n = 11 lactating and n = 9 nonlactating) were treated with PGF2α to synchronize estrus. A jugular vein was cannulated to collect blood every 2 h for plasma estradiol measurement. Plasma LH was measured during the periestrual period to determine the time of the LH surge. Cows were fitted with an accelerometer to measure activity (steps per hour) and a capacitive touch sensing device to measure the number of rump touches and total touch time. Plasma estradiol concentrations were poorly correlated with overt signs of estrus during the period leading up to maximum estrus activity. After peak estrus activity (when cows were going out of estrus and plasma estradiol concentrations were decreasing), a stronger correlation was detected between overt signs of estrus and plasma estradiol concentrations. Effective selection for improved estrus expression based on plasma estradiol concentrations will depend on whether the cow is coming into or going out of estrus at the time of blood sampling. An association existed between lactation and fewer number of hours in estrus when estrus was defined by an increase in activity (steps per hour). Lactating cows had a shorter interval from the onset of estrus to the LH surge, and the shorter interval to the LH surge may have reduced the period of elevated estradiol during estrus in the lactating cows. Understanding mechanisms that control the sensitivity of the cow to estradiol and making appropriate selection decisions based on these mechanisms will likely increase overt signs of estrus in dairy cows.

Scanning electron microscopy of the surface epithelium of the bovine endometrium.

The surface epithelium of the bovine endometrium comprises at least 2 cell types (ciliated cells and secretory cells with microvilli), but their distribution and morphological changes over the estrous cycle are poorly understood. The objective was to quantify the number of ciliated cells and assess morphological changes in secretory cells on the uterine surface epithelium during the estrous cycle. Caruncular endometrium (CAR) and intercaruncular endometrium (ICAR) samples were collected from the uterine body, the horn ipsilateral to the corpus luteum or dominant follicle (H-CL/DF), and the horn contralateral to the corpus luteum or dominant follicle (H-NCL/NDF) from heifers following slaughter on d 0 (estrus; n = 5) or d 14 (mid-luteal phase; n = 5) of the estrous cycle. Samples were prepared for scanning electron microscopy at 1,000× magnification. Four to 10 fields (256 × 225 µm) for each sample were examined (n = 567 images). The number of ciliated cells was counted and the surface was scored for the morphology of the secretory cells (0 = absence of microvilli on surface; 3 = 100% of surface covered with microvilli). Ciliated cells were present in both the CAR and ICAR regions. The number of ciliated cells per field increased from d 0 to 14 in CAR and decreased from d 0 to14 in ICAR. The scanning electron microscopy revealed a general lack of uniformity in the lawn of microvilli on the surface of the endometrium. Based on the scores, approximately 25% of the fields had a surface that was <50% covered by microvilli. Depletion of microvilli may be explained by a normal process where apical protrusions are formed and either regress back into the cell surface or break to release their contents into the uterine lumen. These studies support the hypothesis that the surface of the luminal epithelium changes during the estrous cycle through a process that involves remodeling of the apical surface. The morphology of the apical surface may have a key role in governing pregnancy establishment.

Genome-wide association analysis and gene set enrichment analysis with SNP data identify genes associated with 305-day milk yield in Holstein dairy cows.

Milk production traits, such as 305-day milk yield (305MY), have been under direct selection to improve production in dairy cows. Over the past 50 years, the average milk yield has nearly doubled, and over 56% of the increase is attributable to genetic improvement. As such, additional improvements in milk yield are still possible as new loci are identified. The objectives of this study were to detect SNPs and gene sets associated with 305MY in order to identify new candidate genes contributing to variation in milk production. A population of 781 primiparous Holstein cows from six central Washington dairies with records of 305MY and energy corrected milk were used to perform a genome-wide association analysis (GWAA) using the Illumina BovineHD BeadChip (777 962 SNPs) to identify QTL associated with 305MY (P < 1.0 × 10-5 ). A gene set enrichment analysis with SNP data (GSEA-SNP) was performed to identify gene sets (normalized enrichment score > 3.0) and leading edge genes (LEGs) influencing 305MY. The GWAA identified three QTL comprising 34 SNPs and 30 positional candidate genes. In the GSEA-SNP, five gene sets with 58 unique and 24 shared LEGs contributed to 305MY. Identification of QTL and LEGs associated with 305MY can provide additional targets for genomic selection to continue to improve 305MY in dairy cattle.

Technical approaches to evaluate the surfactant-enhanced biodegradation of biodiesel and vegetable oils.

This research compared the effects of biosurfactant on the biodegradation of biodiesel and vegetable oils while validating two conceptually diverging methodologies. The two experimental setups were successfully modeled towards the effects of biosurfactants during biodegradation. We established the equivalence of both methodologies from the data output. As expected, the biosurfactants caused an increased oil uptake, thus increasing biodegradation performance. Cooking oils were favored by the microbial consortium as a carbon source when compared with biodiesel fuel, especially after use in food preparation. However, we found that biodiesel substrate standout with the highest biodegradation rates. Our results might indicate that a rapid metabolic change from the original compound initially favored biodiesels during the assimilation of organic carbon for a set specialized microbial inoculum. The data output was successfully combined with mathematical models and statistical tools to describe and predict the actual environmental behavior of biodiesel and vegetable oils. The models confirmed and predicted the biodegradation effectiveness with biosurfactants and estimated the required timeframe to achieve satisfactory contaminant removal.

A soluble inorganic pyrophosphatase from the cattle tick Rhipicephalus microplus capable of hydrolysing polyphosphates.

Polyphosphates have been found in all cell types examined to date and play diverse roles depending on the cell type. In eukaryotic organisms, polyphosphates have been investigated mainly in mammalian cells, and only a few studies have addressed arthropods. Pyrophosphatases have been shown to regulate polyphosphate metabolism. However, these studies were restricted to trypanosomatids. Here we focus on the tick Rhipicephalus microplus, a haematophagous ectoparasite that is highly harmful to cattle. We produced a recombinant R. microplus pyrophosphatase (rRmPPase) with the aim of investigating its kinetic parameters using polyphosphates as substrate. Molecular docking assays of RmPPase with polyphosphates were also carried out. The kinetic and Hill coefficient parameters indicated that rRmPPase has a greater affinity, higher catalytic efficiency and increased cooperativity for sodium phosphate glass type 15 (polyP15 ) than for sodium tripolyphosphate (polyP3 ). Through molecular docking, we found that polyP3 binds close to the Mg2+ atoms in the catalytic region of the protein, participating in their coordination network, whereas polyP15 interactions involve negatively charged phosphate groups and basic amino acid residues, such as Lys56, Arg58 and Lys193; polyP15 has a more favourable theoretical binding affinity than polyP3 , thus supporting the kinetic data. This study shows, for the first time in arthropods, a pyrophosphatase with polyphosphatase activity, suggesting its participation in polyphosphate metabolism.

Monocytes from male patients with ankylosing spondylitis display decreased osteoclastogenesis and decreased RANKL/OPG ratio.

The present study investigates the osteoclastogenic capacity of peripheral blood mononuclear cells (PBMCs) in male patients with ankylosing spondylitis (AS). We demonstrated that monocytes from these patients display a lower capacity to generate osteoclasts compared to cells from healthy controls, and osteoclastogenesis was negatively correlated with disease duration. INTRODUCTION: Ankylosing spondylitis (AS) is a disease characterized by new bone growth that leads to syndesmophyte formation but AS patients frequently present with low bone mineral density/fractures. Osteoclastogenesis in AS patients is poorly studied and controversial. The aim of this study is to determine if the osteoclastogenic capacity of PBMCs is different in AS patients compared to controls and the relationship between osteoclastogenesis and clinical/laboratory parameters. METHODS: PBMCs from 85 male AS patients and 59 controls were tested for CD16+ cells and induced to differentiate into osteoclasts over 3 weeks in vitro. Serum levels of RANKL, osteoprotegerin (OPG), C-terminal telopeptide of type I collagen (CTX), and amino-terminal pro-peptide of type I collagen (P1NP) were also evaluated. RESULTS: PBMCs from AS patients had fewer CD16+ cells and produced fewer osteoclasts compared to controls. Apoptosis occurred less frequently in osteoclasts obtained from AS patients than in osteoclasts from the controls. A lower RANKL/OPG and CTX/P1NP were observed in AS patients compared to controls. AS patients taking NSAIDs presented no difference regarding the number of OCs produced and the percentage of CD16+ cells compared to controls. However, patients taking TNF inhibitors (TNFi) presented lower OC numbers than controls. A negative correlation was demonstrated between the number of osteoclasts generated from PBMCs of AS patients and disease duration. CONCLUSION: Monocytes from male AS patients display a lower capacity to generate osteoclasts in vitro compared to cells from controls. Osteoclastogenesis was negatively correlated with disease duration. This finding supports the idea that osteoclasts play a role in the physiopathology of bone disease in AS patients.

Administration of Huperzia quadrifariata Extract, a Cholinesterase Inhibitory Alkaloid Mixture, has Neuroprotective Effects in a Rat Model of Cerebral Hypoxia-Ischemia.

Neonatal hypoxia-ischemia (HI) is an etiologic component of several neurologic pathologies associated to cognitive impairment. The mechanisms involved in HI-induced tissue damage start immediately after HI and extend for days. Acetylcholine is an important neurotransmitter in the central nervous system and exerts a protector effect on tissue damage by modulating inflammation, and cholinesterase inhibitors have shown neuroprotective properties and their action are often attributed to inhibition of the immune response. The administration of Huperzia quadrifariata alkaloid extract (HqAE), with potent and selective cholinesterase inhibitor properties, will reduce the HI induced behavioral deficits and tissue damage. A total of 84 newborn Wistar rat pups at post natal day 7 (PND7) were subjected to right carotid occlusion followed by 1 h of hypoxia (8% of O2) and i.p. injections of saline, vehicle or HqAE (10 mg/kg). Morris Water Maze and inhibitory avoidance tests were used to assess the cognitive function. Flow cytometry was performed at PND11. Histological analysis was performed at PND45. HqAE treatment was able to prevent the HI induced cognitive deficits in both tests and, at PND45, histological analysis showed that HqAE treatment reduced hippocampus tissue damage. Flow cytometry of the injured hippocampus revealed that the treatment was able to reduce cellular death and the number of infiltrating T cells. Altogether, these results show the therapeutic potential of the Huperzia quadrifariata alkaloid extract to prevent cognitive deficits and histological damage caused by neonatal hypoxia-ischemia, probably by reducing cellular death and T cell mobilization.

Induction of resistance of corn plants to Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) by application of silicon and gibberellic acid.

The aim of this study was to evaluate the effects of silicon application and administration of the phytohormone gibberellic acid on resistance of the corn plants to the fall armyworm (FAW), Spodoptera frugiperda, and their vegetative characteristics. We evaluated larval and pupal duration, survival and biomass, and adult longevity, malformation and fecundity of S. frugiperda after feeding on plant matter treated with silicon and/or gibberellic acid. The feeding preference of FAW first-instar larvae, the total leaf area consumed by the insects, and the vegetative parameters of corn plants were also evaluated. No significant differences were observed in the measured parameters of larval and pupal stages of S. frugiperda in response to silicon or gibberellic acid. In adult stage insects, the number of eggs per female was significantly reduced in insects derived from larvae fed plants treated with silicon or gibberellic acid. In a non-preference test, 48 h after release, caterpillars preferred control untreated plants and consumed less matter from plants that had received hormonal treatment (gibberellic acid). Gibberellic acid also altered the vegetative characteristics of plants, by increasing their height, shoot fresh and dry mass, and silicon content. We conclude that gibberellic acid can alter the vegetative characteristics and silicon uptake of corn plants, leading to a reduction in their consumption by S. frugiperda larvae and a decrease in female insect oviposition.

Ewes carrying the Booroola and Vacaria prolificacy alleles respond differently to ovulation induction with equine chorionic gonadotrophin.

The magnitude of ovulation rate (OR) after hormonal induction in sheep should be considered when prolific genotypes are used. We investigated for the first time the effect of the Vacaria allele and its combined effect with the Booroola prolificacy mutation on OR after hormonal treatment during breeding and anoestrous season. A hundred forty-nine Ile de France crossbred ewes, raised in natural pastures in South Brazil, were used to evaluate the OR after treatment with progestagen (MAP) followed or not by equine chorionic gonadotrophin (eCG) treatment (MAP + eCG). During the breeding season, 96% MAP-treated ewes ovulated in comparison to 97% of MAP + eCG-treated females. The double heterozygous carriers (BNVN) presented the higher OR, followed by the single Vacaria (NNVN) and Booroola (BNNN) heterozygous females and least the wild-type (NNNN) ewes. During anoestrus, 96% eCG-treated ewes ovulated, in contrast to 6% treated with MAP alone. The OR of the gonadotrophin-treated females was higher in BNVN and BNNN than NNVN and NNNN ewes. An additive effect in the OR of the two mutations was observed since OR in double heterozygous ewes was similar to the sum of the effects of the alleles of the single heterozygous carrier ewes.

Physicochemical evaluation of sheep milk yogurts containing different levels of inulin.

The present study aimed to evaluate the physicochemical parameters of sheep milk yogurt smoothies (SMY) containing inulin at different levels (0, 2, 4, and 6%). Titratable acidity and pH, yogurt bacteria counts, fatty acids profile, and healthy lipid indices were evaluated during 28 d of refrigerated storage. As expected for yogurts, Streptococcus thermophilus counts decreased 1 to 3 log cycles and Lactobacillus delbrueckii ssp. bulgaricus counts decreased 1 to 2 cycles from d 1 to 28. The protective effect of inulin on bacteria survival and viability in the food matrix was not verified in the prebiotic SMY during storage, regardless of inulin level. Although lower post-acidification was observed in prebiotic SMY due to inulin addition, no changes were verified in short-chain fatty acids (SCFA) or polyunsaturated fatty acids (PUFA). In contrast, an increase in medium- and long-chain fatty acids (MCFA and LCFA) and monounsaturated fatty acids (MUFA) was observed during storage in all SMY. The most significant levels of fatty acids in SMY were oleic acid, followed by palmitic and myristic acids. A high positive correlation between conjugated linoleic acid (CLA) and oleic acid (r=0.978) was observed. The cis-9,trans-11 CLA isomer represented approximately 78% of total PUFA and 2% of total fatty acids, whereas α-linoleic acid comprised about 22% PUFA and 1% of total fatty acids in SMY. The fatty acid changes during storage were associated with the metabolic activity of the starter bacteria, especially for oleic acid and cis-9,trans-11 CLA isomer. Thus, the SMY represented a great source of these compounds. We observed that inulin levels did not affect fatty acids. A nonsignificant decrease in atherogenic index was observed during storage in all SMY, and a positive correlation (r=0.973) was found between atherogenic index and thrombogenic index of SMY. High correlations were observed between lauric and myristic acids and saturated fatty acids (r=0.907 and r=0.894, respectively), providing evidence of their atherogenic and thrombogenic potential. A negative correlation was observed between stearic acid and atherogenic index (r=-0.612) and between oleic acid and atherogenic index. Sheep milk yogurt could be characterized as a food with low atherogenic and thrombogenic risk because of its healthy lipid composition. Therefore, addition of inulin to SMY could be a good option to improve functionality of this food matrix for dairy companies wishing to enter the functional food market.

Effects of recombinant bovine somatotropin during the periparturient period on innate and adaptive immune responses, systemic inflammation, and metabolism of dairy cows.

The aim of this experiment was to determine effects of treating peripartum dairy cows with body condition score ≥3.75 with recombinant bovine somatotropin (rbST) on immune, inflammatory, and metabolic responses. Holstein cows (253±1d of gestation) were assigned randomly to 1 of 3 treatments: untreated control (n=53), rbST87.5 (n=56; 87.5mg of rbST), and rbST125 (n=57; 125mg of rbST). Cows in the rbST87.5 and rbST125 treatments received rbST weekly from -21 to 28d relative to calving. Growth hormone, insulin-like growth factor 1, haptoglobin, tumor necrosis factor α, nonesterified fatty acids, ß-hydroxybutyrate, glucose, and cortisol concentrations were determined weekly from -21 to 21d relative to calving. Blood sampled weekly from -14 to 21d relative to calving was used for hemogram and polymorphonuclear leukocyte (PMNL) expression of adhesion molecules, phagocytosis, and oxidative burst. Cows were vaccinated with ovalbumin at -21, -7, and 7d relative to calving, and blood was collected weekly from -21 to 21d relative to calving to determine IgG anti-ovalbumin concentrations. A subsample of cows had liver biopsied -21, -7, and 7d relative to calving to determine total lipids, triglycerides, and glycogen content. Growth hormone concentrations prepartum (control=11.0±1.2, rbST87.5=14.1±1.2, rbST125=15.1±1.3ng/mL) and postpartum (control=14.4±1.1, rbST87.5=17.8±1.2, rbST125=21.8±1.1ng/mL) were highest for rbST125 cows. Cows treated with rbST had higher insulin-like growth factor 1 concentrations than control cows (control=110.5±4.5, rbST87.5=126.2±4.5, rbST125=127.2±4.5ng/mL) only prepartum. Intensity of L-selectin expression was higher for rbST125 than for control and rbST87.5 cows [control=3,590±270, rbST87.5=3,279±271, rbST125=4,371±279 geometric mean fluorescence intensity (GMFI)] in the prepartum period. The PMNL intensities of phagocytosis (control=3,131±130, rbST87.5=3,391±133, rbST125=3,673±137 GMFI) and oxidative burst (control=9,588±746, rbST87.5=11,238±761, rbST125=12,724±781 GMFI) were higher for rbST125 cows than for control cows during the prepartum period. Concentrations of serum IgG anti-ovalbumin tended to be higher for rbST125 cows than for control cows (control=0.75±0.11, rbST87.5=0.94±0.10, rbST125=1.11±0.11 optical density) in the prepartum period. Haptoglobin concentration was significantly reduced 7d postpartum for rbST125 treatment compared with control and rbST87.5 treatments (control=2.74±0.28, rbST87.5=2.81±0.28, rbST125=1.87±0.28 optical density). Although treatment tended to affect postpartum ß-hydroxybutyrate (control=747.5±40.2, rbST87.5=753.2±40.1, rbST125=648.8±39.7 µmol/L), it did not affect liver contents of total lipids, triglycerides, or glycogen. Incidence of metritis among rbST125 cows was reduced compared with that in control cows (control=23.1, rbST87.5=18.0, rbST125=7.8%). Treatment of dairy cows with 125mg of rbST improved innate immune responses and IgG concentration, with limited effects on metabolism.

Mutation in the protease cleavage site of GDF9 increases ovulation rate and litter size in heterozygous ewes and causes infertility in homozygous ewes.

Litter size (LS) in sheep is determined mainly by ovulation rate (OR). Several polymorphisms have been identified in the growth differentiation factor 9 (GDF9) gene that result in an increase in OR and prolificacy of sheep. Screening the databank of the Brazilian Sheep Breeders Association for triplet delivery, we identified flocks of prolific Ile de France ewes. After resequencing of GDF9, a point mutation (c.943C>T) was identified, resulting in a non-conservative amino acid change (p.Arg315Cys) in the cleavage site of the propeptide. This new allele was called Vacaria (FecG(v) ). A flock of half-sib ewes was evaluated for OR in the first three breeding seasons, and Vacaria heterozygotes had higher OR (P < 0.001), averaging 2.1 ± 0.1 when compared to 1.2 ± 0.1 in wild-type ewes. The OR was also influenced by age, increasing in the second and third breeding seasons (P < 0.001). In flocks segregating this allele, the LS was higher in mutant sheep (P < 0.001), averaging 1.61 ± 0.07 in heterozygotes and 1.29 ± 0.03 in wild-type ewes. Analysis of homozygote reproductive tract morphology revealed uterine and ovarian hypoplasia. Ovarian follicles continue to develop up to small antral stages, although with abnormal oocyte morphology and altered arrangement of granulosa cells. After the collapse of the oocyte in most follicles, the remaining cells formed clusters that persisted in the ovary. This SNP is useful to improve selection for dam prolificacy and also as a model to investigate GDF9 post-translation processing and the fate of the follicular cells that remain after the oocyte demise.

Prepartum stocking density: effects on metabolic, health, reproductive, and productive responses.

The objectives of the current experiment were to determine the effects of 2 prepartum stocking densities on milk yield, concentration of metabolites during the peripartum period, and health and reproductive parameters of dairy cows. Jersey cows enrolled in the experiment at 254±3 d of gestation were balanced for parity (nulliparous vs. parous) and previous lactation projected 305-d mature equivalent milk yield (parous) and assigned to 1 of 2 treatments: 80% headlock stocking density (80SD; 38 animals/48 headlocks) and 100% headlock stocking density (100SD; 48 animals/48 headlocks). The number of experimental units was 8 (4 replicates and 2 pens/treatment per replicate). In total, 154 nulliparous and 184 parous animals were enrolled in the 80SD treatment and 186 nulliparous and 232 parous animals were enrolled in the 100SD treatment. At the start of each replicate, treatments were switched within pen. Cows were milked thrice daily and monthly milk yield, fat and protein content, and somatic cell count data were recorded up to 155 d postpartum. Plasma nonesterified fatty acid concentration was measured weekly, from -18±3 to 17±3 d relative to calving, and plasma ß-hydroxybutyrate was measured weekly, from 1±2 to 17±3 d relative to calving. Cows were examined 1, 4±1, 7±1, 10±1, and 13±1 d relative to calving for diagnosis of uterine diseases. Blood was sampled for determination of progesterone concentration and resumption of ovarian cycles 35±3 and 45±3 d relative to calving. Average headlock (74.1±0.4 vs. 94.5±0.3%) and stall (80.8±0.4 vs. 103.1±0.4%) stocking density was lower for the 80SD treatment compared with the 100SD treatment. Treatment did not affect incidence of retained fetal membranes (80SD=5.1, 100SD=7.8%), metritis (80SD=21.2, 100SD=16.7%), acute metritis (80SD=9.9, 100SD=9.4%), and vaginal purulent discharge (80SD=5.8, 100SD=7.9%). Concentrations of nonesterified fatty acids (80SD=251.5±6.1, 100SD=245.9±5.6µmol/L) and ß-hydroxybutyrate (80SD=508.2±14.3, 100SD=490.9±13.6µmol/L) were not different between treatments. Treatment had no effect on percentage of cows removed from the herd on the first 60 d postpartum (80SD=6.1, 100SD=5.1%) and on rate of removal from the herd up to 305 d postpartum 80SD=referent, 100SD [adjusted hazard ratio (95% confidence interval)]=1.02 (0.75, 1.38). Percentages of cows pregnant to first (80SD=41.9, 100SD=48.4%) and second (80SD=49.3, 100SD=42.0%) postpartum AI were not different between treatments. Finally, treatment did not affect energy-corrected milk yield up to 155 d postpartum (80SD=33.8±0.5, 100SD=33.4±0.5kg/d). In herds with weekly or twice weekly movement of new cows to the prepartum pen and separate housing of nulliparous and parous animals, a target stocking density of 100% of headlocks on the day of movement is not expected to affect health, metabolic, reproductive, and productive parameters.

Effect of an Ovsynch56 protocol initiated at different intervals after insemination with or without a presynchronizing injection of gonadotropin-releasing hormone on fertility in lactating dairy cows.

The objectives of this study were to evaluate effects of 2 resynchronization protocols beginning at different intervals after artificial insemination (AI) on the pattern of return to estrus, ovarian responses, and pregnancy per AI (P/AI) to reinsemination. Lactating cows from 2 dairies, located in Texas (n=2,233) and Minnesota (n=3,077), were assigned to 1 of 4 timed AI (TAI) protocols 17 ± 3 d after AI. All cows were examined for pregnancy 31 ± 3 d after previous AI. Cows assigned to early Ovsynch56 (E-OV56) or OV56 received the Ovsynch56 protocol starting 24 or 31 d after AI, respectively. Cows assigned to early GnRH-GnRH-PGF(2α)-GnRH (E-GGPG) or GGPG received a presynchronizing GnRH injection 17 or 24 d after AI, respectively, 7 d before the start of the Ovsynch56 protocol. Cows observed in estrus after enrollment were inseminated on the same day. Ovaries were examined and blood was sampled for progesterone concentration on the day of first GnRH and PGF(2α) injection of the Ovsynch56 protocol. Pregnancy was diagnosed at 31 and 66 d after resynchronized AI. On the day of the first GnRH injection of the TAI, a higher percentage of cows on E-GGPG and GGPG protocols had a corpus luteum (E-GGPG=83.8, GGPG=91.2, E-OV56=80.4, and OV56=75.5%) and progesterone concentration >1 ng/mL (E-GGPG=62.5, GGPG=76.0, E-OV56=53.6, and OV56=60.8%) than cows assigned to other protocols. However, the percentage of cows ovulating to the first GnRH injection of TAI was not affected by treatment. Fewer E-GGPG and more OV56 cows were reinseminated in estrus (E-GGPG=23.7, GGPG=49.0, E-OV56=41.6, and OV56=57.6%). Treatment did not affect P/AI at 31 or 66 d for cows reinseminated in estrus. However, cows reinseminated in estrus had greater P/AI at 31 (40.0 vs. 27.5%) and 66 d (36.0 vs. 23.9%) than cows completing the TAI protocols. Among cows completing the TAI protocols, initiation of GGPG at 24 d after AI increased, whereas initiation of Ovsynch56 at 24 d after AI decreased P/AI at 31 d after reinsemination (E-GGPG=30.6, GGPG=28.3.0, E-OV56=22.3, and OV56=28.7%). Pregnancy per AI did not differ across treatment at 66 d after TAI (E-GGPG=26.6, GGPG=24.4, E-OV56=20.0, and OV56=24.1%). Overall, type of resynchronization protocol and protocol initiation time did not affect P/AI 66 d after reinsemination (E-GGPG=29.7, GGPG=30.5, E-OV56=26.1, and OV56=30.4%). In conclusion, GGPG resynchronization protocols and initiation of resynchronization protocol 24 d after AI reduced the number of cows reinseminated in estrus but neither the timing of initiation of resynchronization nor presynchronization with GnRH affected overall P/AI.

Histological assessment of non-ablative laser stimulation of tissue repair in acellular dermal grafts.

AIM: The objective of this study was to compare integration of AlloDerm® acellular dermal grafts in animals subjected to non-ablative laser irradiation and animals not exposed to this therapy. METHODS: Standardized AlloDerm® fragments measuring 5 mm² were grafted into the subcutaneous tissue overlying the calvaria in 32 Wistar rats. Laser therapy (685 ηm), at a dose of 4 J/cm2 per session, was applied immediately after surgical intervention and every 48 hours thereafter for a total of four applications. RESULTS: Analysis of histology slides revealed significantly greater edema in the control group. There was no neutrophil infiltration in the laser-irradiated group at any point during the study period, whereas such infiltration was present in control animals at three of the four points of observation. In the laser therapy group, lymphocyte infiltration was observed from day 1, whereas in the control group, it was only apparent from day 3. Vascularization was substantially greater in the control group. In the experimental group, the AlloDerm® graft was completely replaced by fibrous tissue. CONCLUSION: These findings suggest that add-on non-ablative laser therapy is an effective stimulator of healing and graft integration after placement of AlloDerm® acellular dermal grafts.

Immunogenicity and safety of two doses of a non-adjuvanted influenza A H1N1/2009 vaccine in young autoimmune rheumatic diseases patients.

OBJECTIVES: The aim of this study was to evaluate the immunogenicity and safety of the influenza A H1N1/2009 vaccine in children under 9 years old with autoimmune rheumatic diseases (ARD). METHODS: Thirty-eight ARD patients and 11 healthy children received two doses of non-adjuvanted influenza A/California/7/2009 (H1N1) virus-like vaccine. Subjects were evaluated before and 21 days after vaccination. Seroprotection (SP) and seroconversion (SC) rates, geometric mean titers (GMT) and factor increases (FI) in GMT were calculated. RESULTS: Mean ages were comparable between patients and controls. Pre-vaccination SP and GMT were similar in patients and controls (p > 0.05). Three weeks after immunization, SP (81.6% vs. 81.8%, p = 1.0), SC (81.6% vs. 90.9%, p = 0.66), GMT (151.5 vs. 282.1, p = 0.26) and the FI in GMT (16.7 vs. 36.3, p = 0.23) were similar in patients and controls, with both groups achieving an adequate response, according to the European Medicines Agency and Food and Drug Administration standards. Analysis of the possible factors influencing SC showed no difference in demographic data, leukocyte/lymphocyte counts or immunosuppressant use between seroconverted and non-seroconverted patients (p > 0.05). The vaccine demonstrated a satisfactory safety profile in this population. CONCLUSIONS: Two doses of influenza A H1N1/2009 vaccination induced an effective antibody response and caused adverse events in rare instances, suggesting this vaccine is appropriate and can be recommended for this age group.