Cow-related factors associated with intramammary infections during the 1st week postpartum in primiparous dairy cows in certified organic herds.

The objectives if this exploratory study were to identify variables associated with intramammary infections (IMI) during the 1st week of lactation in primiparous organic dairy cows, and to evaluate the association of those variables with somatic cell count (SCC) linear scores and milk yield in early lactation. Nulliparous cows (n = 240) were evaluated for: udder edema, teat edema, milk leakage (ML) and udder hygiene at weeks 6, 4, 2, and 1 prepartum and 1st week postpartum; body condition score (BCS) at 6 weeks prepartum and 1st week postpartum; age at calving (days), gestation length, dystocia, stillbirth, calf sex were included in the analysis. Milk samples from the 1st week postpartum were cultured using standard laboratory techniques and bacterial growth was considered IMI. Intramammary infection was observed in 58.7% of cows, with Staphylococcus chromogenes and S. aureus being the most prevalent pathogens. Cows with ML at 1st week postpartum were at 3.42 greater odds of IMI (P < 0.01). Cows with prepartum BCS ≥ 3.75 were at 3.12 greater odds of IMI (P < 0.01). Cows with ML in the 1st week of lactation had increased SCC (P < 0.01) and lower milk production in the second month of lactation (P = 0.05). Intervention studies are needed to evaluate if monitoring prepartum BCS and ML at early postpartum can aid in the control of IMIs in heifers in organic dairies.

Study of the immunologic response of marine-derived collagen and gelatin extracts for tissue engineering applications.

The host immunologic response to a specific material is a critical aspect when considering it for clinical implementation. Collagen and gelatin extracted from marine sources have been proposed as biomaterials for tissue engineering applications, but there is a lack of information in the literature about their immunogenicity. In this work, we evaluated the immune response to collagen and/or gelatin from blue shark and codfish, previously extracted and characterized. After endotoxin evaluation, bone marrow-derived macrophages were exposed to the materials and a panel of pro- and anti-inflammatory cytokines were evaluated both for protein quantification and gene expression. Then, the impact of those materials in the host was evaluated through peritoneal injection in C57BL/6 mice. The results suggested shark collagen as the less immunogenic material, inducing low expression of pro-inflammatory cytokines as well as inducible nitric oxide synthase (encoded by Nos2) and high expression of Arginase 1 (encoded by Arg1). Although shark gelatin appeared to be the material with higher pro-inflammatory expression, it also presents a high expression of IL-10 (anti-inflammatory cytokine) and Arginase (both markers for M2-like macrophages). When injected in the peritoneal cavity of mice, our materials demonstrated a transient recruitment of neutrophil, being almost non-existent after 24 hours of injection. Based on these findings, the studied collagenous materials can be considered interesting biomaterial candidates for regenerative medicine as they may induce an activation of the M2-like macrophage population, which is involved in suppressing the inflammatory processes promoting tissue remodeling. STATEMENT OF SIGNIFICANCE: Marine-origin biomaterials are emerging in the biomedical arena, namely the ones based in marine-derived collagen/gelatin proposed as cell templates for tissue regeneration. Nevertheless, although the major cause of implant rejection in clinical practice is the host's negative immune response, there is a lack of information in the literature about the immunological impact of these marine collagenous materials. This work aims to contribute with knowledge about the immunologic response to collagen/gelatin extracted from blue shark and codfish skins. The results demonstrated that despite some differences observed, all the materials can induce a macrophage phenotype related with anti-inflammation resolution and then act as immuno-modulators and anti-inflammatory inducible materials.

Effect of injectable trace mineral supplementation on peripheral polymorphonuclear leukocyte function, antioxidant enzymes, health, and performance in dairy cows in semi-arid conditions.

The objective of this study was to evaluate the effect of subcutaneous injections of 15 mg/mL Cu, 5 mg/mL Se, 60 mg/mL Zn, and 10 mg/mL Mn on health, performance, polymorphonuclear leukocyte (PMNL) function, circulating glutathione peroxidase (GPx) and superoxide dismutase (SOD) concentrations, and inflammation of dairy cows undergoing the transition period in high temperature-humidity index. A total of 923 multiparous cows from 2 commercial dairy farms were randomly allocated into 1 of 2 treatment groups as follows: control and injectable trace mineral supplementation (ITMS). Cows in the ITMS group received 7 mL of subcutaneous injections at dry-off (208 ± 3 d of gestation), 260 ± 3 d of gestation, and at 35 ± 3 d in milk (DIM). Data regarding health traits, reproductive performance, milk yield, and survivability were extracted from farm database software, and animals were followed-up until 300 DIM. For a subset of 142 cows from one herd, blood samples were collected at enrollment, and at 3 ± 1, 7 ± 1, 10 ± 1, and 35 ± 3 DIM to evaluate hematology, PMNL function, GPx and SOD concentrations, and circulating haptoglobin. Logistic regression was used to assess health and pregnancy per artificial insemination at first service. Cox proportional hazards models were used to evaluate hazard of pregnancy and culling. Mixed linear regression models accounting for repeated measures were used to assess all continuous variables collected over time. Parity, twinning, and previous gestation length were considered as potential confounders. Farm was included as a random effect. The ITMS cows tended to have lower incidence of metritis and stillbirth compared with control group. However, ITMS treatment did not influence the incidence of other diseases (e.g., mastitis, retained placenta), milk yield, reproductive performance, culling, and leukocyte count. Neutrophil-to-lymphocyte ratio, PMNL phagocytosis, and oxidative burst as well as intensity of the oxidative burst were greater for ITMS-treated cows in comparison to control cows. The ITMS cows had decreased expression of the adhesion molecule L-selectin on PMNL surface. The serum concentration of GPx and SOD were not affected by ITMS treatment. In conclusion, ITMS tended to reduce the incidence of metritis and stillbirth parturition, improved PMNL function, and improved the inflammatory status of dairy cows undergoing the transition period in high temperature-humidity index conditions. However, these findings did not translate into improved milk yield, reproductive performance, and survivability.

Prophylactic use of an ultra-diluted complex on health, metabolism, and performance of weaned Holstein calves and its carryover effect up to first lactation.

The objective of this study was to evaluate the effect of an ultra-diluted complex supplemented as a prophylactic strategy on the incidence risk of diseases, metabolism, and performance of weaned Holstein calves immediately after grouping. Additionally, the objective was to investigate the carryover effects on heifer-raising and first-lactation periods, in a double-blind, placebo-controlled trial. A total of 184 weaned Holstein calves (83 ± 7.9 d of life; 112.5 ± 11.7 kg of body weight; values are mean ± standard deviation) were allocated to 8 paddocks in a completely randomized block design experiment. During a 112-d period, animals received a total mixed ration and were randomly allocated into 2 groups: (1) CON: placebo-control (basal diet + calcium carbonate, top-dressed at 30 g/animal per day of placebo), or (2) UD: ultra-diluted complex (basal diet + TopVita-Real H, top-dressed at 30 g/animal per day of sulfur (10-60) + viola tricolor (10-14) + caladium seguinum (10-30) + zincum oxydatum (10-30) + phosphorus (10-60) + carduus marianus (10-60) + colibacillinum (10-30) + podophyllum (10-30) + vehicle: calcium carbonate; quantum sufficit 1 kg). Incidence risk of diseases, number of days with disease, and mortality events were recorded daily. Performance measurements and blood collection were carried out at enrollment and every 28 d until the end of the study period. The incidence risk of digestive problems was higher for the CON group. Additionally, the CON group had higher cumulative number of days per calf affected by tick-borne disease. Blood analysis revealed that CON calves had increased mean corpuscular hemoglobin amount and concentration than UD calves on d 28 after enrollment and had increased serum albumin and total protein concentration on d 84. Furthermore, higher albumin to globulin ratio was found in CON calves than in those of UD group at enrollment; however, on d 56, the inverse was observed. On d 28 after enrollment, CON calves had increased serum cortisol concentration. Regarding body measurements, higher withers height and body depth were detected in UD calves, on d 28 and d 84, respectively. Finally, higher culling risk was observed in the CON group during the heifer-raising period. In conclusion, a lower incidence risk of digestive problems and days with tick-borne disease were detected in the UD group. Additionally, systemic inflammation was improved by UD complex based on circulating inflammation and stress biomarkers on specific days after enrollment. However, UD product did not improve performance at the post-weaning, heifer-raising, and first-lactation periods.

Associations between circulating levels of natural antibodies, total serum immunoglobulins, and polymorphonuclear leukocyte function in early postpartum dairy cows.

The objective of this study was to investigate the associations between natural antibodies (NAbs) and total serum immunoglobulins (Igs) concentrations (both from IgG and IgM isotypes), with variables of polymorphonuclear leukocytes (PMN) function of dairy cows in the early postpartum period. Seventy-five healthy postpartum dairy cows at 2 ± 1 days in milk were enrolled in a cross-sectional study. Natural and total antibodies in serum samples were measured by ELISA. Flow cytometry was used to determine the phagocytosis and oxidative burst capacities of PMN and the quantification of the adhesion molecule l-selectin. Leukocyte count and differentials, and serum haptoglobin were also measured. A positive correlation between NAbsIgM and total serum IgM and between NAbsIgM and NAbsIgG were found. Before performing the associations between circulating Igs concentrations and PMN function variables, cows were categorized into having low, medium or high circulating antibodies based on their NAbs and total Igs serum concentrations. None of the PMN variables assessed differed between low, medium, and high cows for both NAbsIgM and NAbsIgG. While associations between PMNs function and total IgG were not observed, some associations between total IgM concentrations and PMN activity were found. Cows with high serum IgM had greater phagocytic activity compared to cows in the low IgM group. Finally, the proportion of PMN that performed oxidative burst and PMN surface expression of l-selectin intensity was greater in high total IgM group when compared to medium and low groups. In conclusion, association between NAbs, and PMN activity variables were not observed, but total serum IgM was associated to some PMN function variables in early post-partum dairy cows.

Adaptive immunity in the postpartum uterus: Potential use of vaccines to control metritis.

After parturition, dairy cows rely on an effective innate immune response, through the actions of neutrophils, macrophages, and antimicrobial peptides, to clear the uterus from pathogenic bacteria, such as E. coli, Bacteroides spp, F. necrophorum and T. pyogenes. However, the role of adaptive immunity in the postpartum uterus is less understood. In this review, we explore concepts of mucosal adaptive immunity and discuss recent findings regarding the efficacy of vaccines to reduce metritis in dairy cows. Areas of lymphocytic aggregates are seen throughout the bovine reproductive tract after parturition, but it is unknown if their development is influenced by previous exposure to pathogens or other intrinsic factors. Through the actions of Treg cells and γδ T cells, the uterus is an immune-tolerant environment during pregnancy. After parturition, the dynamics in the endometrial and circulating lymphocytic populations differ among cows that develop uterine diseases and healthy counterparts. However, the functionality of those cells has not yet been determined. It has been hypothesized that cows that fail to switch their uterine environment from an anti-inflammatory state prior to parturition to a pro-inflammatory state after calving are more susceptible to uterine infections. Given the nature of metritis related pathogens and the importance of innate immunity to uterine defense mechanisms, we speculate that an adaptive immunity biased towards a Th1/Th17 cellular response will provide best protection against uterine infections. Few studies have evaluated the efficacy of immunization in reducing the incidence of metritis in dairy cows revealing inconsistent findings.

Collagen-based bioinks for hard tissue engineering applications: a comprehensive review.

In the last few years, additive manufacturing (AM) has been gaining great interest in the fabrication of complex structures for soft-to-hard tissues regeneration, with tailored porosity, and boosted structural, mechanical, and biological properties. 3D printing is one of the most known AM techniques in the field of biofabrication of tissues and organs. This technique opened up opportunities over the conventional ones, with the capability of creating replicable, customized, and functional structures that can ultimately promote effectively different tissues regeneration. The uppermost component of 3D printing is the bioink, i.e. a mixture of biomaterials that can also been laden with different cell types, and bioactive molecules. Important factors of the fabrication process include printing fidelity, stability, time, shear-thinning properties, mechanical strength and elasticity, as well as cell encapsulation and cell-compatible conditions. Collagen-based materials have been recognized as a promising choice to accomplish an ideal mimetic bioink for regeneration of several tissues with high cell-activating properties. This review presents the state-of-art of the current achievements on 3D printing using collagen-based materials for hard tissue engineering, particularly on the development of scaffolds for bone and cartilage repair/regeneration. The ultimate aim is to shed light on the requirements to successfully print collagen-based inks and the most relevant properties exhibited by the so fabricated scaffolds. In this regard, the adequate bioprinting parameters are addressed, as well as the main materials properties, namely physicochemical and mechanical properties, cell compatibility and commercial availability, covering hydrogels, microcarriers and decellularized matrix components. Furthermore, the fabrication of these bioinks with and without cells used in inkjet printing, laser-assisted printing, and direct in writing technologies are also overviewed. Finally, some future perspectives of novel bioinks are given.

Peripheral Nerve Injury: Current Challenges, Conventional Treatment Approaches, and New Trends in Biomaterials-Based Regenerative Strategies.

Damage to peripheral nerves is a widely extended health problem, causing important socioeconomic costs worldwide. Indeed, peripheral nerve injuries (PNI) have been concerning the medical community for many decades. Nevertheless, despite the increase in knowledge in the injury physiopathology and the great research efforts being undertaken, the current standard grafting strategies used to repair PNI are not as efficient as desired. Although alternative engineered nerve grafts are already commercialized, their clinical performance is suboptimal. In this review, a general description of the circumstances and repercussions surrounding the PNI pathological state are presented, together with the treatment limitations and current challenges when addressing both short- and long-gap defects. In addition, potential therapeutic molecules are considered, while innovative regenerative strategies have been identified. Finally, the most relevant advances on the use of a wide range of biomaterials for the development of novel medical devices are also overviewed in depth, considering strategies making use of either empty or filled nerve conduits for guided tissue regeneration.

Short communication: Partial replacement of ground corn with algae meal in a dairy cow diet: Milk yield and composition, nutrient digestibility, and metabolic profile.

This study was undertaken to evaluate the effects of partially replacing dietary ground corn with a microalgae meal from Prototheca moriformis (composed of deoiled microalgae and soyhulls) on milk yield and composition, nutrient intake, total-tract apparent digestibility, and blood profile of lactating dairy cows. Twenty multiparous Holstein cows (57.7±49.4d in milk, 25.3±5.3 of milk yield, and 590±71kg of live weight at the start of experiment, mean ± standard deviation) were used in a cross-over design experiment, with 21-d periods. Diets were no microalgae meal (CON) or 91.8g/kg of microalgae meal partially replacing dietary ground corn (ALG). Cows showed similar milk yield and composition. The 3.5% fat-corrected milk production was 30.2±1.34kg/d for CON and 31.1±1.42kg/d for ALG. Despite cows having similar dry matter intake, ALG increased neutral detergent fiber and ether extract intake. In addition, cows fed ALG exhibited higher ether extract digestibility. No differences were detected in glucose, urea, amino-aspartate transferase, and gamma-glutamyl transferase blood concentrations. Feeding ALG increased the total cholesterol and high-density lipoprotein in blood compared with CON. The microalgae meal may partially replace ground corn in diets of lactating cows without impairing the animal's performance.

Revealing the potential of squid chitosan-based structures for biomedical applications.

In recent years, much attention has been given to different marine organisms, namely as potential sources of valuable materials with a vast range of properties and characteristics. In this work, ß-chitin was isolated from the endoskeleton of the giant squid Dosidicus gigas and further deacetylated to produce chitosan. Then, the squid chitosan was processed into membranes and scaffolds using solvent casting and freeze-drying, respectively, to assess their potential biomedical application. The developed membranes have shown to be stiffer and less hydrophobic than those obtained with commercial chitosan. On the other hand, the morphological characterization of the developed scaffolds, by SEM and micro-computed tomography, revealed that the matrices were formed with a lamellar structure. The findings also indicated that the treatment with ethanol prior to neutralization with sodium hydroxide caused the formation of larger pores and loss of some lamellar features. The in vitro cell culture study has shown that all chitosan scaffolds exhibited a non-cytotoxic effect over the mouse fibroblast-like cell line, L929 cells. Thus, chitosan produced from the endoskeletons of the giant squid Dosidicus gigas has proven to be a valuable alternative to existing commercial materials when considering its use as biomaterial.

Origins of the middle meningeal artery and its probableembryological mechanism: a review

The middle meningeal artery is traditionally described by the human anatomy books as having its origin inthe first portion of the maxillary artery. However, more detailed studies have shown that there are numerousvariations on this arterial segment with considerable clinical importance. Therefore, the aim of this study wasto perform a wide review about this aspect analyzing scientific articles and textbooks. Thus, this review shallbe useful for clinicians, surgeons and academics that manipulate and keep interest for this particular anatomicalsite.

Ossification of styloid ligament and its clinical implication a report of human casses

A case of ossification of the right styloid ligament in an adult human caucasian skull was verified duringroutine activities of the Laboratory of Human Anatomy at the University of Santa Cruz do Sul - Brazil. Thisstudy presents its clinical considerations and applications. The presence of other anatomical variations was notverified.

Conformational analysis of the antimalarial agent quinidine.

Quinidine is an active antimalarial compound extracted from the bark of Cinchona trees. The activity differences among structurally related molecules appears to depend on the absolute stereochemistry of some functional groups, a result that stimulated a detailed conformational analysis of these molecules of biological interest. In the present study, the potential energy surface (PES) for the antimalarial agent quinidine (C20H24O2N2) has been comprehensively investigated using the molecular mechanics (MM) and quantum mechanical semiempirical AM1 and PM3 methods. Six distinct minimum energy conformations were located on the multidimensional PES and also characterized as true minima through harmonic frequency analysis. The relative stabilities and thermodynamic properties are reported. The coexistence of different conformers is discussed for the first time in the literature based on the transition state (TS) structures located on the PES for the quinidine molecule. The theoretical results reported in the present study are in agreement with the experimental proposal, based on NMR data, that there are two conformations existing in solution for the quinidine molecule.

Quaternary ammonium salt derivatives of allylphenols with peripheral analgesic effect.

Ammonium salt derivatives of natural allylphenols were synthesized with the purpose of obtaining potential peripheral analgesics. These drugs, by virtue of their physicochemical properties, would not be able to cross the blood brain barrier. Their inability to enter into the central nervous system (CNS) should prevent several adverse effects observed with classical opiate analgesics (Ferreira et al., 1984). Eugenol (1) O-methyleugenol (5) and safrole (9) were submitted to nitration, reduction and permethylation, leading to the ammonium salts 4, 8 and 12. Another strategy applied to eugenol (1), consisting in its conversion to a glycidic ether (13), opening the epoxide ring with secondary amines and methylation, led to the ammonium salts 16 and 17. All these ammonium salts showed significant peripheral analgesic action, in modified version of the Randall-Sellito test (Ferreira et al., 1978), at non-lethal doses. The ammonium salt 8 showed an activity comparable to that of methylnalorphinium, the prototype of an ideal peripheral analgesic (Ferreira et al., 1984).

Quaternary ammonium salt derivatives of allylphenols with peripheral analgesic effect

Ammonium salt derivatives of natural allylphenols were synthesized with the purpose of obtaining potential peripheral analgesics. These drugs, by virtue of their physicochemical properties, would not be able to cross the blood brain barrier. Their inability to enter into the central nervous system (CNS) should prevent several adverse effects observed with classical opiate analgesics (Ferreira et al., 1984). Eugenol (1) O-methyleugenol (5) and safrole (9) were submitted to nitration, reduction and permethylation, leading to the ammonium salts 4, 8 and 12. Another strategy applied to eugenol (1), consisting in its conversion to a glycidic ether (13), opening the epoxide ring with secondary amines and methylation, led to the ammonium salts 16 and 17. All these ammonium salts showed significant peripheral analgesic action, in modified version of the Randall-Sellito test (Ferreira et al. 1978), at non-lethal doses. The ammonium salt 8 showed an activity comparable to that of methylnalorphinium, the prototype of an ideal peripheral analgesic (Ferreira et al., 1984).