Mastitis represents an important health problem in dairy sheep and somatic cell count (SCC) is frequently used as indicator of subclinical intra-mammary infection. Cytokines might represent another reliable and sensitive tool for defining the inflammatory status in relation with the SCC, thus, the levels of 12 key immune cytokines were monitored in ovine milk samples with different SCC values. First, samples were divided into five SCC-groups: group 1 (0–300 cell/mL*1000, n = 16), group 2 (300–500 cell/mL*1000, n = 16), group 3 (500–1,000 cell/mL*1000, n = 16), group 4 (1000–2000 cell/mL*1000, n = 15), group 5 (> 2000 cell/mL*1000, n = 16). Samples belonging to group 5 presented the highest values of IL-1α, IL-β, IL-6, MIP-1α, MIP-1β, IFN-γ, IL-17, IL-10. Samples belonging to group 1 presented IL-4 levels higher than to those belonging to groups 2–3-4, and lower IL-1α and MIP-1α values compared to groups 3–4. Next, cluster analysis was performed. Three clusters were defined: cluster 1 (samples with low SCC values negative to bacterial culture), cluster 2 (samples positive to culture isolation with intermediate SCC values), cluster 3 (samples positive to culture isolation and with high SCC levels). Samples in cluster 1 were characterised by low levels of IFN-γ, IL-1α, IL-1β, IL-6, MIP-1α, whereas samples in cluster 3 presented the highest values of these five cytokines. Samples in cluster 2 presented higher levels of IL-1α and MIP-1α compared to cluster 1, as well as lower levels of IL-4. MIP-1α and IL-1α showed the strongest correlation with SCC values. Overall, our data revealed that intra-mammary infection in dairy sheep was correlated with high levels of pro-inflammatory cytokines in milk samples, which reflect the presence of an inflammatory reaction and confirm the utility of SCC determination in the diagnosis of subclinical mastitis.

BackgroundStaphylococcal mastitis is a common disease of small ruminants causing major economic losses. The problem is particularly significant in the rural areas of the Mediterranean region, where almost two thirds of the global sheep and a quarter of the global goat milk are produced. This study aimed to gain insight into the genotypes, antimicrobial resistance (AMR) profiles and virulence factors of Staphylococcus aureus isolated from clinical mastitis in small ruminants’ farms from different 25 dairy herds in six different provinces in Sardinia, Italy between December 2021 and May 2022.MethodsThirty- two S. aureus were phenotypically identified and confirmed by real-time PCR. Whole-genome sequencing (WGS) was conducted and the sequence data were analyzed to reveal the genetic diversity, AMR markers and virulence genes to draw a conclusion for a current situation of small ruminants’ clinical mastitis in dairy herds in the region and the potential public health risk. Furthermore, the phylogenetic relations between S. aureus strains within one farm and from various farms was analyzed.ResultsAll isolates proved to be phenotypically Methicillin-susceptible S. aureus (MSSA), and none of them harbored mecA/C genes. The antimicrobial resistance against Tetracycline and Erythromycin were 15.62% and 3.12%, respectively. The isolates were assigned to ten sequence types in addition to five different clonal complexes using multilocus sequence typing (MLST). Sequence types ST133 (46.9%) and ST700 (21.9%) were the dominant types, and the majority of isolates were assigned to either CC133 (56.25%) or CC130 (34.4%). Twelve different spa-types were identified among isolates, while six isolates were not assigned to known spa-types. The dominant spa-type was t1773 (18.75%) which is known to be associated with CC130. All Tetracycline-resistant isolates harbored tet genes. The only Erythromycin-resistant isolate carried the erm(T) gene. The leucocidin genes lukF-PV (P83)/lukM were detected in 20 isolates (62.5%), while one isolate (ST522) carried a chimeric leukocidin.ConclusionIn conclusion, this study showed a considerable genetic diversity of S. aureus isolated from sheep and goat mastitis in Sardinia, Italy, and the prominent sensitivity to most of antimicrobial agents relevant for mastitis treatment. These findings inform about the local mastitis control strategies and highlight a low immediate public health risk from antimicrobial resistance in this setting.

ORF virus (ORFV) causes contagious ecthyma ("ORF"), a disease of sheep and goats characterized by lesions ranging from vesicles and pustules to atypical papilloma-like and angiomatous lesions in the skin and mucosae. The authors investigated the molecular factors leading to the ORF-associated atypical tumor-like changes. Fifteen lambs, 15 kids, and an adult ram clinically affected by natural ORFV infection were enrolled in the study and examined by several methods. ORFV was detected by viral culture or real-time polymerase chain reaction (RT-PCR) in the lesioned tissues and in the blood of the clinically affected sheep and goats. Surprisingly, ORFV was also detected in the blood of healthy goats from an affected herd. Microscopically, they found a pseudo-papillomatous proliferation of the epithelium, while the dermis and lamina propria were expanded by a proliferating neovascular component that highly expressed the viral vascular endothelial growth factor (vVEGF) and its host receptor vascular endothelial growth factor receptor 2 (VEGFR2). Immunohistochemistry, immunofluorescence, and in situ hybridization for mRNA showed that epidermal growth factor receptor (EGFR) was expressed in the fibrovascular component, in the infiltrating CD163+ macrophages, and in the basal stratum of the epidermis. Confocal immunofluorescence microscopy demonstrated that CD163+ macrophages were associated with VEGF and VEGFR2. Finally, they found by quantitative RT-PCR the overexpression of the interleukin-6 and VEGFR2 genes in the lesioned tissues. These findings suggest that ORFV activates an inflammatory reaction characterized by CD163+ macrophages expressing EGFR and VEGFR2, which might play an oncogenic role through synergistic action with vVEGF signaling.

An LC–Orbitrap–HRMS method was developed and validated for the simultaneous determination of four tetracyclines—oxytetracycline (OTC), tetracycline (TC), doxycycline (DC), and chlortetracycline (CTC)—in milk. This method involves sample extraction with McIlvaine–EDTA buffer solution (pH 4) and solid-phase extraction (SPE) with Oasis HLB cartridges, followed by the evaporation of the extract and its reconstitution with a 14% methanol aqueous solution before injection into the instrumental system. This method has been validated in terms of linearity, sensitivity, selectivity, precision, and accuracy, in accordance with Commission Decision 2002/657/EC requirements. Compared to existing methods, this approach optimally combines a quantitative procedure for extracting analytes from the milk of different species, including sheep, bovines, and goats, with a very short LC–Orbitrap–HRMS instrumental analysis time (only 8 min), simultaneously ensuring high precision, sensitivity, and applicability as a rapid confirmation method in official food control laboratories. The proposed method was applied to determine the concentration levels of OTC in milk samples derived from healthy Sardinian sheep and those naturally infected with Streptococcus uberis, after the intramuscular administration of an antibiotic, in order to evaluate how much of the drug was “subtracted” during penetration from blood into milk, with a potential effect of reducing its therapeutic efficacy.

Mastitis is one of the most impacting diseases in dairy farming, and its sensitive and specific detection is therefore of the greatest importance. The clinical evaluation of udder and mammary secretions is typically combined with the milk Somatic Cell Count (SCC) and often accompanied by its bacteriological culture to identify the causative microorganism. In a constant search for improvement, several non-enzymatic milk proteins, including milk amyloid A (M-SAA), haptoglobin (HP), cathelicidin (CATH), and lactoferrin (LF), have been investigated as alternative biomarkers of mastitis for their relationship with mammary gland inflammation, and immunoassay techniques have been developed for detection with varying degrees of success. To provide a general overview of their implementation in the different dairy species, we carried out a systematic review of the scientific literature using the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines. Our review question falls within the type “Diagnostic test accuracy questions” and aims at answering the diagnostic question: “ Which are the diagnostic performances of mastitis protein biomarkers investigated by immunoassays in ruminant milk?” . Based on 13 keywords combined into 42 searches, 523 manuscripts were extracted from three scientific databases. Of these, 33 passed the duplicate removal, title, abstract, and full-text screening for conformity to the review question and document type: 78.8% investigated cows, 12.1% sheep, 9.1% goats, and 6.1% buffaloes (some included more than one dairy species). The most frequently mentioned protein was M-SAA (48.5%), followed by HP (27.3%), CATH (24.2%) and LF (21.2%). However, the large amount of heterogeneity among studies in terms of animal selection criteria (45.5%), index test (87.9%), and standard reference test (27.3%) resulted in a collection of data not amenable to meta-analysis, a common finding illustrating how important it is for case definitions and other criteria to be standardized between studies. Therefore, results are presented according to the SWiM (Synthesis Without Meta-analysis) guidelines. We summarize the main findings reported in the 33 selected articles for the different markers and report their results in form of comparative tables including sample selection criteria, marker values, and diagnostic performances, where available. Finally, we report the study limitations and bias assessment findings.

Rationale: Although platelets are considered key inflammatory mediators in respiratory diseases, their role in bronchiectasis has not been fully explored. Objectives: We hypothesized that thrombocytosis in stable state may be associated with bronchiectasis severity and worse clinical outcomes. Methods: Patients with bronchiectasis have been enrolled from 10 centers in Europe and Israel, with platelet count recorded during stable state. The primary outcome was 5-year all-cause mortality. Secondary outcomes included exacerbations, hospitalizations, and mortality at 1, 2, and 3-year follow-up. Analyses were conducted using logistic regression after adjustment for confounding variables. Results: Among the 1,771 patients (median age, 67 yr; 63.4% female) included, 136 (7.7%) had thrombocytosis. Patients with thrombocytosis had a significantly higher disease severity, worse quality of life, higher number of exacerbations and hospitalizations, and higher mortality rate at both 3-year (23 [22.8%] vs. 83 [8.5%], respectively; P < 0.01) and 5-year (26 [35.1%] vs. 116 [15.9%], respectively; P < 0.01) in comparison with those with normal platelet count. Thrombocytosis was significantly associated with hospitalizations because of severe exacerbations (odds ratio [OR], 1.83; 95% confidence interval [CI], 1.20-2.79; P = 0.01) after 1-year follow-up, as well as increased 3-year (OR, 3.06; 95% CI, 1.74-5.39; P < 0.01) and 5-year (OR, 2.46; 95% CI, 1.39-4.37; P < 0.01) mortality. Conclusions: Platelets represent a cheap and easy-to-evaluate biomarker, and the presence of thrombocytosis during stable state is associated with disease severity, hospitalizations because of exacerbations, poor quality of life, and mortality in adults with bronchiectasis.

Objective The aim of this study was to compare the resulting short-term complications after simultaneous (SIM) or staged (ST) tibial plateau levelling osteotomy (TPLO) in patients that presented with bilaterally torn cranial cruciate ligaments. Study Design This was a retrospective study. Materials and Methods Medical records of dogs diagnosed with bilateral cranial cruciate ligament disease that underwent bilateral SIM or ST TPLO surgery with arthroscopy (2005-2015) were reviewed to evaluate the intra- and postoperative complications. Data were analysed and major and minor complications were compared between the two groups. Results A total of 176 client-owned dogs (352 stifles) that had TPLO performed bilaterally with SIM or ST procedures were included for analysis. The overall complication rate was 47.5% for the SIM group and 19.5% for the ST group. The incidence of major complications was 10.1 and 3.8% in the SIM and ST groups respectively. Minor complications were 38.4 and 15.6% in the SIM and ST groups respectively. Tibial tuberosity fractures requiring revision were noted in 2% of the SIM group and none of the ST group. Clinical Significance Although no significant difference was noted in major complication rate or tibial tuberosity fractures in our cohort of patients, a Type II statistical error was found. The complication rate in the SIM group is numerically greater by a factor of 2-3x or more for nearly all categories. Surgeons should consider the risks and benefits of SIM versus ST procedures as well as relevant individual patient and client factors before electing either alternative.

Late lactation is a critical moment for making mastitis management decisions, but in small ruminants the reliability of diagnostic tests is typically lower at this stage. We evaluated somatic cell counts (SCC) and cathelicidins (CATH) in late lactation sheep and goat milk for their relationship with intramammary infections (IMI), as diagnosed by bacteriological culture (BC). A total of 315 sheep and 223 goat half-udder milk samples collected in the last month of lactation were included in the study. IMI prevalence was 10.79% and 15.25%, respectively, and non-aureus staphylococci were the most common finding. Taking BC as a reference, the diagnostic performance of SCC and CATH was quite different in the two species. In sheep, receiver operating characteristic (ROC) analysis produced a higher area under the curve (AUC) value for CATH than SCC (0.9041 versus 0.8829, respectively). Accordingly, CATH demonstrated a higher specificity than SCC (82.92% versus 73.67%, respectively) at comparable sensitivity (91.18%). Therefore, CATH showed a markedly superior diagnostic performance than SCC in late lactation sheep milk. In goats, AUC was <0.67 for both parameters, and CATH was less specific than SCC (61.90% versus 65.08%) at comparable sensitivity (64.71%). Therefore, both CATH and SCC performed poorly in late lactation goats. In conclusion, sheep can be screened for mastitis at the end of lactation, while goats should preferably be tested at peak lactation. In late lactation sheep, CATH should be preferred over SCC for its higher specificity, but careful cost/benefit evaluations will have to be made.

Little information is available on the influence of sex in combination with smoking habits and combined oral contraceptives (COC) use on cellular inflammatory indexes such as neutrophil/lymphocyte ratio (NLR), derived NRL (dNLR), platelet/lymphocyte ratio (PLR), monocyte/lymphocyte ratio (MLR), mean platelet volume/platelet count (MPV/PLT), aggregate inflammation systemic index (AISI), and systemic inflammation response index (SIRI), which are cost-effective biomarkers to assessing inflammation. Therefore, the effect of COC was studied alone or in association with smoking and compared with results from healthy COC-free women and men. Furthermore, the association of cellular inflammatory indexes with endothelial function (arginine (Arg), asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA) and lipid peroxidation (malondialdehyde MDA) biomarkers was evaluated. Blood was collected for hematological and biochemical analysis, which were used to calculate PLR, NLR, dNLR, MLR, MPV/PLT, AISI, and SIRI. Serum samples were assayed for Arg, ADMA, SDMA, and MDA. Monocytes, MLR, SIRI, and MPV/PLT were higher in men, while PLT count was higher in women. COC use increased lymphocytes and lowered PLR and MLR. Smoking reduced sexually divergent parameters, especially in COC users: smoking and non-smoking COC-free women displayed six divergent parameters, while COC users displayed only two (monocytes and MPV). In addition, COC affected endothelial function, reducing ADMA and Arg. Moreover, COC-free women had lower Arg levels than men. In conclusion, COC use strongly influence the effects of tobacco smoking, which are sex and parameter specific. Further, these data stress that COC use and smoking attitude select different cohorts indicating that sex and gender studies need intersectionality.

The characterization of urinary metabolome, which provides a fingerprint for each individual, is an important step to reach personalized medicine. It is influenced by exogenous and endogenous factors; among them, we investigated sex influences on 72 organic acids measured through GC-MS analysis in the urine of 291 children (152 males; 139 females) aging 1-36 months and stratified in four groups of age. Among the 72 urinary metabolites, in all age groups, 4-hydroxy-butirate and homogentisate are found only in males, whereas 3-hydroxy-dodecanoate, methylcitrate, and phenylacetate are found only in females. Sex differences are still present after age stratification being more numerous during the first 6 months of life. The most relevant sex differences involve the mitochondria homeostasis. In females, citrate cycle, glyoxylate and dicarboxylate metabolism, alanine, aspartate, glutamate, and butanoate metabolism had the highest impact. In males, urinary organic acids were involved in phenylalanine metabolism, citrate cycle, alanine, aspartate and glutamate metabolism, butanoate metabolism, and glyoxylate and dicarboxylate metabolism. In addition, age specifically affected metabolic pathways, the phenylalanine metabolism pathway being affected by age only in males. Relevantly, the age-influenced ranking of metabolic pathways varied in the two sexes. In conclusion, sex deeply influences both quantitatively and qualitatively urinary organic acids levels, the effect of sex being age dependent. Importantly, the sex effects depend on the single organic acid; thus, in some cases the urinary organic acid reference values should be stratified according the sex and age.

OBJECTIVES This case control study sought to assess the presence and characteristics of cardiac abnormalities in patients with Alzheimer disease (AD). BACKGROUND Protein misfolding is involved in the pathophysiology of neurodegenerative disorders such as AD. Recently, amyloid-beta (Ab) aggregates were identified within the cardiomyocytes and interstitium of patients with AD, suggesting that Ab oligomers may reach and damage the heart. METHODS The authors studied 32 patients with AD and 34 controls matched by age and sex, all of whom were free from cardiac or systemic diseases. A clinical evaluation, an electrocardiogram, and an echocardiogram were performed in all subjects. Furthermore, patients with AD underwent genetic analyses (of the PSEN1, PSEN2, APP, and APOE genes). RESULTS Compared to the control group, patients with AD had a higher prevalence of low-voltage electrocardiographic QRS complexes (28% vs. 3%, respectively; p = 0.004), a lower voltage/mass ratio (p = 0.05), a greater echocardiographic interventricular septum (10.1 +/- 1.3 mm vs. 9.3 +/- 1.1 mm, respectively; p = 0.01), a greater maximum wall thickness (10.8 +/- 1.7 mm vs. 9.3 +/- 1.1 mm, respectively; p = 0.0001), and a 2-fold higher prevalence of diastolic dysfunction (70% vs. 35%, respectively; p = 0.007). Symptoms and signs of heart failure were absent in all patients with AD. CONCLUSIONS This study shows that electrocardiographic and echocardiographic abnormalities, including diastolic dysfunction, are present in patients with AD and that these studies reproduce the pattern of cardiac amyloidosis. These findings suggest that, in AD, there may be subclinical cardiac involvement likely associated with Ab amyloid deposition. The clinical relevance of these cardiac abnormalities should be evaluated in larger prospective studies. (c) 2019 by the American College of Cardiology Foundation.

Children, especially those aged <5 years, and adolescents are at increased risk of progression to active TB disease when infected. Management of childhood TB outbreaks is crucial for TB elimination especially in low burden countries. We searched the electronic databases MEDLINE-CINHAL-EMBASE up to July 2017 for primary studies reporting on TB incidents which involved teacher/child-caregiver, relative or students diagnosed with TB in a school/childcare setting or in other congregate settings attended by children and adolescents. Out of 10,481 citations, 74 studies, published mostly in low TB burden countries from 1950 to 2017, describing 128 incident investigations, were included. Overall 5025 (14.2%) LTBI and 811 (2.3%) TB cases were diagnosed among 35,331 screened individuals. Incidents occurred mainly in schools (89.1%) where index cases were more frequently students (63.3%) than teachers/caregivers; almost all of the incidents exposing children aged 2-5 were attributable to a teacher/caregiver index case. In 68 individual contact investigations the pooled proportions of TB and LTBI among those exposed were 0.03 (95% CI 0.02-0.04) and 0.15 (95% CI 0.13-0.18). The overall risk of developing TB disease in school-congregate settings seems slightly lower than in high-income country household settings. Public health interventions targeting school-congregate settings may be critical to overall TB control and towards TB elimination in low-burden countries.

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Background: Articular cartilage lacks a regenerative response. Embryonic stem cells (ESCs) are a source of pluripotent cells for cartilage regeneration. Their use, however, is associated with a risk of teratoma development, which depends on multiple factors including the number of engrafted cells and their degree of histocompatibility with recipients, the immunosuppression of the host and the site of transplantation. Colonies of sheep embryonic stem-like (ES-like) cells from in vitro-produced embryos, positive for stage-specific embryonic antigens (SSEAs), alkaline phosphatase (ALP), Oct 4, Nanog, Sox 2 and Stat 3 gene expression, and forming embryoid bodies, were pooled in groups of two-three, embedded in fibrin glue and engrafted into osteochondral defects in the left medial femoral condyles of 3 allogeneic ewes (ES). Empty defects (ED) and defects filled with cell-free glue (G) in the condyles of the controlateral stifle joint served as controls. After euthanasia at 4 years post-engraftment, the regenerated tissue was evaluated by macroscopic, histological and immunohistochemical (collagen type II) examinations and fluorescent in situ hybridization (FISH) assay to prove the ES-like cells origin of the regenerated tissue. Results: No teratoma occurred in any of the ES samples. No statistically significant macroscopic or histological differences were observed among the 3 treatment groups. FISH was positive in all the 3 ES samples. Conclusions: This in vivo preclinical study allowed a long-term evaluation of the occurrence of teratoma in non-immunosuppressed allogeneic adult sheep engrafted with allogeneic ES-like cells, supporting the safe and reliable application of ES cells in the clinic.

Haemoptysis etiologies vary widely among studies (Mondoni M et al., Eur Respir J 2016; 47:348-50). Bronchoscopy plays a key role in localizing the bleeding source (Sakr L, et al., Respiration2010; 80:38-58). We conducted a prospective, multicentre, observational study investigating hemoptysis etiology in association with the severity of the symptom. We also studied the ability of bronchoscopy to detect the bleeding source (i.e., lung and lobe) in association with the time of endoscopic examination. From July 2013 to September 2015, 475 patients, whose mean (SD) age was 64.6 (16.7) years, were enrolled in five Italian lung diseases hospital units. Pneumonia (20.4%), lung cancer (17.1%), and bronchiectasis (14.7%) were the most frequent causes of haemoptysis. In contrast to recent findings (Abdulmalak C, et al., Eur Respir J, 46:503-11) idiopatic haemoptysis accounted only for 8.6% of all causes. Lung cancer and pneumonia were the most frequent causes of moderate-severe and mild bleeding, respectively. When performed within 48 hours from the last haemoptysis episode, bronchoscopy showed a higher detection ability than after 48 hours (lung: 69/130 (53.1%) vs. 74/213 (34.7%), P: 0.001; lobe: 55/131 (42.0%) vs. 59/215 (27.4%), P: 0.005). However, no differences were found when performed within 24 vs. within 48 hours (lung: 33/58 (56.9%) vs. 69/130 (53.1%), P: 0.63; lobe: 28/59 (47.5%) vs. 55/131 (42.0), P: 0.48). Pneumonia is the leading cause of haemoptysis in our Italian cohort, while lung cancer is the most frequent cause of severe bleeding. When performed within 48 hours from the last haemoptysis episode, bronchoscopy shows a better ability to detect the bleeding source (both the lung and the lobe).

PURPOSE:To evaluate the efficacy and safety of a single intravitreal dexamethasone implant (DXI) combined with intravitreal antivascular endothelial growth factor (anti-VEGF) therapy, in patients with neovascular age-related macular degeneration (wet-AMD) resistant to conventional treatment.METHODS:In this randomized, controlled pilot study, 16 eyes of 15 patients, unresponsive to anti-VEGF therapy, were enrolled and randomly assigned to two groups: DXI + anti-VEGF (treatment group: 11 eyes) and monthly anti-VEGF alone (control group: 5 eyes). Patients were treated at baseline and followed for 6 months. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) parameters, and fluorescein angiography (FA) were evaluated.RESULTS:Eight eyes (72.7%) in the treatment group and 2 eyes in the control group (40%) showed complete retinal fluid resorption (p=0.049). BCVA showed no significant change from baseline in both the treatment group and the control group (p=0.40 and p=0.29, respectively). Both median central foveal thickness (CFT) and median macular volume showed a greater reduction from baseline in the treatment group.CONCLUSION:In patients showing an incomplete response to anti-VEGF therapy, DXI combined with intravitreal anti-VEGF seems to improve retinal fluid resorption without functional advantage. This trial is registered with ACTRN12618001102268.

Mastitis is the main cause of antibiotic use in dairy sheep. Post-milking teat disinfection is adopted to prevent intra-mammary infections. Use of chemical disinfectants is associated with biocide and antibiotic resistance. Terpinen-4-ol (T-4-ol), component of the Melaleuca alternifolia essential oil, shows antibacterial properties without inducing resistance. Aim of the trial was to evaluate the efficacy of a T-4-ol post-dipping compound in preventing bacterial mastitis in dairy sheep. In two different farms, two groups of 35 ewes were recruited as experimental and control arm (exposed to chlorhexidine-based disinfectant). No differences were observed during the follow-up. In one livestock, during the first year, 30-day incidence of mastitis was significantly lower in the T-4-ol group. Post-milking teat disinfection with T-4-ol showed equivalent efficacy in terms of mastitis incidence. Disinfectants based on natural compounds for mastitis prevention could potentially reduce environmental pollution, avoid drug residues in milk and decrease the antimicrobial resistance development.

OBJECTIVE To determine the minimum alveolar concentration of desflurane (MACDES) and effects on cardiovascular variables in positive-pressure ventilated sheep. ANIMALS 13 adult female sheep. PROCEDURES Anesthesia was induced with desflurane. After a 30-minute equilibration at an end-tidal concentration of desflurane (etDES) of 10.5%, an electrical stimulus (5 Hz/ms and 50 mA) was applied for 1 minute or until gross purposeful movement occurred. The etDES was then changed by 0.5% (modified up-down method), depending on whether a positive motor response had been elicited, and stimulation was repeated. The MACDES was the etDES midway between a positive and negative response. After MACDES was determined, etDES was increased to 1.3 and 1.6 MACDES. Animals were allowed to equilibrate for 15 minutes, and cardiovascular, blood gas, acid-base, and hematologic variables were measured. Times to induction of anesthesia, extubation, attainment of sternal position, and standing and duration of anesthesia were recorded. RESULTS Mean ± SD MACDES was 9.81 ± 0.79%. Times to intubation, extubation, and standing were 4.81 ± 2.21 minutes, 14.09 ± 4.05 minutes, and 32.4 ± 12.5 minutes, respectively. Duration of anesthesia was 226 ± 22 minutes. Heart rate increased significantly at induction of anesthesia but otherwise remained at preanesthetic rates. Arterial blood pressures progressively decreased with increasing etDES; pressures increased slightly only in response to noxious stimulation. CONCLUSIONS AND CLINICAL RELEVANCE The MACDES determined here compared favorably with that determined for other sheep populations and indicated similar anesthetic potency as in other species. Desflurane caused dose-dependent arterial hypotension, which indicated the need for careful blood pressure monitoring.

Pulmonary and extra-pulmonary diseases caused by nontuberculous mycobacteria: new clinical and public health threats http://ow.ly/87Dm30eMFd9.

Cathelicidins are well-characterized antimicrobial peptides (AMPs) that are present in significant amounts in mastitic milk. Neutrophils are believed to be the main producers of these AMPs, while the role of mammary epithelial cells (MECs) in their production and release is still unclear. In this work, cathelicidin production patterns were investigated in mammary tissues of ewes infected by Staphylococcus aureus, Streptococcus uberis, or Mycoplasma agalactiae, with a combined approach including immunohistochemistry, immune-colocalization, and fluorescent in situ hybridization. Our results confirm that MECs produce and release cathelicidins in response to different mastitis pathogens. As opposed to neutrophils, however, MECs do not seem to store the preformed protein precursor in their cytoplasm, but appear to synthesize and release it only upon exposure to the microorganisms. Cathelicidin production by MECs appears to occur before leukocyte influx in the milk, suggesting a role for these cells in the initial response of the mammary epithelium to microbial infection. Once in the milk, infiltrating neutrophils release massive amounts of cathelicidin by degranulation and production of neutrophil extracellular traps, acting as the main contributor for cathelicidin abundance in mastitic milk. Taken together, our results support the active contribution of MECs to cathelicidin production and release, and reinforce the value of cathelicidins as sensitive and pathogen-independent mastitis markers.