Bronchopneumonia with interstitial pneumonia in feedlot cattle: Epidemiologic characteristics of affected animals.

Bronchopneumonia with interstitial pneumonia (BIP) of feedlot cattle is characterized by gross and histologic lesions of cranioventral bronchopneumonia (BP) and caudodorsal interstitial pneumonia. This study described the characteristics and frequency of BIP in western Canadian feedlot cattle and identified epidemiologic differences between BIP and either BP or acute interstitial pneumonia (AIP). The study of 9909 deaths on 4 western Canadian feedlots included 1105 BIP, 1729 BP, and 878 AIP cases. A population of 55 cases with gross, histopathology, and microbiology data was used to validate the primary data set. BIP was the second most common reason for death (or euthanasia) from respiratory disease (1105/9909 cases), and the observed frequency was twice what was expected from random concurrence of BP and AIP. Based on logistic regression models, epidemiologic characteristics of BIP were comparable to those of BP, although BIP cases were more chronic with more instances of clinical illness prior to death. BIP was epidemiologically distinct from AIP. Specifically, BIP more frequently affected steers than heifers, deaths occurred earlier in the feeding period at lower body weights and lower daily weight gains, and BIP cases had longer durations from the first clinical illness to death and more separate instances of clinical illness prior to death. Furthermore, death from BIP mainly occurred in winter and fall, while death from AIP was most frequent in summer. These findings define BIP as a unique condition of feedlot cattle and suggest that chronic BP may promote the development of fatal interstitial lung disease in at-risk cattle.

Bronchopneumonia with interstitial pneumonia in beef feedlot cattle: Characterization and laboratory investigation.

Bronchopneumonia with interstitial pneumonia (BIP) has been considered a variant of acute interstitial pneumonia (AIP) rather than a distinct disease. This study compared 18 BIP, 24 bronchopneumonia (BP), and 13 AIP cases in feedlot beef cattle. Grossly, BIP cases typically had cranioventral lung lesions of similar morphology and extent as BP cases, but the caudodorsal lung appeared overinflated, bulged on section, and had interlobular edema and emphysema. Gross diagnosis of BIP had 83% sensitivity and 73% specificity relative to histopathology. Histologic lesions of BIP in cranioventral areas were of chronic BP, while caudodorsal lesions included alveolar and bronchiolar damage and inflammation, interstitial hypercellularity, and multifocal hemorrhages. In BIP cases, cranioventral lung lesions were more chronic than caudodorsal lesions. Histologic scores and microbiology data were comparable in cranioventral lung of BIP versus BP cases and caudodorsal lung of BIP versus AIP cases, with differences reflecting a more chronic disease involving less virulent bacteria in BIP versus BP. Mycoplasma bovis infection was similarly frequent among groups, and a viral cause of BIP was not identified. Lesion morphology and similar blood cytokine concentrations among groups argued against sepsis as a cause of lung injury. Surfactant dysfunction was identified in BIP and BP, and was only partially the result of protein exudation. These and other findings establish BIP as a distinct condition in which chronic cranioventral BP precedes acute caudodorsal interstitial lung disease, supporting a role of chronic inflammation in heightened sensitivity to 3-methylindole or another lung toxicant.

Impact of the Covid pandemic on health care equity in NHS General Dental Practice.

INTRODUCTION: Oral health inequalities existed before Covid, but the pandemic presented an unprecedented challenge for health services. Our aim was to determine whether patient groups at risk of health care inequality due to the pandemic could be identified from NHS dental claims. METHODS: Secondary analysis of routinely collected NHS Business Services Authority data for patients treated by General Dental Practitioners in England and Wales between April 2019 and January 2022 to assess the effect of the Covid pandemic on claims for patients attending general dental practices. Data for treatment items claimed after the start of the first lockdown were compared to the pre-lockdown period. RESULTS: The proportion of claims for child fillings, child extractions and child fluoride varnish application after March 2020 were lower than equivalent proportions for adults, in both England and Wales. Similarly, there were consistently fewer claims for fillings and extractions for patients claiming pension credit guarantee credit than all pensioners in both England and Wales. CONCLUSION: The Covid pandemic may have caused health care inequality for children and patients claiming pension credit guarantee credit. This may compound the inequality in oral health for these patients.

The vascular Ca2+-sensing receptor regulates blood vessel tone and blood pressure.

The extracellular calcium-sensing receptor CaSR is expressed in blood vessels where its role is not completely understood. In this study, we tested the hypothesis that the CaSR expressed in vascular smooth muscle cells (VSMC) is directly involved in regulation of blood pressure and blood vessel tone. Mice with targeted CaSR gene ablation from vascular smooth muscle cells (VSMC) were generated by breeding exon 7 LoxP-CaSR mice with animals in which Cre recombinase is driven by a SM22α promoter (SM22α-Cre). Wire myography performed on Cre-negative [wild-type (WT)] and Cre-positive (SM22α)CaSR(Δflox/Δflox) [knockout (KO)] mice showed an endothelium-independent reduction in aorta and mesenteric artery contractility of KO compared with WT mice in response to KCl and to phenylephrine. Increasing extracellular calcium ion (Ca(2+)) concentrations (1-5 mM) evoked contraction in WT but only relaxation in KO aortas. Accordingly, diastolic and mean arterial blood pressures of KO animals were significantly reduced compared with WT, as measured by both tail cuff and radiotelemetry. This hypotension was mostly pronounced during the animals' active phase and was not rescued by either nitric oxide-synthase inhibition with nitro-l-arginine methyl ester or by a high-salt-supplemented diet. KO animals also exhibited cardiac remodeling, bradycardia, and reduced spontaneous activity in isolated hearts and cardiomyocyte-like cells. Our findings demonstrate a role for CaSR in the cardiovascular system and suggest that physiologically relevant changes in extracellular Ca(2+) concentrations could contribute to setting blood vessel tone levels and heart rate by directly acting on the cardiovascular CaSR.

Vasopressin regulation of sodium transport in the distal nephron and collecting duct.

Arginine vasopressin (AVP) is released from the posterior pituitary gland during states of hyperosmolality or hypovolemia. AVP is a peptide hormone, with antidiuretic and antinatriuretic properties. It allows the kidneys to increase body water retention predominantly by increasing the cell surface expression of aquaporin water channels in the collecting duct alongside increasing the osmotic driving forces for water reabsorption. The antinatriuretic effects of AVP are mediated by the regulation of sodium transport throughout the distal nephron, from the thick ascending limb through to the collecting duct, which in turn partially facilitates osmotic movement of water. In this review, we will discuss the regulatory role of AVP in sodium transport and summarize the effects of AVP on various molecular targets, including the sodium-potassium-chloride cotransporter NKCC2, the thiazide-sensitive sodium-chloride cotransporter NCC, and the epithelial sodium channel ENaC.

Anaesthetic records: an evaluation of variation across England.

We collected blank non-specialist anaesthetic records from 71 National Health Service Trusts in England. A data set was established by collating all data items found in an initial tranche of 28 records. All 71 records were subsequently analysed for each data item in this data set. We found significant variation: the most populated record included 216 data items and the least included 38 data items: a greater than five-fold variation. There was significant variation in the inclusion of data items commonly considered important to patient safety; 42% of records omitted documentation of fasting status, 72% omitted documentation of a discussion around the risk of accidental awareness during general anaesthesia, 92% omitted documentation of quantitative neuromuscular blockade monitoring and 63% omitted documentation for 'Stop Before You Block' when performing regional anaesthesia. The study highlights significant variability in the composition of anaesthetic records across England which may impact on its value as a data repository, an action trigger, a medicolegal account, and a tool to facilitate safe handover. Standardisation of the anaesthetic record or the establishment of standards of recording would help to allay potential risks to patient safety and assist in guiding future procurement of electronic solutions for anaesthetic records.

Characterization of a novel phosphorylation site in the sodium-chloride cotransporter, NCC.

The sodium-chloride cotransporter, NCC, is essential for renal electrolyte balance. NCC function can be modulated by protein phosphorylation. In this study, we characterized the role and physiological regulation of a novel phosphorylation site in NCC at Ser124 (S124). Novel phospho-specific antibodies targeting pS124-NCC demonstrated a band of 160 kDa in the kidney cortex, but not medulla, which was preabsorbed by a corresponding phosphorylated peptide. Confocal microscopy with kidney tubule segment-specific markers localized pS124-NCC to all distal convoluted tubule cells. Double immunogold electron microscopy demonstrated that pS124-NCC co-localized with total NCC in the apical plasma membrane of distal convoluted tubule cells and intracellular vesicles. Acute treatment of Munich-Wistar rats or vasopressin-deficient Brattleboro rats with the vasopressin type 2 receptor-specific agonist dDAVP significantly increased pS124-NCC abundance, with no changes in total NCC plasma membrane abundance. pS124-NCC levels also increased in abundance in rats after stimulation of the renin-angiotensin-aldosterone system by dietary low sodium intake. In contrast to other NCC phosphorylation sites, the STE20/SPS1-related proline-alanine-rich kinase and oxidative stress-response kinases (SPAK and OSR1) were not able to phosphorylate NCC at S124. Protein kinase arrays identified multiple kinases that were able to bind to the region surrounding S124. Four of these kinases (IRAK2, CDK6/Cyclin D1, NLK and mTOR/FRAP) showed weak but significant phosphorylation activity at S124. In oocytes, (36)Cl uptake studies combined with biochemical analysis showed decreased activity of plasma membrane-associated NCC when replacing S124 with alanine (A) or aspartic acid (D). In novel tetracycline-inducible MDCKII-NCC cell lines, S124A and S124D mutants were able to traffic to the plasma membrane similarly to wildtype NCC.

Acute interstitial pneumonia and the biology of 3-methylindole in feedlot cattle.

Acute interstitial pneumonia (AIP) of cattle has been recognized for many decades. While the pathogenesis and risk factors for this condition in pastured cattle are relatively well characterized, there remains a poor understanding of the disease as it occurs in intensively fed cattle such as in beef feedlots. Specifically, in pastured cattle, AIP results from excessive ruminal production of the pneumotoxicant 3-methylindole (3-MI). In feedlot cattle, the evidence to substantiate the role of 3-MI is comparatively deficient and further investigations into the cause, pathogenesis, and control are sorely needed. This review highlights our current understanding of AIP with a focus on the disease as it occurs in feedlot cattle. Additionally, it illustrates the need for further work in understanding the specific animal factors (e.g. the ruminal microbiome, and the role of concurrent diseases), management factors (e.g. animal stocking and vaccination protocols), and dietary factors (e.g. dietary supplements) that may impact the development of AIP and which are relatively unique to the feedlot setting. All stakeholders in the beef industry stand to benefit from a greater understanding of what remains a pressing yet poorly understood issue in beef production.

Short communication: Effects of dietary nitrogen concentration on messenger RNA expression and protein abundance of urea transporter-B and aquaporins in ruminal papillae from lactating Holstein cows.

To test the hypothesis that dietary N concentrations affect gut epithelial urea transport by modifying the expression of urea transporter B (UT-B) and aquaporins (AQP), the mRNA expression and protein abundance of UT-B and AQP3, AQP7, AQP8, and AQP10 were investigated in ruminal papillae from 9 lactating dairy cows. Ruminal papillae were harvested from cows fed low N (12.9% crude protein) and high N (17.1% crude protein) diets in a crossover design with 21-d periods. The mRNA expression was determined by real-time reverse transcription-PCR and protein abundance by immunoblotting. The mRNA expression of UT-B was not affected by dietary treatment, whereas mRNA expression of AQP3, 7, and 10 were greater in the high N compared with the low N fed cows. Using peptide-derived rabbit antibodies to cow AQP3, 7, and 8, immunoblotting revealed bands of approximately 27, 27, and 24 kDa in ruminal papillae, respectively. A peptide-derived chicken antibody to cow UT-B detected a band of approximately 30 to 32 kDa in ruminal papillae. The abundance of UT-B and AQP3 and 7 were not affected by dietary treatment. In contrast, the abundance of AQP8 was greater in high N compared with low N diets. In conclusion, AQP3, 7, and 8 were found to be expressed in bovine rumen papillae. None of the investigated transcripts or proteins correlated to the increased rumen epithelial urea permeability observed with low dietary N concentration.

A plate reader-based method for cell water permeability measurement.

Cell volume and water permeability measurements in cultured mammalian cells are typically conducted under a light microscope. Many of the employed approaches are time consuming and not applicable to a study of confluent epithelial cell monolayers. We present here an adaptation of a calcein-quenching-based approach for a plate reader. A standard curve of fluorescence intensities at equilibrium has been recorded, following a shift from 285 mosmol/kgH(2)O to a series of altered extracellular osmolyte concentrations, ranging from final concentrations of 185 to 585 mosmol/kgH(2)O, by changing buffer d-mannitol concentrations. Similarly, according average cell volumes have been measured in suspension in a Coulter counter (particle-sizing device). Based on these measurements, we have derived an equation that facilitates the modeling of cell volume changes based on fluorescence intensity changes. We have utilized the method to study the role of a carboxyl-terminus aquaporin (AQP)-2 phosphorylation site, which is known to affect AQP2 membrane trafficking, in heterologous type I Madin-Darby canine kidney cells. We find that water permeability in cells expressing phosphorylation site mutants was in the following order: AQP2-S256D > AQP2 wild-type > AQP2-S256A. We propose that the method can be applied to study AQP function and more generally to study cell volume changes in adherent cell lines. Furthermore, it should be adaptable for AQP inhibitor screening in chemical compound libraries.

Regulation of intracellular actin polymerization by prenylated cellular proteins.

Posttranslational modification by covalent attachment of polyisoprene intermediates to a carboxyterminal CAAX-box motif is required for the biologic function of proteins such as p21ras, the supergene family of ras-related proteins, nuclear lamins, and subunits of heterotrimeric G-proteins. Cells grown in the presence of lovastatin, which inhibits HMG-CoA reductase and prevents synthesis of intermediates required for protein prenylation, develop a round, refractile morphology. Our data indicate that this is due to the selective loss of actin cables without gross changes in the microtubular lattice or intermediate filament structure. Microinjection of a competitive peptide inhibitor of protein prenyltransferases into the cytoplasm of cells induces an identical change in morphology with loss of actin cables. Mevalonate (MVA) reverses the lovastatin-induced morphologic change by inducing a rapid repolymerization of actin cables with coincident reversion to the flat morphology. Furthermore, microinjection of farnesyl-pyrophosphate or geranylgeranyl-pyrophosphate into lovastatin-treated cells also results in rapid morphologic reversion. The morphologic reversion induced by MVA requires the presence of serum, and is independent of extracellular calcium. The addition of cycloheximide to the growth medium prevents lovastatin-induced loss of actin cables, and causes morphologic reversion of lovastatin-treated cells by a mechanism that is independent of MVA. A1F4- induces morphologic reversion in a manner indistinguishable from MVA. These data indicate that prenylated protein(s) play a critical role in regulating the state of intracellular actin, and that GGPP can rescue the lovastatin-induced morphologic phenotype in the absence of upstream intermediates of cholesterol biosynthesis. We have begun to dissect the signaling events that mediate this pathway.

Isotypic exclusion of gamma delta T cell receptors in transgenic mice bearing a rearranged beta-chain gene.

The rearrangement of T cell antigen receptor beta- and gamma-chain gene segments was studied in transgenic mice that bear a functional beta-chain gene. Virtually all CD3-positive T cells derived from transgenic mice express beta chains containing the transgene-encoded V beta 8.2 variable region on their surfaces and do not express endogenous beta-chain variable regions. Expression of endogenous V beta genes is inhibited at the level of somatic recombination during thymic ontogeny. Furthermore, rearrangements of the TCR gamma-chain genes are also markedly inhibited in these transgenic animals. Hence expression of the TCR beta transgene has led to allelic exclusion of alpha beta receptors and isotypic exclusion of gamma delta T cell receptors.

Efficacy of a florfenicol-flunixin meglumine combination product versus tulathromycin or ceftiofur crystalline free acid for the treatment of undifferentiated fever in feedlot calves.

In this field trial, a new combination product containing florfenicol and flunixin meglumine (FLOR-FM) was compared with commercially available products that contained only tulathromycin (TULA) or ceftiofur crystalline free acid (CCFA) for the treatment of undifferentiated fever (UF; rectal temperature >/=105.0 degrees F) in beef calves that received long-acting oxytetracycline at feedlot arrival. The overall mortality rate of the FLOR FM group (2.0%) was significantly (P less than .050) lower than the rates in the TULA and CCFA groups (10.0% and 20.0%, respectively; 50 animals/group), even though the first UF relapse rate of the FLOR FM group was significantly (P less than .050) higher than that of the TULA group. In the FLOR FM group, this resulted in per-animal economic advantages of Can$46.23 (versus TULA) and Can$108.77 (versus CCFA) based on equal costs for initial UF therapy. These results demonstrate that it is more cost-effective to administer FLOR FM than TULA or CCFA for initial UF therapy in feedlot calves at high risk for bovine respiratory disease that receive metaphylactic long-acting oxytetracycline at feedlot arrival.

What is your diagnosis? Perifemoral mass in a cow.

A 15-year-old female Simmental cross-breed cow was presented to the Purdue University Veterinary Teaching Hospital for evaluation of a perifemoral soft tissue mass. Impression smears made from an excisional biopsy contained a population of pleomorphic mesenchymal cells with abundant, periodic acid-Schiff-positive (PAS), intracytoplasmic granular material, and rare elongated multinucleated cells consistent with strap-like cells. A second population of small round cells suggestive of lymphocytes or progenitor cells was also noted. A cytologic diagnosis of sarcoma was made, with rhabdomyosarcoma considered most likely based on the large amount of PAS-positive material (presumed to be glycogen) and the rare strap-like cells. Histopathologic sections contained an unencapsulated, densely cellular neoplasm composed of haphazardly arranged highly pleomorphic mesenchymal cells and a few small round cells. The mesenchymal cells were positive for vimentin, non-specific muscle actin, and myoglobin, and negative for phosphotungstic acid-hematoxylin, smooth muscle actin, and desmin. Glycogen granules were confirmed by transmission electron microscopy. A diagnosis of pleomorphic rhabdomyosarcoma was made. While cytologic findings may suggest rhabdomyosarcoma, cytologic features can be highly variable, and a definitive diagnosis usually requires cytochemical and immunohistochemical staining.

Spontaneous middle-ear encephalocele: report of two cases and brief review.

We report two cases of middle-ear spontaneous (idiopathic) encephalocele in patients aged 69 and 82 years. To our knowledge the latter represents the oldest individual reported to have such a lesion. Microscopic examination shows a disorganized neuropil with reactive inflammatory changes and in one case, several cysts lined by simple cylindrical ciliated epithelium. Encephalocele must be included in the differential diagnoses of otorrhea and masses in the middle ear.

Evaluation of the efficacy and cost-effectiveness of melengestrol acetate in feedlot heifer calves in western Canada.

The purpose of this study was to evaluate the relative efficacy and cost-effectiveness of feeding melengestrol acetate (MGA) to feedlot heifer calves in western Canada. Heifers fed MGA had significantly (P less than .05) improved average daily gain, feed conversion, and carcass quality grade and lower rates of initial undifferentiated fever treatment and bovine respiratory disease mortality. However, heifers fed MGA had less desirable (P less than .05) carcass yield grade. There was a net economic advantage of Can $11.31/animal in favor of heifers fed MGA. Based on these results, it is efficacious and cost-effective to feed MGA to heifer calves raised in standard large-pen commercial feedlots in western Canada.

A comparison of florfenicol and tulathromycine for the treatment of undifferentiated fever in feedlot calves.

The purpose of this study was to compare the relative cost-effectiveness of florfenicol with that of tulathromycin for treatment of undifferentiated fever (UF) in feedlot calves at ultra-high risk of developing UF that receive metaphylactic tulathromycin on arrival at the feedlot. Calves that received therapeutic florfenicol had lower overall mortality (P=.045) and bovine respiratory disease mortality (P=.050) compared with calves that received therapeutic tulathromycin, but no significant differences were detected in feedlot performance, carcass characteristics, or other animal health variables. There was a net advantage of Can$41.19/treated animal in the florfenicol group versus the tulathromycin group. This study demonstrates that it is more cost-effective to use florfenicol than tulathromycin for the initial treatment of UF in feedlot calves at ultra-high risk of developing UF that receive on-arrival metaphylactic tulathromycin.

A comparison of 2 vaccination programs in feedlot calves at ultra-high risk of developing undifferentiated fever/bovine respiratory disease.

The aim of this study was to compare 2 vaccination programs in feedlot calves at ultra-high risk of developing undifferentiated fever (UF)/bovine respiratory disease (BRD). At feedlot arrival, 3882 calves were enrolled in the study and randomly allocated to 2 groups, which were housed by group in 12 pens. At the time of allocation, 1 group (MLV3-BT2) received a multivalent, modified-live viral vaccine containing infectious bovine rhinotracheitis virus (IBRV) and types I and II bovine viral diarrhea virus (BVDV), as well as a Mannheimia haemolytica (MH) and Pasteurella multocida bacterin-toxoid. The other group (MLV4-BT1) received a vaccine containing IIBRV, type I BVDV, bovine respiratory syncytial virus, and parainfluenza-3 virus, as well as a MH bacterin-toxoid. At an average of 69 days post arrival, the groups received their respective viral vaccines. The initial UF treatment, overall chronicity, overall wastage, overall mortality, and BRD mortality rates were significantly (P < 0.05) lower in the MLV3-BT2 group than in the MLV4-BT1 group. Average daily gain and the proportions of yield grade Canada 3 and quality grade E carcasses were significantly (P < 0.05) higher in the MLV3-BT2 group than in the MLV4-BT1 group. No significant (P > or = 0.05) difference in the dry matter intake to gain ratio was detected between the 2 groups. In economic terms, there was a net advantage of $20.86 CDN/animal in the MLV3-BT2 group. This study demonstrates that it is more cost effective to use an MLV3-BT2 vaccination program than a MLV4-BT1 vaccination program in feedlot calves at ultra-high risk of developing UF/BRD.

Microbiological and histopathological findings in cases of fatal bovine respiratory disease of feedlot cattle in Western Canada.

The aim of this study was to describe the microbiologic agents and pathologic processes in fatal bovine respiratory disease (BRD) of feedlot cattle and to investigate associations between agents and pathologic processes. Ninety feedlot calves diagnosed at necropsy with BRD and 9 control calves without BRD were examined, using immunohistochemical (IHC) staining and histopathologic studies. Mannheimia haemolytica (MH) (peracute, acute, and subacute cases) and Mycoplasma bovis (MB) (subacute, bronchiolar, and chronic cases) were the most common agents identified in fatal BRD cases. Significant associations (P < 0.10) were detected between microbiologic agents and between agents and pathologic processes. When IHC staining was used, 25/26 (96%) of animals that were positive for bovine viral diarrhea virus (BVDV) were also positive for MH; 12/15 (80 %) of animals that were positive for Histophilus somni (HS) were also positive for MB; and all of the animals that were positive for HS were negative for MH and BVDV. This quantitative pathological study demonstrates that several etiologic agents and pathologic processes are involved in fatal BRD of feedlot cattle.

Cell transformation mediated by chromosomal deoxyribonucleic acid of polyoma virus-transformed cells.

To study the mechanism of deoxyribonucleic acid (DNA)-mediated gene transfer, normal rat cells were transfected with total cellular DNA extracted from polyoma virus-transformed cells. This resulted in the appearance of the transformed phenotype in 1 X 10(-6) to 3 X 10(-6) of the transfected cells. Transformation was invariably associated with the acquisition of integrated viral DNA sequences characteristic of the donor DNA. This was caused not by the integration of free DNA molecules, but by the transfer of large DNA fragments (10 to 20 kilobases) containing linked cellular and viral sequences. Although Southern blot analysis showed that integration did not appear to occur in a homologous region of the recipient chromosome, the frequency of transformation was rather high when compared with that of purified polyoma DNA, perhaps due to "position" effects or to the high efficiency of recombination of large DNA fragments.