SR-B1 drives endothelial cell LDL transcytosis via DOCK4 to promote atherosclerosis.

Atherosclerosis, which underlies life-threatening cardiovascular disorders such as myocardial infarction and stroke1, is initiated by passage of low-density lipoprotein (LDL) cholesterol into the artery wall and its engulfment by macrophages, which leads to foam cell formation and lesion development2,3. It is unclear how circulating LDL enters the artery wall to instigate atherosclerosis. Here we show in mice that scavenger receptor class B type 1 (SR-B1) in endothelial cells mediates the delivery of LDL into arteries and its accumulation by artery wall macrophages, thereby promoting atherosclerosis. LDL particles are colocalized with SR-B1 in endothelial cell intracellular vesicles in vivo, and transcytosis of LDL across endothelial monolayers requires its direct binding to SR-B1 and an eight-amino-acid cytoplasmic domain of the receptor that recruits the guanine nucleotide exchange factor dedicator of cytokinesis 4 (DOCK4)4. DOCK4 promotes internalization of SR-B1 and transport of LDL by coupling the binding of LDL to SR-B1 with activation of RAC1. The expression of SR-B1 and DOCK4 is increased in atherosclerosis-prone regions of the mouse aorta before lesion formation, and in human atherosclerotic arteries when compared with normal arteries. These findings challenge the long-held concept that atherogenesis involves passive movement of LDL across a compromised endothelial barrier. Interventions that inhibit the endothelial delivery of LDL into artery walls may represent a new therapeutic category in the battle against cardiovascular disease.

Acquired Perforating Osteoma Cutis: A Rare Histopathological Diagnosis.

ABSTRACT: Perforating osteoma cutis is a benign proliferation of mature bone within the dermis and subcutaneous tissue of the skin with transepidermal elimination. Transepidermal elimination of bone is the hallmark of perforating osteoma cutis and is defined by the breaching of bone through the epidermis. Perforating osteoma cutis is exceptionally rare because only 6 cases have been recorded in the literature at the time of preparation of this report. In this report, we present the case of a 65-year-old female patient with a medical history of nonmelanoma skin cancer, hypertension, hyperlipidemia, and type II diabetes mellitus presented for evaluation of a skin lesion of the posterior lower left leg, which had been present for 1 year. Clinical and histopathologic findings were consistent with the diagnosis of acquired perforating osteoma cutis. Treatment with surgical removal by tangential biopsy has thus far proven to be both diagnostic and therapeutic because no recurrence has been noted as of 6 months.

Identifying Erythrocyte Injury in Toxicology Studies.

The hematological impacts of a drug can affect erythropoiesis at the level of the bone marrow, or decrease the life span of the RBC (red blood cell). The most common and recognizable clinical manifestation of either type of drug-induced erythropoietic injury is a decrease in RBC mass, or what is clinically referred to as an anemia. A decrease in RBC production can generally be separated from increased destruction (hemolysis) by evaluation of the hemogram for evidence of regeneration. In most healthy mammalian species, hemolysis will result in a regenerative response characterized by an increase in circulating reticulocytes. Hemorrhage as an alternative cause of a regenerative anemia can generally be excluded by careful clinical evaluation of the animal. Subsequently, the investigation of a drug-induced regenerative anemia should involve a very thorough evaluation of RBC morphology for evidence of immune-mediated destruction, RBC oxidative injury, and fragmentation that can help to identify the underlying pathological mechanism(s) involved.

Review of Histological Grading Systems in Veterinary Medicine.

Tumor grading is a method to quantify the putative clinical aggressiveness of a neoplasm based on specific histological features. A good grading system should be simple, easy to use, reproducible, and accurately segregate tumors into those with low versus high risk. The aim of this review is to summarize the histological and, when available, cytological grading systems applied in veterinary pathology, providing information regarding their prognostic impact, reproducibility, usefulness, and shortcomings. Most of the grading schemes used in veterinary medicine are developed for common tumor entities. Grading systems exist for soft tissue sarcoma, osteosarcoma, multilobular tumor of bone, mast cell tumor, lymphoma, mammary carcinoma, pulmonary carcinoma, urothelial carcinoma, renal cell carcinoma, prostatic carcinoma, and central nervous system tumors. The prognostic relevance of many grading schemes has been demonstrated, but for some tumor types the usefulness of grading remains controversial. Furthermore, validation studies are available only for a minority of the grading systems. Contrasting data on the prognostic power of some grading systems, lack of detailed instructions in the materials and methods in some studies, and lack of data on reproducibility and validation studies are discussed for the relevant grading systems. Awareness of the limitations of grading is necessary for pathologists and oncologists to use these systems appropriately and to drive initiatives for their improvement.

Sex-specific hepatic lipid and bile acid metabolism alterations in Fancd2-deficient mice following dietary challenge.

Defects in the Fanconi anemia (FA) DNA damage-response pathway result in genomic instability, developmental defects, hematopoietic failure, cancer predisposition, and metabolic disorders. The endogenous sources of damage contributing to FA phenotypes and the links between FA and metabolic disease remain poorly understood. Here, using mice lacking the Fancd2 gene, encoding a central FA pathway component, we investigated whether the FA pathway protects against metabolic challenges. Fancd2-/- and wildtype (WT) mice were fed a standard diet (SD), a diet enriched in fat, cholesterol, and cholic acid (Paigen diet), or a diet enriched in lipid alone (high-fat diet (HFD)). Fancd2-/- mice developed hepatobiliary disease and exhibited decreased survival when fed a Paigen diet but not a HFD. Male Paigen diet-fed mice lacking Fancd2 had significant biliary hyperplasia, increased serum bile acid concentration, and increased hepatic pathology. In contrast, female mice were similarly impacted by Paigen diet feeding regardless of Fancd2 status. Upon Paigen diet challenge, male Fancd2-/- mice had altered expression of genes encoding hepatic bile acid transporters and cholesterol and fatty acid metabolism proteins, including Scp2/x, Abcg5/8, Abca1, Ldlr, Srebf1, and Scd-1 Untargeted lipidomic profiling in liver tissue revealed 132 lipid species, including sphingolipids, glycerophospholipids, and glycerolipids, that differed significantly in abundance depending on Fancd2 status in male mice. We conclude that the FA pathway has sex-specific impacts on hepatic lipid and bile acid metabolism, findings that expand the known functions of the FA pathway and may provide mechanistic insight into the metabolic disease predisposition in individuals with FA.

The effect of ex vivo lipopolysaccharide stimulation and nutrient availability on transition cow innate immune cell AKT/mTOR pathway responsiveness.

Postpartum dairy cows experience a heightened inflammatory state coinciding with the time of greatest nutrient deficit. Nutrient availability is sensed on the cellular level by nutrient sensing kinases, such as the PI3K/AKT/mTOR (mTOR) pathway, a key orchestrator of immune cell activation and inflammatory balance. Our objective was to determine the responsiveness of this pathway to inflammatory stimulation with and without nutrient supplementation ex vivo. Blood samples were collected from Holstein cows (n = 14) at -42, -14, 7, 21, and 42 d relative to calving. Control samples and samples pretreated with a mixture of amino acids, glucose, and insulin (AAM) were stimulated with 100 ng/mL E. coli lipopolysaccharide (LPS; LPS, AAMLPS) or left unstimulated (control, AAM). After 1 h, ratios of mean fluorescence intensity for phosphorylated to total protein of AKT and mTORC1 substrates S6RP and 4EBP1 were analyzed in polymorphonuclear cells (PMN), and monocytes by flow cytometry. A separate aliquot was stimulated with LPS for 2 h and relative mRNA abundance of IL10, IL12A, IL12B, and TNFA in whole blood leukocytes from 10 cows was measured by reverse-transcription quantitative PCR. Repeated measures ANOVA was performed with fixed effects of time, treatment, and their interaction. Cells had different ratios of pathway proteins with PMN having the highest phosphorylation of AKT, S6RP, and 4EBP1. Stimulation with LPS consistently activated mTOR signaling in PMN regardless of nutrient supplementation except for postpartum 4EBP1, which increased in response to nutrients alone. In monocytes, AKT baseline phosphorylation was lower and activation could not be induced by either treatment, whereas activation of 4EBP1 responded to nutrient supplementation. Treatment with LPS increased phosphorylation of S6RP in both innate immune cell types. Nutrient supplementation increased baseline IL10 expression and decreased baseline as well as LPS-induced IL12B and TNFA expression. We conclude that the mTOR pathway in bovine innate immune cells can be differentially activated in response to inflammatory stimulation and nutrient supplementation in monocytes versus PMN. Effects of nutrient supplementation on cytokine mRNA abundance are likely specific to immune cell type.

C1 inhibitor in canine intravascular hemolysis (C1INCH): study protocol for a randomized controlled trial.

BACKGROUND: Immune-mediated hemolytic anemia (IMHA) is a common disease that affects all breeds of dogs and is associated with significant morbidity and mortality. Intravascular hemolysis of erythrocytes in IMHA is caused by complement activation and is often fatal. No current treatments target complement activation in canine IMHA. Human C1 esterase (C1-INH) reduces canine complement-mediated hemolysis in vitro, and a recent pharmacokinetic analysis of an FDA licensed formulation of C1-INH in dogs confirmed that a 50 IU/kg dose of C1-INH is safe to administer to dogs, and effectively inhibits canine complement mediated hemolysis ex-vivo. The C1INCH randomized controlled trial will evaluate the efficacy of this drug in dogs with intravascular hemolysis. METHODS: We will conduct a multicenter, placebo-controlled double-blind randomized clinical trial of C1-INH in dogs with intravascular hemolysis due to IMHA. We will randomize 18 dogs to receive three doses of intravenous C1-INH or saline in 24 h. Immunosuppressive and antithrombotic therapies will be standardized. Primary outcome measures will be changes in plasma free hemoglobin, serum concentrations of LDH, bilirubin, and haptoglobin. Using patient samples, we will evaluate complement activation in canine IMHA using a novel C5b-9 ELISA assay, flow cytometric detection of C3b on RBC, and by measurement of residual plasma complement activity. Secondary outcome measures will be survival to hospital discharge, duration of hospitalization, number and volume of red blood cell transfusions, and rescue therapy requirements. We will monitor dogs for adverse drug reactions. Sample size was estimated from pilot data on LDH and hemolysis index (HI) in dogs with IMHA. To detect 2-way differences between the upper and lower 50% of the LDH and HI values of equivalent size with 80% power at P < 0.05 will require 9 dogs in each arm. DISCUSSION: We anticipate that IV administration of C1-INH will significantly inhibit complement mediated hemolysis in dogs with intravascular IMHA, as determined by blood biomarker measurements (decreased plasma hemoglobin, LDH and bilirubin, increased haptoglobin). We expect this will translate into significant reductions in transfusion requirements and duration of hospitalization. TRIAL REGISTRATION: This trial has been prospectively registered with the AVMA registry (AAHSD005025).

Clinical Pathology of Donkeys and Mules.

Given the stoic nature of donkeys and their hybrids, it is important to consider the significance of diagnostic testing modalities that can provide objective health status information beyond the basic physical examination findings. However, clinical pathology assays are also fraught with significant limitations because the results for donkeys, mules, and hinnies can be difficult to interpret, and transference of data from the horse is not always applicable. This article presents considerations for sample collection, storage, analysis, and interpretation strategies for clinical pathology testing of donkeys and their hybrids based on the limited information available in the literature.

Influence of multiple reuse and resterilization cycles on the performance of a bipolar vessel sealing device (LigaSure) intended for single use.

OBJECTIVE: To determine the number of use/cleaning/resterilization cycles that can be safely applied to a vessel sealing device intended for single use (LigaSure). STUDY DESIGN: Ex vivo study. SAMPLE POPULATION: LigaSure Small Jaw handsets (n = 6) and LigaSure Impact handsets (n = 6). METHODS: Handsets underwent simulated splenectomy/cleaning/resterilization cycles until failure, defined as leaking vascular seal or blade retraction failure. Functional testing included assessment of vascular seal integrity, handset activation/tissue release, and cutting blade wear/retraction. Vascular seal failure was defined as a leak occurring at <300 mm Hg. Cycles to failure were recorded. Sealed vessels were evaluated by histology at first handset use and failure. RESULTS: Vascular seals created with the Small Jaw handset failed at a mean (95% CI) of 17.2 cycles (9.6-24.8) and a minimum of 10 cycles. Vascular seals created with the Impact failed at a mean of 20 cycles (18.4-21.6) and a minimum of 17 cycles. The majority of seal failures (73%; 95% CI 39%-94%) immediate leaked during vessel filling. The rate of vascular seal failure increased after the initial failure. Failure was associated with histologic disparities in tissue apposition. CONCLUSION: Repeated use and resterilization resulted in failure of the vascular seal due to inadequate tissue apposition after a minimum of 10 cycles. CLINICAL SIGNIFICANCE: Surgeons reusing and resterilizing LigaSure handsets (ForceTriad platform) should consider discarding handsets after 9 cycles for the Small Jaw and after 16 cycles for the Impact. Handsets should be immediately discarded after any intraoperative identification of vascular seal failure.

A Prospective Randomized Clinical Trial of a Novel, Noninvasive Perfusion Enhancement System for the Prevention of Hospital-Acquired Sacral Pressure Injuries.

PURPOSE: The purpose of this study was to determine the effectiveness of a novel, noninvasive perfusion enhancement system versus beds with integrated alternating pressure capabilities for the prevention of hospital-acquired sacral region (sacral, coccygeal, and ischium) pressure injuries in a high-risk, acute care patient population. DESIGN: A prospective randomized trial of high-risk inpatients without preexisting sacral region pressure injuries was conducted. SUBJECTS AND SETTING: The sample comprised 431 randomly enrolled adult patients in a 300-bed tertiary care community teaching hospital. METHODS: Subjects were randomly allocated to one of 2 groups: control and experimental. Both groups received "standard-of-care" pressure injury prevention measures per hospital policy, and both were placed on alternating pressure beds during their hospital stays. In addition, patients in the experimental group used a noninvasive perfusion enhancement system placed on top of their alternating pressure beds and recovery chairs throughout their hospital stay. Fischer's exact probability test was used to compare group differences, and odds ratio (OR) were calculated for comparing pressure injury rates in the experimental and control groups. RESULTS: Three hundred ninety-nine patients completed the trial; 186 patients were allocated to the experimental group and 213 patients to the control group. Eleven patients in the control group versus 2 in the experimental group developed hospital-acquired sacral region pressure injuries (51.6% vs 1.07%; P = .024). Control patients were 5.04 times more likely to develop hospital-acquired sacral region pressure injuries (OR = 0.1996; 95% CI, 0.0437-0.9125). CONCLUSIONS: Patients using a noninvasive perfusion enhancement system developed significantly fewer hospital-acquired sacral pressure injuries than those using an alternating pressure bed without the perfusion enhancement system. These findings suggest that a perfusion enhancement system enhances the success of use of pressure redistributing beds for prevention of hospital-acquired sacral pressure injuries.

Interactions of Histophilus somni with Host Cells.

Histophilus somni resides as part of the normal microflora in the upper respiratory tract of healthy cattle. From this site, the organism can make its way into the lower respiratory tract, where it is one of the important bacterial agents of the respiratory disease complex. If H. somni cells disseminate to the bloodstream, they frequently result in thrombus formation. A series of in vitro investigations have examined potential mechanisms that might contribute to such thrombus formation. Earlier work showed that H. somni can stimulate some bovine endothelial cells to undergo apoptosis. More recent studies indicate that H. somni stimulates endothelial cell tissue factor activity and disrupts intercellular junctions. The net effect is to enhance procoagulant activity on the endothelium surface and to make the endothelial monolayer more permeable to molecules, leukocytes, and perhaps H. somni cells. H. somni also activates bovine platelets, which also can enhance tissue factor activity on the endothelium surface. When exposed to H. somni, bovine neutrophils and mononuclear phagocytes form extracellular traps in vitro. Ongoing research is investigating how the interplay among endothelial cells, platelets, and leukocytes might contribute to the thrombus formation seen in infected cattle.

EVALUATION OF THE I-STAT PORTABLE CLINICAL ANALYZER FOR MEASUREMENT OF IONIZED CALCIUM AND SELECTED BLOOD CHEMISTRY VALUES IN ASIAN ELEPHANTS (ELEPHAS MAXIMUS).

Thei-STAT® portable clinical analyzer (PCA) provides patient-side results for hematologic, biochemical, and blood gas values when immediate results are desired. This analyzer is commonly used in nondomestic animals; however, validation of this method in comparison with traditional benchtop methods should be performed for each species. In this study, the i-STAT PCA was compared with the Radiometer ABL 800 Flex benchtop analyzer using 24 heparinized whole blood samples obtained from healthy E. maximus . In addition, the effect of sample storage was evaluated on the i-STAT PCA. Analytes evaluated were hydrogen ion concentration (pH), glucose, potassium (K+), sodium (Na+), bicarbonate (HCO3-), total carbon dioxide (TCO2), partial pressure of carbon dioxide (PCO2), and ionized calcium (iCa2+). Statistical analysis using correlation coefficients, Passing-Bablok regression analysis, and Bland-Altman plots found good agreement between results from samples run immediately after phlebotomy and 4 hr postsampling on the i-STAT PCA with the exception of K+, which is known to change with sample storage. Comparison of the results from the two analyzers at 4 hr postsampling found very strong or strong correlation in all values except K+, with statistically significant bias in all values except glucose and PCO2. Despite bias, mean differences assessed via Bland-Altman plots were clinically acceptable for all analytes excluding K+. Within the reference range for iCa2+, the iCa2+ values obtained by the i-STAT PCA and Radiometer ABL 800 Flex were close in value, however in light of the constant and proportionate biases detected, overestimation at higher values and underestimation at lower values of iCa2+ by the i-STAT PCA would be of potential concern. This study supports the use of the i-STAT PCA for the evaluation of these analytes, with the exception of K+, in the Asian elephant.

HEPATIC OSTEODYSTROPHY IN A GOLDEN LION TAMARIN (LEONTOPITHECUS ROSALIA).

An 8-yr-old, captive, female golden lion tamarin ( Leontopithecus rosalia ) with a 6-yr history of hyperbilirubinemia was examined for inappetence and weight loss. Physical examination and blood pressure monitoring under anesthesia revealed hypothermia and hypotension, and blood work revealed hypoglycemia, markedly elevated liver enzymes, including serum alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase, and confirmed the hyperbilirubinemia. A complete blood count suggested chronic lymphoid leukemia. The animal's condition deteriorated during recovery, and the animal died despite aggressive treatment. Grossly, there was micronodular cirrhosis of the liver, severe icterus, and diffuse osteopenia of all examined bones. Microscopic examination of the liver confirmed the micronodular cirrhosis and bone lesions were compatible with diffuse osteopenia and osteomalacia. This brief communication presents a case of chronic liver disease and lesions indicative of metabolic bone disease, also known as hepatic osteodystrophy. To the authors' knowledge, this is the first documented case of hepatic osteodystrophy in the veterinary literature.

Pregnancy-specific skin disorders.

The pregnancy-specific skin disorders are pruritic, inflammatory eruptions. The current classification by Ambros-Rudolph et al. includes four entities: pemphigoid gestationis (PG), polymorphic eruption of pregnancy (PEP), atopic eruption of pregnancy (AEP), and intrahepatic cholestasis of pregnancy (ICP). Although these disorders are all characterized by intense pruritus during pregnancy, they can be distinguished by timing, morphology, histopathology, treatment and potential for fetal complications. Diagnosis is made by clinical presentation, histology, and immunofluorescence. PEP and AEP typically resolve without sequelae; however, PG may lead to prematurity and low birth weight, and ICP is associated with an increased risk of prematurity, fetal distress, and intrauterine fetal demise. The potential for serious fetal complications necessitates a thorough evaluation of pregnancy-related pruritus. This article will discuss the skin disorders specific to pregnancy, with a focus on clinical presentation, potential for fetal complications, pathogenesis, diagnosis, and treatment.

A Novel Hazard for an Endangered Fox in a Novel Environment.

Animals colonizing novel environments can encounter novel hazards. Endangered San Joaquin kit foxes (Vulpes macrotis mutica) are found in the cities of Bakersfield and Taft in central California, USA. We documented 66 incidents of kit foxes becoming entangled in sports netting (e.g., soccer nets, batting-cage nets) occurring from the 1980s through 2022. Overall, 25 of the foxes died. Adults were more likely to get entangled in soccer nets, whereas pups (<1 yr) were more likely to get entangled in batting-cage nets. Pups are more likely to die while entangled, probably due to smaller body mass and lower energy reserves. The reasons that kit foxes get entangled in netting were unclear, although incidents involving batting-cage netting and pups may be due to natal dens being located under or near batting cages. At current rates, this hazard is unlikely to limit urban kit fox populations. However, losses of this endangered species should be minimized and the incidents are easily mitigated by dropping or lifting nets when not in use.

Clustering analysis of lipoprotein profiles to identify subtypes of hypertriglyceridemia in Miniature Schnauzers.

BACKGROUND: Hypertriglyceridemia (HTG) is prevalent in Miniature Schnauzers, predisposing them to life-threatening diseases. Varied responses to management strategies suggest the possibility of multiple subtypes. HYPOTHESIS/OBJECTIVE: To identify and characterize HTG subtypes in Miniature Schnauzers through cluster analysis of lipoprotein profiles. We hypothesize that multiple phenotypes of primary HTG exist in this breed. ANIMALS: Twenty Miniature Schnauzers with normal serum triglyceride concentration (NTG), 25 with primary HTG, and 5 with secondary HTG. METHODS: Cross-sectional study using archived samples. Lipoprotein profiles, generated using continuous lipoprotein density profiling, were clustered with hierarchical cluster analysis. Clinical data (age, sex, body condition score, and dietary fat content) was compared between clusters. RESULTS: Six clusters were identified. Dogs with primary HTG were dispersed among 4 clusters. One cluster showed the highest intensities for triglyceride-rich lipoprotein (TRL) and low-density lipoprotein (LDL) fractions and also included 4 dogs with secondary HTG. Two clusters had moderately high TRL fraction intensities and low-to-intermediate LDL intensities. The fourth cluster had high LDL but variable TRL fraction intensities with equal numbers of NTG and mild HTG dogs. The final 2 clusters comprised only NTG dogs with low TRL intensities and low-to-intermediate LDL intensities. The clusters did not appear to be driven by differences in the clinical data. CONCLUSIONS AND CLINICAL IMPORTANCE: The results of this study support a spectrum of lipoprotein phenotypes within Miniature Schnauzers that cannot be predicted by triglyceride concentration alone. Lipoprotein profiling might be useful to determine if subtypes have different origins, clinical consequences, and response to treatment.

Syntopy between Endangered San Joaquin Kit Foxes and Potential Competitors in an Urban Environment.

The endangered San Joaquin kit fox (Vulpes macrotis mutica; SJKF) occurs in the city of Bakersfield, CA, where several putative competitors also occur, including domestic cats (Felis catus), striped skunks (Mephitis mephitis), raccoons (Procyon lotor), and opossums (Didephis virginiana). We used data from a multi-year (2015-2022) city-wide camera station survey to assess whether the other species were simply sympatric with SJKF or coexisting syntopically (i.e., occurring in the same habitats without apparent competition). Annual detection rates for the other species were not correlated with SJKF rates either within SJKF habitat suitability categories (low, medium, and high) or for all categories combined. Also, detection rates for the other species did not increase in response to a significant decline in SJKF abundance caused by sarcoptic mange. The use of all SJKF habitat suitability categories by the other species and co-detections with SJKF at camera stations indicate high spatial overlap. Interference and exploitative competition between the species are apparently negligible, likely due to similar body sizes and high resource abundance. Thus, SJKF and the other species appear to be coexisting syntopically in the urban environment, resulting in a significant additional SJKF population that facilitates range-wide conservation and recovery of this endangered species.

MOVEMENTS BY SAN JOAQUIN KIT FOXES (VULPES MACROTIS MUTICA) BETWEEN URBAN AND NONURBAN HABITATS: IMPLICATIONS FOR INTERPOPULATION DISEASE TRANSFER.

Sarcoptic mange epidemics erupted in two of the remaining populations of endangered San Joaquin kit foxes (Vulpes macrotis mutica). Both populations are in urban habitats in the cities of Bakersfield and Taft, California, USA. The risk of disease spread from the two urban populations to nearby nonurban populations, and then throughout the species range, is of considerable conservation concern. To date, mange has not been detected in any nonurban populations despite considerable surveillance effort. The reasons for the lack of detections of mange among nonurban foxes are unknown. We monitored urban kit fox movements using geographic positioning system (GPS) collars to test the hypothesis that urban foxes were not venturing into nonurban habitats. Of 24 foxes monitored December 2018 to November 2019, 19 (79%) made excursions from urban into nonurban habitats from 1-124 times. The mean number of excursions per 30 d was 5.5 (range 0.1-13.9 d). The mean proportion of locations in nonurban habitats was 29.0% (range 0.6-99.7%). The mean maximum distance that foxes traveled into nonurban areas from the urban-nonurban interface was 1.1 km (range 0.1-2.9 km). Mean number of excursions, proportion of nonurban locations, and maximum distance into nonurban habitats were similar between Bakersfield and Taft, females and males, and adults and juveniles. At least eight foxes apparently used dens in nonurban habitats; shared use of dens may be an important mode of mange mite transmission between conspecifics. Two of the collared foxes died of mange during the study and two others had mange when captured at the end of the study. Three of these four foxes had made excursions into nonurban habitats. These results confirm a significant potential for mange to spread from urban to nonurban kit fox populations. We recommend continued surveillance in nonurban populations and continued treatment efforts in the affected urban populations.

Bone marrow iron scoring in healthy and clinically ill dogs with and without evidence of iron-restricted erythropoiesis.

BACKGROUND: There are few reports in dogs that have evaluated the utility of semi-quantitative scoring of bone marrow iron stores in conjunction with reticulocyte hemoglobin (CHr) to identify iron-restricted erythropoiesis due to absolute iron deficiency or iron sequestration. OBJECTIVES: An established system for scoring iron stores in human bone marrow samples was applied to dogs. The objectives were to evaluate interobserver agreement (Κω ), determine marrow iron scores in dogs without detectable hematologic abnormalities, and assess combined interpretation of iron scores and CHr to evaluate for iron-restricted erythropoiesis. METHODS: Four blinded observers independently scored iron in 139 Prussian blue-stained canine marrow samples from 0 (none) to 6 (very heavy), including healthy controls (n = 12), clinically ill dogs with (n = 100) and without (n = 16) detectable hematologic abnormalities, and dogs with experimental nutritional iron deficiency (n = 11). Additional medical record data were available for 118 dogs to evaluate for other evidence of iron deficiency (abnormal CHr, RBC indices, serum iron variables, external blood loss, or nutritional deficiencies). RESULTS: Mean Κω was 0.69 (substantial agreement) for all samples but was 0.44 (moderate agreement) for samples with iron scores <3, indicating distinguishing scores 0-2 may not be reliable. Dogs without detectable hematologic abnormalities had scores from 3-5. Dogs with scores <3 and decreased CHr often had more indicators of iron deficiency vs dogs only having low iron scores or low CHr. CONCLUSIONS: Evaluation of dogs with marrow iron score <3 for external blood loss or nutritional deficiencies is likely clinically worthwhile, particularly if there is also decreased CHr.

Apoptosis in mosquito salivary glands: Sindbis virus-associated and tissue homeostasis.

Apoptosis is observed during a spectrum of conditions including exogenous virus infection and endogenous cellular turnover. Adult female Aedes albopictus mosquitoes challenged with increasing titres of Sindbis virus (SINV) via intrathoracic inoculation demonstrated that the injection dosage did not result in significantly different levels of virus growth or mosquito survival at day 10 post-infection. Tissues probed for apoptosis using an in situ TUNEL assay revealed SINV-associated apoptotic cells scattered throughout the proximal and distal regions of the salivary gland (SG) lateral lobes but which were not detected in the median lobe or the midgut and hindgut. Apoptosis was also identified in SG duct cells in both infected and uninfected mosquitoes, suggesting routine tissue homeostasis. SINV-associated apoptosis sequestered to the SG lateral lobes indicates a differential epithelial cell response to an arbovirus and provides insight into mosquito defence mechanisms against pathogens and SG infection barriers, hurdles to transmission of arboviruses of public health concern.