Evaluation of 3-week-old layer chicks intratracheally challenged with Salmonella isolates from serogroup c1 (O:6,7) and Salmonella Enteritidis.

With the exception to Salmonella enterica serotype Typhimurium and S. Enteritidis (serogroups B [O:4] and D [O:9], respectively), there have been very few studies conducted on the respiratory tract as route of infection in chickens with salmonellas from serogroup C1 (O:6,7). Therefore, the purpose of this present study was to determine the potential organ invasion by Salmonella enterica serotype Potsdam (SP), S. Mbandaka (SM), and S. Infantis (SI) from serogroup C1 (O:6,7) and compare their characteristics with those of S. Enteritidis (SE) on intratracheally (IT) challenged 3-week-old layer chicks. A total of 360 one-day-old White Leghorn layer chicks were acquired from a commercial hatchery and randomly assigned into four treatment groups (SP, SM, SI, and SE, respectively), consisting of three independent trials. Chicks were grown up to 21 days (3 weeks) and IT-challenged thereafter with 106 CFU of respective salmonella organisms per group (n = 30). Chicks (n = 5) were humanely sacrificed every 24 h for 6 days post-IT infection and organs such as lung, heart, liver, spleen, kidney and caecal content were cultured for salmonella. All treatment groups exhibited colonization of lungs and caecal contents at 1 d (P = 0.475) and 4 d (P = 0.696) post-IT infection, respectively. There was no isolation of SP, SM, and SI in heart, liver, spleen, and kidney. In contrast, SE was obtained from heart, liver, and spleen of IT-infected chicks. The findings of this study contribute to a better understanding of the importance of the respiratory route in salmonella infection in poultry.

Characterization of Ornithobacterium rhinotracheale from commercial layer chickens in eastern Japan.

From a total of 72 commercial layer and pullet farms that were monitored in the eastern Japan area, 4 farms had mild to severe respiratory disease accompanied by decreased feed intake and drop in egg production. Microbiological analysis showed that 3 of the 4 farms, particularly from Fukushima, Tochigi, and Ibaraki prefectures, were positive for Ornithobacterum rhinotracheale (ORT). Out of 65 birds examined, ORT was isolated in 21 birds (32.31%). All isolates were Gram-negative pleomorphic rods with a colony size of 0.05 mm, translucent with grayish coloration, and with butyric smell after 48 h of incubation in 10% chicken blood agar at 37°C under microaerophilic conditions. All isolates reacted positively in the p-nitrophenyl-ß-d-galactopyranoside test within 3 h and were positive in cytochrome oxidase tests with an API 20NE identification system biocode of 0-0-2-0-0-0-4. An agar gel precipitation test showed that all isolates were serotype-A. All strains were positive in PCR by yielding a 784 bp amplicon of the 16S rRNA gene. All strains were resistant to amikacin, colistin, gentamicin, kanamycin, neomycin, polymyxin b, streptomycin, and sulfamethoxazole trimethoprim and susceptible to amoxicillin clavulanic acid, ampicillin, doxycycline, spectinomycin, and tetracycline. This study is the first characterization of ORT from commercial layer chickens in eastern Japan.

Identification of a Unique Amyloid Sequence in AA Amyloidosis of a Pig Associated With Streptococcus Suis Infection.

Here we report a pig with amyloid A (AA) amyloidosis associated with Streptococcus suis infection and identification of a unique amyloid sequence in the amyloid deposits in the tissue. Tissues from the 180-day-old underdeveloped pig contained foci of necrosis and suppurative inflammation associated with S. suis infection. Congo red stain, immunohistochemistry, and electron microscopy revealed intense AA deposition in the spleen and renal glomeruli. Mass spectrometric analysis of amyloid material extracted from the spleen showed serum AA 2 (SAA2) peptide as well as a unique peptide sequence previously reported in a pig with AA amyloidosis. The common detection of the unique amyloid sequence in the current and past cases of AA amyloidosis in pigs suggests that this amyloid sequence might play a key role in the development of porcine AA amyloidosis. An in vitro fibrillation assay demonstrated that the unique AA peptide formed typically rigid, long amyloid fibrils (10 nm wide) and the N-terminus peptide of SAA2 formed zigzagged, short fibers (7 nm wide). Moreover, the SAA2 peptide formed long, rigid amyloid fibrils in the presence of sonicated amyloid fibrils formed by the unique AA peptide. These findings indicate that the N-terminus of SAA2 as well as the AA peptide mediate the development of AA amyloidosis in pigs via cross-seeding polymerization.

Expression of Periostin in Normal, Atopic, and Nonatopic Chronically Inflamed Canine Skin.

In humans, periostin plays a critical role in the enhancement and chronicity of allergic skin inflammation; however, whether it is involved in the pathogenesis of canine dermatitis remains unknown. The aim of this study was to examine the expression patterns of periostin in healthy, atopic, and nonatopic chronically inflamed canine skin. Biopsy specimens from 47 dogs with skin disease and normal skin tissue from 5 adult beagles were examined by light microscopy, immunohistochemistry, and in situ hybridization. In normal skin, periostin was localized just beneath the epidermis and around the hair follicles. In chronically inflamed skin, periostin expression was most intense in the dermis with inflammatory cell infiltrates. In contrast, low levels of periostin were detected in acutely inflamed and noninflamed skin. Conversely, all canine atopic dermatitis tissues characteristically showed the most intense expression of periostin in the superficial dermis, particularly at the epidermal-dermal junction. In situ hybridization showed that periostin mRNA was broadly expressed in the basal epidermal keratinocytes, outer root sheath cells, and dermal fibroblasts in normal dog skin. High expression of periostin mRNA was observed in fibroblasts in dog skin with chronically inflamed dermatitis. Moreover, in some chronically inflamed skin specimens, periostin mRNA expression was increased in basal keratinocytes. The severity score of chronic pathologic changes and CD3+ cell number in the dermis were correlated with distribution pattern of periostin in the atopic skin. These data suggest that periostin could play a role in the pathophysiology of chronic dermatitis, including atopic dermatitis, in dogs.

Proliferative lesions of intra-epidermal cytokeratin CAM5.2-positive cells in canine nipples.

Non-keratinocyte cells with clear or vacuolated cytoplasm are frequently observed in the epidermis of canine nipples. Most of these cells express cytokeratin (CK) CAM5.2, a marker of luminal epithelial cells. The morphological and immunohistochemical characteristics of these clear cells were investigated. Nipple tissue from 36 dogs of both sexes was collected and labelled immunohistochemically for CAM5.2, CK7, CK14, CK18, CK20, α-smooth muscle actin, p63, melan-A, E-cadherin, epidermal growth factor receptor and oestrogen receptor (OR). The intra-epidermal CAM5.2(+) clear cells were present singly or as small clusters, mostly within the basal layer, in 22 dogs (61%). These cells also expressed CK7, CK18, E-cadherin and OR. Electron microscopy revealed that some of these cells had surface microvilli. Multifocal proliferative lesions consisting of these cells were observed in the nipples of four dogs. In these lesions, proliferating cells formed bilayered tubules with CAM5.2(+) inner and CK14/p63(+) outer cells. This is the first report describing intra-epidermal CAM5.2(+) clear cells, distinct from melanocytes and Merkel cells in dog nipples. These cells might arise from the luminal epithelium of the papillary duct.

Development of podocyte injuries in Osborne-Mendel rats is accompanied by reduced expression of podocyte proteins.

Osborne-Mendel (OM) rats spontaneously develop glomerulopathy with progressive podocyte injury. Changes in protein expression levels in the foot processes of podocytes have been suggested to play an important role in the development of renal disease. The aim of this study was to investigate the temporal relationship between the expression of five podocyte proteins (nephrin, podocin, synaptopodin, α-actinin-4 and α3-integrin) and the development of podocyte injuries, proteinuria and glomerulosclerosis in OM rats. Male OM rats 5-20 weeks of age and age-matched Fischer 344 rats were used. Semiquantitative analysis of expression of the five podocyte proteins was performed by immunofluorescence labelling. Nephrin mRNA expression was determined by quantitative real-time reverse transcriptase polymerase chain reaction and nephrin protein expression was determined by mass spectrometry. Progressive reduction in expression of the podocyte proteins correlated with the progression of podocyte injuries, the development of proteinuria and the subsequent development of glomerulosclerosis. Nephrin mRNA expression and nephrin concentration also showed temporal decreases in OM rats. Altered expression of podocyte proteins preceded the development of proteinuria and glomerulosclerosis, suggesting that this event contributes to podocyte dysfunction and progression to glomerulosclerosis.

Expression of nephrin, podocin, α-actinin-4 and α3-integrin in canine renal glomeruli.

The biological features of podocytes that contribute to the pathogenesis of proteinuria have not been investigated in dogs. The aim of this study was to investigate the expression and localization of nephrin, podocin, α-actinin-4 and α3-integrin in canine renal glomeruli. Renal cortical tissue was collected from the kidneys of five normal adult beagles. Western blotting and immunofluorescence microscopy revealed specific expression and localization of the four proteins in canine glomeruli. Expression of genes encoding the four molecules in isolated glomeruli was detected by reverse transcriptase polymerase chain reaction. The results of this study will permit future exploration of podocyte injury and its involvement in protein leakage from the capillary wall in canine glomerular diseases.

Cardiac angioleiomyoma in 44 cattle in Japan (1982-2009).

A retrospective study was performed on primary cardiac tumors found in 44 cattle during meat inspection in Japan, using histology, immunohistochemistry, and electron microscopy. The age of affected cattle ranged from 10 to 129 months; 82% were less than 3 years old. In 38 hearts, the tumor was solitary; the remaining 6 hearts had multiple masses. All tumors were subendocardial and involved the papillary muscles and/or cardiac valves. Two histologic patterns were recognized; both included giant cells. Pattern 1 was characterized by interlacing fascicles of spindle-shaped cells; pattern 2 resembled cavernous hemangioma. The 2 patterns coexisted and were linked by transitional zones. Given the immunohistochemical reactivity and ultrastructure, the spindle-shaped cells were identified as smooth muscle cells and the giant cells as endothelial cells. Single cell and multicellular angiogenesis by giant cells was observed. The tumors were classified as cardiac angioleiomyoma. Subendocardial multipotential mesenchymal cells, persistent tissue of the endocardial cushion, or valvular interstitial cells were suspected as the origin.

[Coronary artery fistula into the right atrium complicated by atrial futter; report of a case].

A 62-years-old woman with uncommon atrial flutter underwent the catheter ablation therapy. In the catheter ablation therapy and subsequent coronary angiography, a coronary artery fistula into the right atrium was found. A giant aneurysm formed in the junctional region between the coronary artery fistula and right atrium. The giant aneurysm might have been a course of uncommon atrial flutter. We ligated the coronary artery fistula to prevent the giant aneurysm rupture. Any events have not been observed after catheter ablation and ligation.

Collagenofibrotic glomerulonephropathy with fibronectin deposition in a dog.

We report herein a case of collagenofibrotic glomerulonephropathy in a 3-year-old Shiba Inu with severe proteinuria. Histologically, renal glomeruli were enlarged with massive deposition of a homogeneous eosinophilic substance within the mesangium and capillary walls. The deposits reacted weakly with periodic acid-Schiff, stained deep blue with Masson's trichrome, and were positive by immunofluorescence for type III collagen and fibronectin. Ultrastructurally, the deposits consisted of fibrils and amorphous material in the mesangial matrix and beneath the glomerular capillary endothelium. The fibrils had transverse bands analogous to those of collagen fibrils. Electron microscopy also revealed focal detachment of podocytes and foot process effacement in glomerular tufts, which suggested that podocyte injury had contributed to the development of proteinuria in this dog. The current case resembles collagenofibrotic glomerulonephropathy (CFGN) in humans in histopathologic, immunofluorescence, and electron microscopic findings. This is the first report of CFGN in a nonhuman species with glomerular deposition of fibronectin and type III collagen.

Diagnostic exercise: Tyzzer's disease, distemper, and coccidiosis in a pup.

A 2-month-old mongrel dog had multifocal necrotizing hepatitis, interstitial pneumonia, and hemorrhagic enteritis. Immunohistochemistry detected antigens of Clostridium piliforme in the intestine and liver, and antigens of canine distemper virus within the lung, urinary bladder, brain, spleen, and liver. Furthermore, uncharacterized intralesional coccidian protozoa were observed within the intestine.

The role of roof rats ( Rattus rattus) in the spread of Salmonella Enteritidis and S. Infantis contamination in layer farms in eastern Japan.

The prevalence of Salmonella in four layer farms in eastern Japan was investigated between 2004 and 2006 to determine the role of roof rats (Rattus rattus) in the epizootology of Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis). Persistent S. Enteritidis and S. Infantis contamination of the environment and pooled egg samples were detected in three out of four layer farms. A total of 113 (13.3%) and 158 (18.6%) out of 851 rats examined were positive for S. Enteritidis and S. Infantis, respectively. By pulsed-field gel electrophoresis, only one indistinguishable pulsed-field pattern was yielded by S. Enteritidis strains from rats, eggs and environmental samples from each of the two contaminated layer farms. Although, a variety of pulsed-field patterns were generated by S. Enteritidis isolates from rats, eggs, and the environment of the other contaminated farms, there are, however, some S. Enteritidis strains that are closely related clones. These results suggest that roof rats are carriers of S. Enteritidis and S. Infantis and that persistent S. Enteritidis and S. Infantis infections in a rat population may play an important role in the spread and maintenance of these pathogens inside the layer premises.

[Radical operation of tetralogy of Fallot with left single coronary and long-term follow up; report of a case].

We report a long-term result of a radical repair of tetralogy of Fallot combined with left single coronary in 57-year-old woman. She was in severe respiratory failure and severe cyanosis. The repair was monocusp patch reconstruction of right ventricle (RV) out flow tract after cutting off the right coronary artery (RCA) and coronary artery bypass graft (CABG) to RCA with saphenous vein graft. The post operative course for these 11 years, she had been in controllable congestive heart failure from postoperative pulmonary regurgitation and myocardial ischemy owing to obstract of RCA graft.

Effects of reduction of the number of primordial follicles on follicular development to achieve puberty in female rats.

Effects of reduction of the number of primordial follicles on follicular development and concentrations of circulating hormones were examined in immature female rat offspring of dams given busulfan intraperitoneally on day 14 of gestation. The offspring of dams treated with 5 mg busulfan kg(-1) showed vaginal opening at an age comparable with the offspring of dams treated with 2.5 mg busulfan kg(-1) or with corn oil as a control, although they exhibited an irregular oestrous cycle until week 14 after birth. The serum concentrations of immunoreactive inhibin and FSH on day 26 after birth of the offspring treated with 5 mg busulfan kg(-1) were similar to those of age-matched controls. On day 15 after birth, however, the concentration of their immunoreactive inhibin was markedly lower than that of controls, whereas the concentration of their FSH was increased inversely. Comparison of the numbers of ovarian follicles in the controls and groups treated with 2.5 mg busulfan kg(-1) and 5 mg busulfan kg(-1) revealed that prenatal treatment with busulfan reduced the number of follicles in the primordial or primary phase and in the preantral phase on day 7 after birth. Although the increase of the ratio of the number of preantral follicles during days 7-13 after birth tended to vary with the prenatal dose of busulfan, the number of preantral follicles in the group treated with 5 mg busulfan kg(-1) was still smaller than in the controls. The concentration of serum immunoreactive inhibin of the offspring treated with busulfan was reduced on day 7 after birth without alteration of the concentration of gonadotrophin. On day 13 after birth, the concentration of serum immunoreactive inhibin was reduced only in the offspring treated with 5 mg busulfan kg(-1), and the concentration of serum FSH of the offspring was increased inversely as found on day 15 after birth. These results indicate that a reduction in the number of primordial follicles decreases the number of follicles that enter the growing phase, a major source of circulating inhibin in the neonatal and infantile ovary, and that consequently increased circulating FSH may accelerate follicular development to achieve puberty.

Characterization of novel kidney-specific delivery system using an alkylglucoside vector.

The alkylglucoside vector has been demonstrated to be a kidney-specific drug delivery system via cell surface-specific binding sites. In the present study, we examined the targeting efficiency of this vector derivatized with several types of ligand to determine the efficacy and limitations of this system. The tissue uptake clearance in the kidney (CL(uptake, kidney)) of alkylglucoside-acylated poly-L-lysine conjugates (Glc-S-C8-APL) with a mol. wt. of 4,500, 17,000, or 41,000 was greater than that accounted for by glomerular filtration and was reduced by coadministration of n-octyl-thioglucoside, which has an affinity for alkylglycoside binding sites. The mol. wt. distribution, assessed by gel filtration high-performance liquid chromatography, of the radioactivity associated with the kidney after intravenous administration of Glc-S-C8-APL41000 was shifted to a lower mol. wt. range compared with the authentic compound. The CL(uptake, kidney) and specific binding of Glc-S-C8-APL, fractionated based on mol. wt., to kidney membrane fractions was reduced as the mol. wt. of the fractionated Glc-S-C8-APL increased. These results suggest that the target efficiency of this vector depends on the size of the ligand that it delivers. Both the CL(uptake, kidney) and specific binding to kidney membranes of an alkylglucoside-tyrosine conjugate (Glc-S-C8-Tyr) with an acidic charge was much lower than that of Glc-S-C8-Tyr with cationic and neutral charges, suggesting that the anionic moiety could reduce the renal targeting efficiency. Thus, the targeting efficacy of the alkylglucoside vector seems to depend on, at least, the size and charge of the ligand that it delivers.

Experimental chemotherapy against canine mammary cancer xenograft in SCID mice and its prediction of clinical effect.

The effectiveness of 6 antitumor agents has been evaluated for canine mammary gland tumor (CMG-6) serially transplanted into severe combined immunodeficiency (SCID) mice. CMG-6 diagnosed as a solid carcinoma was subcutaneously transplanted into SCID mice and six antitumor agents were intravenously given to the mice as a single injection. The effectiveness was evaluated by Treatment group/Control group percent (T/C %) and statistical significance determined by Mann-Whitney's U-test in tumor volume. The minimum effective doses (MEDs; mg/kg) of mice were as follows; cyclophosphamide (CPM) 65, doxorubicin (DXR) 6, cisplatin (CDDP) 5, vincristine (VCR) 1.6, vinblastine (VLB) more than 5.5, 5-fluorouracil (5-FU) 105. Clinical effects of the drugs were predicted based on area under the curve (AUC) of dogs given a clinical dose (AUCdog)/AUC of mice given a MED (AUCmouse) ratios from published references. The AUC ratios were as follows; CPM 2.24, DXR 0.19, CDDP 1.20, VCR 0.04, VLB <1.24 and 5-FU 1.15. Drugs indicating more than 1.0 in AUCdog/AUCmouse ratio were CPM, CDDP and 5-FU, and would be suggested as effective in the original patient with CMG-6. The combination chemotherapy using clinically equivalent doses in CDDP and CPM, which were the two highest values in AUCdog/AUCmouse ratio by single agent therapy, was performed and shown to have additional effects as compared to the responsiveness of each agent against CMG-6.

Glucose alters the susceptibility of mesangial cells to contrast media.

RATIONALE AND OBJECTIVES: Diabetic patients frequently suffer contrast media-induced nephropathy. Hyperglycemia in diabetes mellitus causes gradual deterioration of glomerular mesangial cells (MCs) in the kidney. In this study, the authors investigated the response of rat MCs cultured in high-glucose medium to diatrizoate and iohexol, high- and low-osmolar contrast media, respectively. METHODS: Isolated rat MCs were precultured under basal-glucose (5.5 mmol/L) and high-glucose (30 and 55 mmol/L) conditions for 24 hours to mimic hyperglycemia in diabetes mellitus and then were exposed to diatrizoate (40 and 80 mg I/mL) and iohexol (80, 120, 160 mg I/mL) for 2 hours. The cytotoxic effects of diatrizoate and iohexol were monitored by neutral red uptake in MCs. The protective effects of an antioxidant, d-alpha-tocopherol (Toc), on cytotoxicity of the contrast media were determined when MCs were precultured with Toc under high-glucose conditions or were exposed to the contrast media together with Toc. Peroxide levels in the cells exposed to the contrast media were analyzed by flow cytometry using dichlorofluorescin diacetate. RESULTS: Exposure to both contrast media (diatrizoate and iohexol) induced a concentration-dependent decrease in viability of the cells precultured under basal-glucose conditions (5.5 mmol/L). Preculture under high-glucose conditions (30 and 55 mmol/L) augmented the cytotoxic effects of both contrast media. An increase in the intracellular peroxide level was detected after exposure to both contrast media. Preculture with Toc prevented augmentation of the cytotoxic effects of diatrizoate by the higher glucose concentration (55 mmol/L). The exposure to diatrizoate together with Toc also attenuated its cytotoxic effects. Toc showed no such protective effects against iohexol exposure. CONCLUSIONS: These findings suggest that high-glucose conditions enhance the susceptibility of MCs to the cytotoxic effects of both contrast media; the enhanced susceptibility was in part attributable to oxidative stress caused by high-glucose conditions; diatrizoate exerted the cytotoxic effects by means of oxidative stress; and iohexol appeared to exert its cytotoxicity in a manner different from diatrizoate.

Atypical membranoproliferative glomerulonephritis in a cat.

Membranoproliferative glomerulonephritis was observed in a 2-year-old male Japanese domestic cat with clinical renal failure. In the glomeruli, moderate mesangial hypercellularity with an increased mesangial matrix and thickening of the capillary walls were prominent. In addition, frequent duplication of the capillary walls, splitting, and spike formation were observed in the glomerular basement membrane. Granular cat IgG and complement component deposition were detected globally along the glomerular capillary walls and in the mesangium. Transmission electron microscopy revealed dense deposits in the subendothelial and subepithelial regions and the mesangium. Mesangial interposition was also observed. These glomerular lesions are also found in humans with membranoproliferative glomerulonephritis type III, which has not been reported in animals.

Auditory detection of simulated crackles in breath sounds.

BACKGROUND: Computerized analysis of breath sounds has relied on human auditory perception as the reference standard for identifying crackles. In this study, we tested the human audibility of crackles by superimposing artificial clicks on recorded breath sounds and having physicians listen to the recordings to see if they could identify the crackles. OBJECTIVES: To establish the audibility of simulated crackles introduced in breath sounds of different intensity, to study the effects of crackle characteristics on their audibility, and to investigate crackle detection within and between observers. METHODS: Fine, medium, and coarse crackles with large and small amplitude were synthesized by computer software. Waveform parameters were based on published characteristics of lung sound crackles. The amplitude for small crackles was defined as just above the threshold of audibility for simulated crackles inserted in sound recorded during breath hold. Simulated crackles were then superimposed on breath sounds recorded at 0 L/s (breath hold), 1 L/s, and 2 L/s airflow. Five physicians listened during playback on two separate occasions to determine if crackles could be heard and to calculate the interobserver and intraobserver variations. RESULTS: Failed detection of crackles was significantly more common in the following conditions: (1) background breath sounds had higher intensity (2 L/s airflow) compared to lower intensity (1 L/s), (2) crackle type was coarse or medium compared to fine, and (3) crackle amplitude was small compared to large. Both intraobserver and interobserver agreements were high (kappa > 0.6). RELEVANCE: The validation of automated techniques for crackle detection in lung sound analysis should not rely on auscultation as the only reference. Detection of crackles is facilitated when patients take slow, deep breaths that generate little breath sounds.