Stromal-type Nephroblastoma with or without Anaplasia in Two Hedgehogs (Atelerix albiventris).

We describe the clinical and histological characteristics of stromal-type nephroblastomas that developed in two hedgehogs (Atelerix albiventris). In case 1, the tumour was composed of a proliferation of anaplastic stromal cells with ductal structures resembling the epithelium of nephroblastoma. In case 2, spindle-shaped cells that were somewhat larger than nephroblasts were frequently seen surrounding the cell cluster, and there was proliferation of stromal cells with collagen fibres at the periphery. Immunohistochemically, the tumour cells labelled weakly to strongly for the nephroblast marker Wilms' tumour-1 and were positive for Ki67 with rates of 5% and 10% for cases 1 and 2, respectively. Based on the above, the diagnosis was of stromal-type nephroblastoma with anaplasia in case 1 and without anaplasia in case 2. Our findings suggest that stromal-type nephroblastomas arise in adult hedgehogs and are clinically benign, and that histological anaplasia does not affect the prognosis.

CBCT location of the fusion between the buccal and lingual cortical in the mandibular ramus: importance to sagittal split osteotomy.

BACKGROUND: Mandibular Sagittal Split Osteotomy (MSSO) is a popular technique in orthognathic surgery used both to advance and to retreat the mandible. However, MSSO may incur in important complications, such as bad splits and sensorineural injuries. Knowing the location of the fusion between the buccal and lingual cortical (FBLC) in the mandibular ramus and the bone thickness in the region where osteotomies will be performed is determinant in MSSO planning to avoid complications. The aim of this study was to document and evaluate possible differences between sexes regarding the location of the FBLC in relation to the superior cortical of mandibular foramen (MF) and bone thickness in the region of interest for MSSO in a Brazilian population. MATERIAL AND METHODS: Eighty five cone-beam Computed Tomography (CBCT) scans were used to perform linear measurements to determine the location of the FBLC. Bone thickness from the mandibular canal (MC) to the cortical external surfaces and the diameter of the MC were measured at three different points: mandibular ramus (A), mandibular angle (B) and mesial of the second molar (C). RESULTS: The FBLC was located at a mean distance of 8.3 mm from the superior cortical of the MF in males and 8.1 mm in females. There was no difference between males and females regarding the mean bone thickness from the MC to the buccal external surface at all the points investigated (p>0.05). Bone thickness from the lingual external surface to the MC was bigger among females than males in regions B and C (p < 0.05). The diameter of the MC was bigger among males in regions B and C. CONCLUSION: Sexual dimorphism regarding mandibular bone thickness but not regarding the location of FBLC was present. This fundamental knowledge may assist to the panning of MSSO.

How to assess tonsilloliths and styloid chain ossifications on cone beam computed tomography images.

OBJECTIVE: The aim of this study was to establish an anatomical guideline in cone beam computed tomography (CBCT) images to discriminate soft-tissue calcifications, specifically, tonsilloliths, and styloid chain ossification (SCO) in the multiplanar reconstruction screen of the i-CAT Vision. MATERIALS AND METHODS: We analyzed 100 pairs of CBCT images and panoramic digital radiographies regarding the presence or absence of tonsilloliths and SCO. The intraobserver agreement varied from excellent to good. The statistical analyses included Mann-Whitney test, chi-square test, Spearman test, Student's t-test, and Wilcoxon test. The analyses were repeated without the guideline to verify its effectiveness. RESULTS: A total of 25 tonsilloliths were found in panoramic images while CBCT images revealed 60. Panoramic and CBCT images showed 42% and 63% of patients positive to SCO, respectively. We found a statistically significant difference when comparing the presence of tonsilloliths and SCO between panoramic and CBCT images (Wilcoxon test P < 0.05). The analyses without the guideline showed that the observer tended to diagnoses more false-positive SCO. CONCLUSION: Based on the results, we can suggest that CBCT images are more suitable to differentiate tonsilloliths and SCO than panoramic images. The guideline was more important to diagnosis SCO than tonsilloliths. SCO was misclassified in 34% without the guideline.

Effects of prolonged water washing of tissue samples fixed in formalin on histological staining.

The effects of prolonged water washing after fixation for 48 h in 10% (v/v) phosphate-buffered neutral formalin on the quality of representative histological staining methods were evaluated using samples of liver, kidney, spleen and thymus collected from three male Crl:CD(SD)(IGS) rats and one male beagle dog. Because door-to-door courier services in Japan prohibit handling formalin, our goal was to confirm that formalin fixed wet tissue samples could be stored in tap water rather than formalin during transportation of the samples without decreasing the quality of their staining or immunohistochemistry. Each tissue sample was allocated randomly to one of three groups: 12 min, 3 days and 7 days of washing in running tap water; samples then were routinely embedded in paraffin and sectioned. The sections were stained with hematoxylin and eosin, periodic acid-Schiff, azan, and the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. Immunohistochemical staining for Factor VIII, ED-1 and CD3 also was assessed. Prolonged water washing for up to 7 days did not affect the morphology or stainability by standard histological methods, or the intensity and frequency of positive reactions using the TUNEL method. Only immunohistochemical staining of Factor VIII was altered in both the rat and dog sections after 7 days of water washing. The intensity of positive reactions of Factor VIII immunohistochemistry after 7 days water washing was still strong enough to detect microscopically. Therefore, prolonged water washing for up to 7 days after formalin fixation does not have seriously detrimental effects on the quality and characteristics of paraffin sections stained by various methods, including immunohistochemistry.

Influence of hypobaric hypoxia on bispectral index and spectral entropy in volunteers.

BACKGROUND: Hypoxia has been shown to change electroencephalogram parameters including frequency and amplitude, and may thus change bispectral index (BIS) and spectral entropy values. If hypoxia per se changes BIS and spectral entropy values, BIS and spectral entropy values may not correctly reflect the depth of anaesthesia during hypoxia. The aim of this study was to examine the changes in BIS and spectral entropy values during hypobaric hypoxia in volunteers. METHODS: The study was conducted in a high-altitude chamber with 11 volunteers. After the subjects breathed 100% oxygen for 15 min at the ground level, the simulated altitude increased gradually to the 7620 m (25,000 ft) level while the subjects continued to breathe oxygen. Then, the subjects discontinued to breath oxygen and breathed room air at the 7620 m level for up to 5 min until they requested to stop hypoxic exposure. Oxygen saturation (SpO2), heart rate, 95% spectral edge frequency (SEF), BIS, response entropy (RE), and state entropy (SE) of spectral entropy were recorded throughout the study period. RESULTS: Of the 11 subjects, seven subjects who underwent hypoxic exposure for 4 min were analysed. SpO2 decreased to 69% at the 7620 m level without oxygen. However, SEF, BIS, RE, and SE before and during hypoxic exposure were almost identical. CONCLUSION: These data suggest that hypoxia of oxygen saturation around 70% does not have a strong effect on BIS and spectral entropy.

Clinical outcomes in older or younger patients with papillary thyroid carcinoma: impact of lymphadenopathy and patient age.

AIMS: To examine lymph node metastasis (LNM) from papillary thyroid carcinoma (PTC) according to clinicopathological features and outcomes associated with the nodal status. METHODS: We reviewed 231 patients with PTC (> or =1.0cm) who underwent initial thyroidectomy with modified neck dissection. LNM was examined in the central and lateral compartment and risk factors for disease-free survival (DFS) were evaluated. Nodal status and outcomes were further evaluated in four subgroups, 19 older patients (> or =45years old) with palpable lymphadenopathy (PLA) and 134 without PLA, and 11 younger patients (<45years old) with PLA and 67 without PLA, because multivariate analysis revealed that age (p<0.05, Hazard ratio (HR) 3.51) and PLA (p<0.0001, HR 14.9) were risk factors for DFS. RESULTS: Central and lateral LNM were found in 176 and 151 patients. Seventeen exhibited skip metastasis. Recurrence and disease death occurred in 23 and 5. In analysis of the four subgroups, recurrence was significantly frequent in older patients with PLA than in younger patients with PLA or older patients without PLA (8/19 vs. 3/11 or 12/134). Younger patients without PLA did not exhibit recurrence. CONCLUSIONS: Prognosis is worse in older patients with PLA. Such patients should be treated carefully with a considerable treatment strategy.

Partial protection against challenge with the highly pathogenic H5N1 influenza virus isolated in Japan in chickens infected with the H9N2 influenza virus.

The protective effect of the A/Ck/Yoko/aq55/01 (H9N2) avian influenza virus against the highly pathogenic H5N1 virus, i.e., A/Ck/Yama/7/04 (genotype V), was examined. Three 5-week-old chickens were inoculated intranasally with the H9N2 virus (10(8.6) EID(50)/head) and were kept with two contact chickens. All of the infected chickens were reinoculated with the same virus at 20 weeks of age, and 10 days later, they were challenged intranasally with the H5N1 virus (10(4.0) EID(50)/head). Five chickens simultaneously challenged with only the H5N1 virus (challenge control) died within 4 days postchallenge (d.p.c.). In contrast, four out of the five challenged, immune chickens died from 5 to 8 d.p.c. The median time to death in the immune chickens (6.3 days) was significantly longer than that in the challenge controls (3.4 days) (P < 0.01). No H5N1 virus shedding into the tracheae and feces of the challenged, immune chickens were detected for 3 d.p.c., but H5 genes were detectable in only one chicken by a loop-mediated isothermal amplification method. The H5N1 viruses were detected in the tracheae and/or feces of the dead immune chickens at death or 1 to 2 days before death. Only one out of the five challenged, immune chickens survived the H5N1 challenge without any signs for 14 d.p.c., but the virus and H5 gene were sporadically detected in the trachea only 7 and 14 d.p.c., respectively. This study shows that the H9N2 viruses may have the potential to induce cross-protection to the challenge with a recent lethal H5N1 virus (genotype V).

Impact of different husbandry conditions on contact and airborne transmission of H5N1 highly pathogenic avian influenza virus to chickens.

Typically highly pathogenic avian influenza (HPAI) viruses spread very rapidly among chickens within sheds. However, the spread was slower than expected for the initial 10 days of the index farm in Japan during 2004. This slow spread, as well as the lack of gross lesions, clinical signs, or high mortality, hindered the field veterinarian from reporting a suspected HPAI outbreak to the veterinary office. To understand the field conditions for the slow virus spread, we examined contact and airborne transmission of the H5N1 virus to chickens in a negative-pressure isolator using various numbers of infected chickens and separate compartments. We found that the contact transmission did occur inefficiently when one or two chickens were infected, whereas the transmission was efficient when four chickens were infected. Airborne transmission of the HPAI virus was also dependent on the number of infected chickens and was less efficient than contact transmission. These data together with field observations suggested that number of infected chickens, chicken house types, and amount of environmental contamination might affect the virus transmission efficiency to chickens.

Technique and assessment of sentinel lymph node biopsy usefulness in laparoscopy-assisted distal gastrectomy.

BACKGROUND: Recently, some studies have suggested that sentinel node biopsy also can be applied to gastric cancer. The authors apply sentinel lymph node biopsy in laparoscopy assisted distal gastrectomy to perform it as safe limited surgery. Limited surgery is a procedure in which the extent of lesion resection and lymph node dissection is reduced. The authors demonstrate that intraoperative diagnosis of lymph node metastasis is useful in this respect. METHODS: The study was conducted with 38 patients (29 men and 9 women) who had a preoperative diagnosis of T1 tumor invasion. The patients had a mean age of 66.2 years. Patent blue (1%) was injected submucosally into four or five different sites around the primary tumor at 1 ml per site. Blue-stained lymphatics and lymph nodes could be seen by turning over the greater omentum and the lesser omentum extraperitoneally. If blue nodes were found, biopsy was performed. RESULTS: The mean number of blue nodes dissected was 2.5 +/- 1.9. Intraoperative identification and biopsy of blue nodes could be performed for 35 (92.1%) of the 38 patients. Of the 35 patients in whom blue nodes were identified, 4 (9.7%) had metastases in blue nodes confirmed by intraoperative frozen-section diagnosis. Intraoperative frozen-section diagnosis was negative for blue node metastasis in 31 patients. Postoperative permanent section diagnosis also showed no evidence of lymph node metastasis in these 31 patients (100% accuracy, 0% false-negative rates). CONCLUSION: The reported method allows observation of blue-stained lymphatics up to 2 h after patent blue injection. Sentinel node biopsy was performed in laparoscopy assisted distal gastrectomy, making it technically equivalent to open gastrectomy. Sentinel node biopsy can serve as a method to determine the appropriate use of laparoscopy assisted distal gastrectomy for management of T1 gastric cancer.

Distribution of viral antigens and development of lesions in chicken embryos inoculated with nipah virus.

An isolate of Nipah virus was injected into fertile eggs via the allantoic cavity or yolk sac. Allantoic inoculation resulted in considerable pathological variation and only partial mortality. Dead embryos showed severe necrosis in the brain and congestion in the kidney and the subcutis of limbs. In contrast, yolk sac inoculation led to uniform infection and mortality, the dead embryos exhibiting the same lesions as those described above but without the subcutaneous congestion. Histological lesions in dead embryos inoculated by either route were similar and particularly severe in the central nervous system. Viral antigens were detected mainly in the vasculature and neurons. The results indicated that Nipah virus is highly pathogenic to chicken embryos, and that the route of inoculation is an important determinant of the course of disease. The findings also suggested that yolk sac inoculation can be used for viral titration, and that the chicken embryo represents a useful model for studying the vascular and neuronal tropisms of Nipah virus.

Pathology of fatal highly pathogenic H5N1 avian influenza virus infection in large-billed crows (Corvus macrorhynchos) during the 2004 outbreak in Japan.

Highly pathogenic H5N1 avian influenza viruses were isolated in 9 large-billed crows that died in Kyoto and Osaka prefectures in Japan from March to April in 2004. We studied 3 of the 9 crows using standard histologic methods, immunohistochemistry, and virus isolation. The most prominent lesions were gross patchy areas of reddish discoloration in the pancreas. The consistent histologic lesions included severe multifocal necrotizing pancreatitis, focal degeneration and necrosis of neuron and glial cells in the central nervous system, and focal degeneration of cardiac myocytes. All of these tissues contained immunohistochemically positive influenza viral antigens. The virus was isolated from the brain, lung, heart, liver, spleen, and kidney of the crows examined. Thus we concluded that highly pathogenic avian influenza virus was associated with clinical disease, severe pathologic changes, and death in the 3 crows.

Use of microcoil probehead for determination of the structure of oriented silk fibers by solid-state NMR.

A microcoil probehead for solid-state NMR was developed with a two-channel radio-frequency circuit, and 13C observation with a proton-decoupling probehead was performed to obtain information on the distribution of the orientation of silk fibroin molecules in the fiber. The coil (1 mm (diameter) x 5 mm (length)) of the probehead was placed at the angles 90 degrees and 30 degrees , relative to the static magnetic field. Only 70 mug of [1-13C]Gly silk fibroin fiber was used in a magnet of 9.4 T (400 MHz for proton channel).

Monoclonal antibody-based immunohistochemical diagnosis of Malaysian Nipah virus infection in pigs.

Formalin-fixed, paraffin wax-embedded tissues of three Malaysian farm pigs naturally infected with Nipah virus were used to investigate the value of anti-Nipah virus mouse monoclonal antibodies (Mabs) and rabbit polyclonal antibody for immunohistochemical diagnosis. Mabs 11F6 and 12A5 gave intense immunolabelling in lung tissue that had been fixed in 10% neutral buffered formalin for about 4 years, whereas the reactivity of Mabs 13A5 and 18C4 and polyclonal antibody was reduced significantly by long-term formalin fixation. Immunohistochemical examination of Malaysian farm pig samples with Mab 11F6 confirmed the affinity of Nipah virus for respiratory epithelium, renal glomerular and tubular epithelium, meningeal arachnoidal cells, and systemic vascular endothelium and smooth muscle. In addition, Nipah virus antigens were identified in laryngeal epithelial cells, Schwann cells of peripheral nerve fascicles in the spleen, and endothelial cells in the atrioventricular valve. The study demonstrated the value of Mabs 11F6 and 12A5 for the immunohistochemical diagnosis of Nipah virus infection in pigs.

[Clinical outcome of the operation using deep hypothermic cardiopulmonary bypass with intervals of circulatory arrest in thoracoabdominal aortic aneurysm].

Deep hypothermic cardiopulmonary bypass with intervals of circulatory arrest has been used for protection of the spinal cord during operations for thoracoabdominal aortic aneurysm (TAAA) in our hospital. We examined the effect of this adjunct this time. We studied 15 patients who were operated using deep hypothermic cardiopulmonary bypass with intervals of circulatory arrest among 19 patients with the TAAA who we performed the operations from 1995 through 2003. The patients ranged in age from 21 to 80 (an average of 65 +/- 14 SD) years. We used deep hypothermic cardiopulmonary bypass with intervals of circulatory arrest between 16 and 20 degrees C for the adjunct but did not use a monitor of evoked spinal cord potentials or cerebrospinal fluid drainage. Operation time was an average of 805 +/- 168 minutes. Cardiopulmonary bypass time was an average of 403 +/- 73 minutes. Deep hypothermic cardiopulmonary bypass time was an average of 215 +/- 67.5 SD minutes. Duration of spinal cord ischemia to the intercostal arteries were reconstructed was from 25 to 104 (50.5 +/- 24) minutes. We recognized nerve disorder in 6 cases in progress after operation, and respiratory organs management period and a hospitalization period became long, but the hospitalization death was 3 cases, and, as for the paraplegia was no case, 12 patients were discharged in good condition. The deep hypothermic cardiopulmonary bypass with intervals of circulatory arrest was regarded as a useful adjunct for prevention of the paraplegia.

Staple line coverage with absorbable mesh after thoracoscopic bullectomy for spontaneous pneumothorax.

BACKGROUND: Thoracoscopic simple bullectomy for primary spontaneous pneumothorax (PSP) has a relatively high postoperative recurrence rate and sometimes results in postoperative air leakage. One of the reasons for postoperative recurrence is the regrowth of bullae around the staple line. Therefore, reinforcement of the visceral pleura around the staple line is a reasonable way to prevent postoperative air leaks and recurrence. This study was done to determine the efficacy in preventing postoperative air leak and recurrent pneumothorax of widely covering the staple line with absorbable mesh after thoracoscopic bullectomy. METHODS: Wide coverage of the staple line with absorbable mesh was performed on 114 patients with PSP. These patients were retrospectively compared with 126 patients who underwent thoracoscopic simple bullectomy alone. RESULTS: The postoperative duration of chest drainage in the coverage group (mean, 1 day; range, 0-5) was significantly shorter than that in the simple bullectomy group (mean, 3 days; range 0-20). A prolonged air leak (>7 days) occurred in six patients in the simple bullectomy group, but there were no such leaks in the coverage group. Recurrent pneumothorax occurred in three patients (2.6%) in the coverage group and 12 patients (9.5%) in the simple bullectomy group. CONCLUSION: Wide coverage of the staple line with absorbable mesh is effective in preventing postoperative air leak and in decreasing the recurrence rates of PSP.

Development of breed identification markers derived from AFLP in beef cattle.

In the meat industry, correct breed information in food labeling is required to assure meat quality. Genetic markers provide corroborating evidence to identify breed. This paper describes the development of DNA markers to discriminate between Japanese Black and F1 (Japanese Black×Holstein) breeds. Amplified fragment length polymorphism method was employed to detect candidate markers absent in Japanese Black but present in Holstein. The 500 primer combinations yielded six selected markers that were converted into single nucleotide polymorphisms markers for high-throughput genotyping. The allele frequencies in both breeds were investigated for discrimination ability using PCR-RFLP. The probability of identifying F1 was 0.882 and probability of misjudgment was 0.0198. The markers could be useful for discriminating between Japanese Black and F1 and would contribute to the elimination of falsified breed labeling of meat.

Effect of sodium valproate on neuromagnetic responses to chromatic flicker: implication for photosensitivity.

The effect of oral administration of sodium valproate in normal subjects was evaluated using whole-scalp magnetoencephalography, with results compared to the effect of sodium valproate in photosensitive children. Neuromagnetic responses to 10 Hz equiluminant red-green and red-blue flicker were measured before and after 5 days of sodium valproate administration. For the red-green flicker, relative power spectra at the stimulus frequency (10 Hz) were attenuated with medication in most brain regions. However, for the red-blue flicker, the 10-Hz power in the occipital region was enhanced with medication, while it was reduced in other regions. These results qualitatively resembled those in photosensitive children. The present findings suggest that (1) combinational chromatic sensitivity can be a critical factor for cortical excitability, that (2) the effect of sodium valproate is qualitatively similar in normal and photosensitive subjects, and that (3) the effect of sodium valproate on cortical excitability is not simply to suppress the stimulus-synchronized occipital activity, but rather to inhibit the spread of cortical activity from the occipital region to other regions.

Pancreatic necrosis and ventricular erosion in adenovirus-associated hydropericardium syndrome of broilers.

Seven 19-day-old broiler chickens affected with hydropericardium syndrome (HPS) with pancreatic necrosis and gizzard erosions were investigated pathologically and virologically. Mortality increased after 13 days of age in a flock on a broiler farm. The mortality rate of the flock reached 10% by 19 days of age. Macroscopically, the chickens had hydropericardium (the characteristic gross change of HPS), pinpoint white foci in the pancreas, and ventricular erosions. Histologically, the chickens had multifocal hepatic necrosis with intranuclear inclusions in hepatocytes, a marked increase of macrophages in the spleen and lung, mild epicardial edema, multifocal necrosis of pancreatic acinar cells with intranuclear inclusions, focal necrosis of the ventricular koilin layer, and degeneration of the ventricular glandular epithelium with intranuclear inclusions. Immunohistochemically, intranuclear inclusions in the liver, pancreas, and ventriculi were stained positively against group I avian adenovirus (GIAAV) antigens. Ultrastructurally, 67-nm diameter viral particles were present in intranuclear inclusions. Virologically, serotype 4 of GIAAV was isolated from the liver, heart, and kidney of affected chickens. The pathologic changes of the present cases differ from previous cases of HPS; therefore, the present strain of GIAAV may have different pathogenicity for chickens than the previous virus strain of HPS.

Complete, long-lasting protection against lethal infectious bursal disease virus challenge by a single vaccination with an avian herpesvirus vector expressing VP2 antigens.

Marek's disease herpesvirus is a vaccine vector of great promise for chickens; however, complete protection against foreign infectious diseases has not been achieved. In this study, two herpesvirus of turkey recombinants (rHVTs) expressing large amounts of infectious bursal disease virus (IBDV) VP2 antigen under the control of a human cytomegalovirus (CMV) promoter or CMV/beta-actin chimera promoter (Pec promoter) (rHVT-cmvVP2 and rHVT-pecVP2) were constructed. rHVT-pecVP2, which expressed the VP2 antigen approximately four times more than did rHVT-cmvVP2 in vitro, induced complete protection against a lethal IBDV challenge in chickens, whereas rHVT-cmvVP2 induced 58% protection. All of the chickens vaccinated with rHVT-pecVP2 had a protective level of antibodies to the VP2 antigen at the time of challenge, whereas only 42 and 67% of chickens vaccinated with rHVT-cmvVP2 or the conventional live IBDV vaccine, respectively, had the antibodies. The antibody level of chickens vaccinated with rHVT-pecVP2 increased for 16 weeks, and the peak antibody level persisted throughout the experiment. The serum antibody titer at 30 weeks of age was about 20 or 65 times higher than that of chickens vaccinated with rHVT-cmvVP2 or the conventional live vaccine, respectively. rHVT-pecVP2, isolated consistently for 30 weeks from the vaccinated chickens, expressed the VP2 antigen after cultivation, and neither nucleotide mutations nor deletion in the VP2 gene was found. These results demonstrate that the amount of VP2 antigen expressed in the HVT vector was correlated with the vaccine efficacy against lethal IBDV challenge, and complete protective immunity that is likely to persist for the life of the chickens was induced.