Classification of new BVDV isolates from Philippine water buffalo using the viral E2 region

Characterization of bovine viral diarrhea virus (BVDV) isolates has been focused of several studies this last decade. Until now lots of new strains are being unfolded maybe due to the viral fast mutation ability. As we focused our research on water buffalo immunology, we were able to identify a probable new BVDV isolates. RNA was extracted from water buffalo blood in the Philippines. The extracted RNA was reverse‐transcribed and synthesized cDNA. Oligonucleotide primers from the viral E2 region were used to amplify the target viral gene and later purified, cloned and sequenced. The E2 region with 420 bp nucleotides long was compared with existing published sequences in the GenBank. Based on the constructed phylogenetic tree, the isolated strain showed to be a BVDV type 1b along with Osloss and CP7 strains. Further classification of the new isolates was done within the BVDV type 1b1 group, which was compared with other strains in the sub‐group. The analysis revealed that Lamspringe/738, KE9 and 2543/87 were the closest with 92% homology. Additional study is being done to further qualify and quantify the extent of the existence of this new BVDV isolates in water buffalo in the Philippines. This is the first report of BVDV in the Philippines and first concerning BVDV in Philippine water buffalo. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

A case of solitary fibrous tumor of the kidney: an immunohistochemical and ultrastructural study with a review of the literature

A solitary fibrous tumor (SFT) is an unusual spindle cell neoplasm that usually arises in the pleura but rarely occurs in the kidney. Despite its rarity, histological diagnosis of SFT is crucial to avoid misdiagnosis with other malignant tumors in the kidney. We report a SFT of the left kidney that presented as a malignant tumor on radiographic findings in a 75-year-old Japanese woman. The tumor was well circumscribed and composed of a mixture of spindle cells and dense collagenous bands with no areas of necrosis or cystic changes noted macroscopically or microscopically. Electron microscopy showed fibroblast-like cells with well-developed rough endoplasmic reticulum, surrounded with collagen fibers. Immunohistochemistry revealed reactivity for vimentin, CD34, Bcl-2, and CD99, but no staining for cytokeratin, S-100, desmin, actin, D2-40, or epithelial membrane antigen (EMA). These findings were compatible with those of SFT. Although SFT of the kidney is extremely rare, this tumor must be included in the differential diagnosis when we encounter renal tumors consisting of mesenchymal elements. Immunohistochemical study is the key to diagnosis for SFT, and ultrastructural study is useful for its diagnosis.     

Epithelial Mesenchymal Transition in Human Ocular Chronic Graft-Versus-Host Disease

Chronic graft-versus-host disease (cGVHD) of the ocular surface and lacrimal gland is a vision-threatening condition that occurs after allogeneic bone marrow transplantation. In this study, we used immunohistochemistry and electron microscopy to investigate whether epithelial mesenchymal transition (EMT) contributed to the pathogenesis of ocular cGVHD. We detected down-regulation of E-cadherin and translocation of β-catenin from the intercellular junction to the cytoplasm and nucleus of cGVHD conjunctival basal epithelia and lacrimal gland myoepithelia. Notable findings included expression of Snail, an inducer of EMT, in the nucleus of ocular cGVHD epithelia. The fibrosis markers heat shock protein 47, α-smooth muscle actin, and fibroblast specific protein-1 were overexpressed in ocular cGVHD epithelia. In addition, p63, a marker of conjunctival basal epithelia, was observed in the nuclei of subconjunctival cells beneath disrupted basal lamina. Disrupted basal lamina and the presence of altered collagen bundles were observed in the cytoplasm and beneath cGVHD epithelia. In contrast, these observations were rarely observed in the normal conjunctiva and in Sjögren’s syndrome lacrimal gland epithelia. These findings together indicate that ocular cGVHD epithelia gain the mesenchymal phenotype and the capacity to migrate into the subepithelial stroma. Our findings suggest that EMT may be partially responsible for the conjunctival and lacrimal gland fibrosis found in patients with cGVHD.Allogeneic hematopoietic stem cell transplantation is a potentially curative treatment for hematological malignancies. However, chronic graft-versus-host disease (cGVHD) is a major complication after allogeneic hematopoietic stem cell transplantation (HSCT), which has hampered the success of HSCT.¹ Although numerous advances have been made for treating acute GVHD, the pathogenesis of cGVHD remains largely unknown and effective therapy has not been established. We previously studied the pathogenesis of cGVHD by focusing on the ocular surface and lacrimal gland and found that excessive fibrosis and a subset of fibroblasts contribute to the pathogenesis of ocular cGVHD.^2,3Recently, several studies have reported that epithelial-mesenchymal transition (EMT) contributes to various fibrotic diseases of the kidney,⁴ lung,⁵ and liver.⁶ For example, 40% of fibroblasts in kidney fibrosis arise from epithelial cells via local EMT triggered by inflammatory stress.⁷ EMT is involved in embryonic development, organ fibrosis, and also cancer metastasis.⁸ EMT is characterized by the loss of apical/basal cell polarity and loss of cell-to-cell adhesions, followed by the acquisition of a mesenchymal phenotype, ie, migration and invasion ability and expression of mesenchymal markers.EMT is triggered by various stimuli including irradiation,⁹ hypoxia,¹⁰ reactive oxygen species,¹¹ inflammatory cytokines such as transforming growth factor-β and fibroblast growth factor,⁸ disruption of basal lamina, and exposure of cytoplasm to extracellular matrix.¹² These triggers of EMT also participate in conjunction with the pathogenesis of cGVHD after HSCT. In a clinical setting, total body irradiation before HSCT and migrating inflammatory cells after HSCT generate substantial proinflammatory cytokines.¹ This “cytokine storm” then acts on T cells in the graft, prompting them to attack host antigens.¹³ In addition, reactive oxygen species-mediated organ injury was reported in bone marrow transplant recipients.¹⁴ The purpose of this study was to elucidate whether EMT is involved in mucosal and exocrine gland cGVHD.     

Accelerated induction of mycobacterial antigen-specific CD8+ T cells in the Mycobacterium tuberculosis-infected lung by subcutaneous vaccination with Mycobacterium bovis bacille Calmette–Guérin

Both CD4⁺ and CD8⁺ T cells are important in protection against Mycobacterium tuberculosis infection. To evaluate the effect of vaccination with Mycobacterium bovis bacille Calmette–Guérin (BCG) on the CD8⁺ T-cell response to pulmonary M. tuberculosis infection, we analyzed the kinetics of CD8⁺ T cells specific to the mycobacterial Mtb32a_309–318 epitope, which is shared by M. tuberculosis and M. bovis BCG, in the lung of mice infected with M. tuberculosis. The CD8⁺ T cells were detected by staining lymphocytes with pentameric major histocompatibility complex (MHC) class I H-2D^b–Mtb32a_209–318 peptide complex and were analysed by flow cytometry. Mtb32a-specific CD8⁺ T cells became detectable on day 14, and reached a plateau on day 21, in the lung of M. tuberculosis-infected unvaccinated mice. Subcutaneous vaccination with M. bovis BCG in the footpads induced Mtb32a-specific CD8⁺ T cells in the draining lymph nodes (LNs) on day 7 and their numbers further increased on day 14. When M. bovis BCG-vaccinated mice were exposed to pulmonaryinfection with M. tuberculosis 4 weeks after vaccination, the Mtb32a-specific CD8⁺ T cells in the infected lung became detectable on day 7 and reached a plateau on day 14, which was 1 week earlier than in the unvaccinated mice. The pulmonary CD8⁺ T cells from the BCG-vaccinated M. tuberculosis-infected mice produced interferon-γ in response to Mtb32a_209–318 peptide on day 7 of the infection, whereas those of unvaccinated mice did not. The results demonstrate that induction of mycobacterial antigen-specific protective CD8⁺ T cells in the M. tuberculosis-infected lung is accelerated by subcutaneous vaccination with M. bovis BCG.     

Suppression of cell proliferation and cytokine expression by HL-p36, a tick salivary gland-derived protein of Haemaphysalis longicornis

Previously, a putative immunosuppressant-coding gene was identified from a complementary DNA library derived from the salivary glands of partially-fed Haemaphysalis longicornis. Using real-time polymerase chain reaction, the gene was shown to be predominantly expressed during blood feeding with the site of expression being mainly in the salivary glands; this was confirmed by Western blotting analysis. To investigate the function of this novel protein, in this study, we examined the proliferative responses of bovine mononuclear cells and murine splenic cells as well as the expression of profiles of several cytokines in these cells in the presence of the recombinant protein (H. longicornis-derived 36 000 molecular weight protein: rHL-p36). The addition of rHL-p36 at the beginning of the 72 hr cultivation period clearly inhibited proliferation of several mitogen-stimulated cells in a dose-dependent manner, with concomitantly significant down-regulation of messenger RNA levels for interleukin-2. The inhibitory response could be abrogated by blockage of HL-p36 with antibody, suggesting the direct involvement of rHL-p36 in the cell proliferation. Furthermore, the proliferative response of splenocytes isolated from rHL-p36-inoculated mice was significantly lower than for those from control mice, suggesting that rHL-p36 could also directly suppress immune responses in vivo. Interestingly, microarray analysis of the splenocytes showed that the expression of several immunomodulating genes was down-regulated by rHL-p36 inoculation. In conclusion, these results suggest that HL-p36 is an immunosuppressor that might play an important role in the modulation of host immune responses.     

In the ear of the beholder: neural correlates of adaptation to voice gender

While high-level adaptation to faces has been extensively investigated, research on behavioural and neural correlates of auditory adaptation to paralinguistic social information in voices has been largely neglected. Here we replicate novel findings that adaptation to voice gender causes systematic contrastive aftereffects such that repeated exposure to female voice adaptors causes a subsequent test voice to be perceived as more male (and vice versa), even minutes after adaptation [S.R. Schweinberger et al. , (2008) , Current Biology, 18, 684–688). In addition, we recorded event-related potentials to test-voices morphed along a gender continuum. An attenuation in frontocentral N1–P2 amplitudes was seen when a test voice was preceded by gender-congruent voice adaptors. Additionally, similar amplitude attenuations were seen in a late parietal positive component (P3, 300–700 ms). These findings suggest that contrastive coding of voice gender takes place within the first few hundred milliseconds from voice onset, and is implemented by neurons in auditory areas that are specialised for detecting male and female voice quality.     

ETFDH mutations,CoQ10 levels,and respiratory chain activities in patients with riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency

Multiple acyl-CoA dehydrogenase deficiency (MADD) is a metabolic disorder due to dysfunction of electron transfer flavoprotein (ETF) or ETF-ubiquinone oxidoreductase (ETF-QO). Mutations in ETFDH, encoding ETF-QO have been associated with both riboflavin-responsive and non-responsive MADD as well as a myopathic form of CoQ₁₀ deficiency, although pathomechanisms responsible for these different phenotypes are not well-defined. We performed mutation analysis in four Taiwanese MADD patients. Three novel ETFDH mutations were identified in four patients and all harbored the p.A84T mutation. Muscle CoQ₁₀ levels and respiratory chain activities measured in two patients were normal. Three patients improved on riboflavin together with carnitine. Our results show that not all MADD patients have CoQ₁₀ deficiency. Based upon our data, riboflavin and carnitine may be the first-line treatment for MADD.     

A surgical case of descending colon cancer with abdominal wall abscess after drainage of abscess

We report a surgical case of descending colon cancer with abdominal wall abscess. A 72-year-old man was admitted to a hospital because of left lower abdominal mass with slight pain. An abdominal CT showed a left lower abdominal wall abscess adjacent to the descending colonic wall thickening. We diagnosed an abdominal wall abscess due to descending colon cancer or colon diverticulitis. The abscess was drained under local anesthesia releasing foul-smelling pus and air. After abscess drainage and general improvement in his condition, we conducted subtotal colectomy with lymph node dissection and excision of abdominal wall abscess cavity. Pathological findings indicated moderately differentiated adenocarcinoma of the descending colon (pT4, pN0, sH0, sP0, sM0, fStage II). The carcinoma had invaded the abdominal wall and transverse colon, but the cancer cells were not shown in the abdominal wall abscess cavity. In abdominal wall abscess treatment, colon cancer should be considered as a potential underlying cause. CT proved useful for assessing the status of the tumor and the abscess. We conducted a radical operation for descending colon cancer after the drainage for abdominal wall abscess.     

Aprepitant successful for controlling nausea and vomiting in a case having difficulty with control treatment in a standard antienemic therapy involving chemotherapy for gallbladder cancer postoperative recurrence

This case was a 62-year-old female. She underwent radical surgery for advanced gallbladder cancer 2 years ago after preoperative chemotherapy consisting of GEM/5-FU and CDDP (GFP). Two years after surgical treatment, multiple lung metastases and lymph node metastases appeared, and therefore, GFP chemotherapy was introduced. Rapid emesis occurred at two-cycle medication the first day, and was continued for several days. It was difficult to control the emesis by standard antienemic therapy. We therefore used aprepitant, a new medicine for antiemetic therapy. It had an excellent effect, and chemotherapy for this patient is still being continued.     

A case of toxicity caused by drug interaction between capecitabine and phenytoin in patient with colorectal cancer

We present a case of toxicity caused by a drug interaction between capecitabine and phenytoin (PHT). The drug combination elevated the plasma level of PHT in a patient on chemotherapy with capecitabine for colorectal cancer. Our patient was a 44-year-old woman diagnosed with epilepsy in her 20's, being treated with valproic acid (VPA) and PHT. Adjuvant chemotherapy with capecitabine began following surgery for colorectal cancer. Seven weeks later, she developed numbness, dizziness, dysarthria and difficulty walking, and was hospitalized for investigation. Her serum PHT level was elevated at 35. 1 μg/ mL. This case suggests that when capecitabine and PHT are coadministered, PHT levels should be monitored frequently, and that PHT dosage should be adjusted accordingly if it cannot be replaced by an alternative anticonvulsant.