Clinicopathological study of experimentally induced canine monocytic ehrlichiosis

The aims of this study are to investigate the hematology, blood chemistry, pathological study, including macroscopic and microscopic lesions, of experimentally induced canine monocytic erhlichiosis in Thailand and to demonstrate the distribution of Ehrlichia canis in target organs by nested polymerase chain reaction (PCR). Five experimental healthy dogs were inoculated with 5 ml of whole blood (estimated number of E. canis morulae 15 × 10^–5% per monocytes) from the splenectomized dog via the saphenous vein. Two healthy dogs served as a negative control. Hematology revealed nonregenerative normocytic normochromic anemia, thrombocytopenia and mild leukopenia. Blood chemistry revealed an increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (AP), hypoproteinemia, hypoalbuminemia, and hyperglobulinemia by day 66 post-inoculation. Pathology revealed anemia, ascites, jaundice, interstitial pneumonia, splenomegaly, generalized lymphadenopathy, and severe fatty liver. The detection of E. canis was performed using tissue embedded in paraffin wax by nested PCR showing positive in all target organs. This study concluded that acute induced experimental canine monocytic ehrlichiosis can cause significant clinical and pathological lesions.     

Inter-sarcomere coordination in muscle revealed through individual sarcomere response to quick stretch

The force generation and motion of muscle are produced by the collective work of thousands of sarcomeres, the basic structural units of striated muscle. Based on their series connection to form a myofibril, it is expected that sarcomeres are mechanically and/or structurally coupled to each other. However, the behavior of individual sarcomeres and the coupling dynamics between sarcomeres remain elusive, because muscle mechanics has so far been investigated mainly by analyzing the averaged behavior of thousands of sarcomeres in muscle fibers. In this study, we directly measured the length-responses of individual sarcomeres to quick stretch at partial activation, using micromanipulation of skeletal myofibrils under a phase-contrast microscope. The experiments were performed at ADP-activation (1 mM MgATP and 2 mM MgADP in the absence of Ca²⁺) and also at Ca²⁺-activation (1 mM MgATP at pCa 6.3) conditions. We show that under these activation conditions, sarcomeres exhibit 2 distinct types of responses, either “resisting” or “yielding,” which are clearly distinguished by the lengthening distance of single sarcomeres in response to stretch. These 2 types of sarcomeres tended to coexist within the myofibril, and the sarcomere “yielding” occurred in clusters composed of several adjacent sarcomeres. The labeling of Z-line with anti-α-actinin antibody significantly suppressed the clustered sarcomere “yielding.” These results strongly suggest that the contractile system of muscle possesses the mechanism of structure-based inter-sarcomere coordination.     

Expression of E-cadherin, and CD44s and CD44v6 and its association with prognosis in head and neck cancer

OBJECTIVES: In the current study, the expression of E-cadherin, CD44s, and CD44v6 has been noted as markers for tumor metastasis and prognosis in several tumors, so we examined whether or not E-cadherin, CD44s, and CD44v6 are useful markers for evaluating the prognosis of mesopharyngeal cancer patients. METHODS: The expression of E-cadherin, CD44s, and CD44v6, was evaluated immunohistochemically using monoclonal antibodies against epitopes of standard and variant proteins, in paraffin-embedded mesopharyngeal cancer tissues from 57 patients who had received curative therapy. RESULTS: Tumor tissues from 47 (82.5%) patients showed positive immunoreactivity with monoclonal antibody against E-cadherin, 43 (75.4%) patients showed positive expression with CD44, and 45 (78.9%) patients showed positive expression with CD44v6. The expression of CD44v6 was slightly correlated with tumor volume, and lymph node metastasis, and stage classification (P > 0.05). However, there was no significant correlation between the expression of E-cadherin, CD44s and CD44v6 and clinicopathological characteristics. Concerning the prognosis, the survival period of patients with CD44s positive tumors was shorter than that of patients with CD44s negative tumors (18.2% versus 52.1%, 5-year survival, P > 0.05). The survival period of patients with CD44v6 positive tumors was also shorter than that of patients with CD44v6 negative tumors (12.8% versus 55.6%, 5-year survival, P > 0.05). CONCLUSION: These results suggest that CD44v6 may be related to tumor invasion and metastasis, and both CD44s and CD44v6 may be useful markers for poor prognosis in head and neck cancer.     

Nedaplatin for recurrent cancer of the head and neck

This study was undertaken to evaluate the clinical efficacy and toxicity of Nedaplatin (254-S) alone or combined for UFT for recurrent head and neck cancers in an outpatient setting. Thirty-two patients, previously treated, (30 men and 2 women, mean age 59 years, twenty one with loco-regional recurrence and 11 with distant metastasis, 29 with squamous cell carcinoma, 2 with adenocarcinoma and one with adenoid cystic carcinoma) were treated with Nedaplatin (254-S) alone or combined with UFT. The primary site was identified in the oropharynx in 8 patients, oral cavity in 7, larynx in 5, nasopharynx in 4, hypopharynx in 3, sinuses in one, parotid in one, and unknown primary in one patient. 254-S was administered at 80 mg/m2 by intravenous drip infusion. The 254-S administration was repeated at 4 week intervals, and in some patients was combined with daily oral administration of 400 mg of UFT-E (tegafur-uracil enterogranules). Twelve patients received 254-S alone and in 20 patients it was combined with UFT-E. The 254S administration ranged from one to 18 courses (mean of 5.7 courses). Grade 3-4 toxicities included leukopenia in 15.6%, anemia in 6.3% and thrombocytopenia in 9.4% of the patients. There was one death due to grade 4 leukopenic pneumonia. Four (12.5%) had a clinical complete and partial response. One-year and two-year overall survival rates were 35.6% and 30.5% for loco-regional recurrence, respectively. Ten of the eleven patients with distant metastasis died within six months and all patients were dead within 18 months, so a significant difference was observed in the overall survival rate between loco-regional recurrence and distant metastasis. Treatment with 254-S alone or combined UFT-E could be conducted in an outpatient setting and was able to improve the overall survival rate for recurrent head and neck cancer.     

Increased Protein-Conjugated Acrolein and Amyloid-β40/42 Ratio in Plasma of Patients with Mild Cognitive Impairment and Alzheimer's Disease

The objective of this study was to determine whether plasma levels of acrolein, a compound that causes cell damage, and amyloid-β (Aβ) are useful biochemical markers for Alzheimer's disease (AD). The study included 221 elderly subjects divided into 101 non-demented [33 healthy control and 68 non-demented subjects with white matter hyperintensity (nd-WMH)], 50 mild cognitive impairment (MCI), and 70 AD. Increases in both protein-conjugated acrolein (PC-Acro) and Aβ40/42 ratio were observed in MCI and AD patients compared with values in control subjects. When the combined measurements of PC-Acro and Aβ40/42 ratio were evaluated using the median value of the relative risk value for dementia, they were in the order AD (0.98) ≥ MCI (0.97) > nd-WMH (0.83) > control (0.35). The results indicate that measurements of PC-Acro and Aβ40/42 ratio not only detect MCI and AD patients but also nd-WMH subjects. Furthermore, both PC-Acro and Aβ40/42 ratio in plasma for 120 MCI and AD patients were significantly higher than those for 101 control and nd-WMH subjects, indicating that both values become useful biochemical markers for MCI and AD subjects.     

Application of quantitative gene expression analysis for pertussis vaccine safety control

Although vaccines are routinely used to prevent infectious diseases, little is known about the comprehensive influences caused by vaccines. In this study, we showed, using comprehensive gene expression analysis, that pertussis vaccine affected many genes in multiple organs of vaccine-treated animals. In particular, lung was revealed to be the most suitable target to evaluate pertussis vaccine toxicity. The 13 genes identified from the analysis of vaccine-treated lung at day 1 showed a clear dendrogram corresponding to pertussis vaccine toxicity. Furthermore, quantitative analysis of these genes revealed a positive correlation between their respective expression levels and the degree of toxic effects observed in samples that had been treated with various doses of reference pertussis vaccines. The quantification of this 13 gene-set is an indicator of the vaccine toxicity-related reaction.     

Lack of Direct Involvement of 8-Hydroxy-2'-deoxyguanosine in Hypoxanthine-guanine Phosphoribosyltransferase Mutagenesis in V79 Cells Treated with N,N'-Bis(2-hydroxyperoxy-2-methoxyethyl)-l,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or Riboflavin

The object of this study is to investigate the relationship between a typical product of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8OHdG), and mutagenesis in V79 cells through a molecular analysis of hypoxanthine-guanine phosphoribosyltransferase ( hprt ) gene mutants. We performed a direct sequencing analysis of the cDNA of mutants obtained after treatment with N,N'-bis(2-hydroxyperoxy-2-methoxyethyl)-l,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin, each of which induces the formation of 8OHdG in cellular DNA upon UVA irradiation. The frequency of mutation after both treatments was no more than 2 to 5 times the control value. A considerable number of the mutants could not be amplified by RT-PCR, and this was also the case for the control mutants. Among the mutants analyzed, deletions and a TA→Ã transversion occurred predominantly. The reasons for the weak association of induction of 8OHdG with frequency of mutation and the possible mechanism of oxidative-stress-derived mutagenesis are discussed.     

Clinical features of bilateral acute idiopathic maculopathy

PURPOSE: To describe the clinical course of bilateral acute idiopathic maculopathy (BAIM), and to analyze its pathophysiology. CASE: A 33-year-old Japanese woman presented with a sudden, severe, bilateral visual disturbance following a flu-like illness. She was examined by fluorescein angiography (FA), indocyanine green angiography (IA), scanning laser ophthalmoscopy (SLO), optical coherence tomography (OCT), and multifocal electroretinography (mfERG). OBSERVATIONS: A diagnosis of BAIM was made in this patient based on typical ophthalmoscopic features, which included a pathognomonic yellowish-white foveal lesion. FA demonstrated a breakdown of the outer blood-retinal barrier, with the size and location corresponding to the white lesion, and IA disclosed a choroidal circulatory disturbance. SLO demonstrated that the deep retinal and choroidal layers were disorganized, and OCT showed retinal edema. Electrophysiological dysfunction was detected by mfERGs. After steroid therapy, the patient's visual acuity recovered to normal. The pooling of fluorescein dye and the OCT-determined retinal edema were resolved. However, the physiological dysfunction detected by mfERGs remained. CONCLUSIONS: We conclude that the major abnormality in BAIM is an alteration of the retinal pigment epithelium causing severe edema.     

Polymorphisms in Complement Factor H and Hemicentin-1 genes in a Japanese population with dry-type age-related macular degeneration

PURPOSE: To determine whether polymorphisms in the Complement Factor H (CFH) gene and the Hemicentin-1 gene at the ARMD1 locus are associated with dry age-related macular degeneration (AMD) in Japanese patients. DESIGN: Clinically relevant laboratory investigation. METHODS: Eighty unrelated Japanese patients with dry AMD and 196 Japanese control patients were studied. Two exons of the CFH gene and four exons of the Hemicentin-1 gene were amplified by polymerase chain reaction and sequenced directly. RESULTS: For the CFH gene, the frequency of the previously reported Tyr402His variant was not significantly higher in the AMD group than in the control group (P = .31). In the Hemicentin-1 gene, three sequence alterations (Asp5088Val, IVS99-13C/T, and His5245Gln) were detected, and the originally reported Gln5346Arg was not detected. CONCLUSION: The CFH gene and Hemicentin-1 genes do not appear to be involved in a statistically significant fraction of dry AMD cases in the Japanese population.