Enhancing effects of oral adjuvants on anti-HBs responses induced by hepatitis B vaccine.

Hepatitis B (HB) vaccine is very promising for the prevention of HB infection. There exist, however, some non-responders to current vaccination trials. In this study, taurine, parotin and lithium were selected as adjuvants which can be administered orally. The mechanisms of these three materials as adjuvants and their effects on HB vaccine were investigated in mice. For instance, taurine induced polyclonal antibody production and exhibited adjuvant activity. Although taurine did not have any activity on the proliferation of thymocytes nor stimulate IL-2 production, taurine did induce IL-1 production by macrophages. It was considered that taurine-induced IL-1 would play an essential role in the proliferation and differentiation of B cells. Parotin also induced polyclonal antibody production and exhibited adjuvant activity. These effects of parotin were not affected even if macrophages or T cells were depleted, and parotin itself had an IL-1-like activity. Therefore, it was considered that parotin acted directly on B cells by its IL-1-like activity and mitogenic activity, resulting in the proliferation and differentiation of B cells. Lithium induced neither polyclonal antibody production, nor IL-1 or IL-2 production. However, when given with an antigen, lithium activated the humoral immune system, resulting in the augmentation of antibody production. Oral administration of taurine, parotin and lithium were capable of restoring antibody responses to HB surface antigen (HBsAg) in HBsAg-nonresponder mice. Furthermore, taurine, parotin and lithium enhanced the adjuvant effects of aluminium contained in the present HB vaccine. These observations indicate that use of these oral adjuvants may open new perspectives in the field of human HB vaccination.     

Molecular biology techniques as clinical laboratory tests

The quality of results obtained with molecular biology techniques depends on the control of preanalytical and analytical error associated with such techniques. Preanalytical error can be introduced during the isolation of DNA and RNA. The type of detergent used in cell lysis can affect the amplification of DNA by techniques such as the polymerase chain reaction(PCR). Ribonuclease(RNase) contamination is a serious problem in the isolation of undegraded RNA, and, thus, this enzyme should be inhibited. Anticoagulants used for blood collection can affect the quality of results with molecular biology techniques. The control of contamination from the working environment is essential to the minimization of preanalytical, analytical and postanalytical error. Molecular biology techniques for a wide range of clinical laboratory tests have been established in hospitals such as clinical laboratory tests for infections, molecular diagnoses of leukemia and aberrant genes in metabolism.     

Immunohistochemistry and angiography in adenomatous hyperplasia and small hepatocellular carcinomas

The sinusoidal structure and blood supply of 38 liver nodules less than 2 cm In diameter were Investigated. There were 18 cases of adenomatous hyperplasia (AH) and 20 cases of hepatocetlular carcinoma (HCC). Growth pattern, encapsulation and vascularity were examined, and Immunohistochemistry performed for factor VIII related antigen (factor VIII), type IV collagen (collagen IV), lamlnln and CD68. There were significant differences between AH and small HCC, except for the expression of CD68. There were differences In tumor size, vasculartty and the components of the basement membrane between AH and small, well differentiated HCC. The cases of AH were supplied by the portal system and maintained the sinusoidal structure, but small well-differentiated HCC were supplied by a mixture of portal and arterial vessels. In spite of their small size, moderately and poorly differentiated HCC had capillary and were supplied by branches of the hepatic artery.     

Association of stress proteins with autoantigens: a possible mechanism for triggering autoimmunity?

Patterns of autoantibody production are diagnostic of many autoimmune disorders; the recent observation of additional autospecificities towards stress-induced proteins may also provide insight into the mechanisms by which such responses arise. Grp78 (also known as BiP) is a target of autoaggressive B and T cell responses in our murine model of anti-Ro (SS-A) autoimmunity and also in rheumatoid arthritis. In this report we demonstrate reciprocal intermolecular spreading occurs between Ro52 and Grp78 in immunized mice, reflecting physiological association of these molecules in vivo. Moreover, we provide direct biochemical evidence that Grp78 associates with the clinically relevant autoantigen, Ro52 (SS-A). Due to the discrete compartmentalization of Ro52 (nucleocytoplasmic) and Grp78 (endoplasmic reticulum; ER) we propose that association of these molecules occurs either in apoptotic cells, where they have been demonstrated indirectly to co-localize in discrete apoptotic bodies, or in B cells themselves where both Ro52 and Grp78 are known to bind to immunoglobulin heavy chains. Tagging of molecules by association with Grp78 may facilitate receptor mediated phagocytotsis of the complex; we show evidence that exogenous Grp78 can associate with cell surface receptors on a subpopulation of murine splenocytes. Given the likelihood that Grp78 will associate with viral glycoproteins in the ER it is possible that it may become a bystander target of the spreading antiviral immune response. Thus, we propose a model whereby immunity elicited towards Grp78 leads to the selection of responses towards the Ro polypeptides and the subsequent cascade of responses observed in human disease.     

Cytokeratin subunits of inclusion bodies in rhabdoid cells: immunohistochemical and clinicopathological study of malignant rhabdoid tumor and epithelioid sarcoma

Extrarenal malignant rhabdoid tumor (MRT), which is recognized as being histologically similar to renal MRT, is characterized by the presence of "rhabdoid cell" (RC) and a highly aggressive biological behavior. Recently it has been proposed that "proximal variant" of epithelioid sarcoma (ES), whose morphology is similar to that of MRT, actually has a more aggressive clinical course than classical type ES. Detailed immunohistochemical analysis of cytokeratin (CK) subunits was performed in 3 cases of extrarenal MRT, 3 cases of renal MRT, and 11 cases of ES comprising 2 "proximal variants" and 9 classical types. Renal and extrarenal MRTs showed positive immunoreactivity for both CK8 and CK18. Classical type ESs were diffusely positive, not only for CK8 and CK18, but also for other cytokeratin subunits including CK4, 6, 10, 13, 16, 17, and "high-molecular-weight" CKs (CK1, 5, 10, and 14). On the other hand, proximal ES revealed limited immunohistochemical reactivity for cytokeratins, compared with classical ES. In conclusion, the inclusion bodies of RCs show immunoreactivity confined to CK8, CK18, and vimentin. Furthermore, ES has additional CK expressions, while proximal ES possesses characteristics intermediate between those of classical ES and those of external MRT.     

Vinyl polymerization. 120. Mutual copolymerization of p-substituted styrenes through radical mechanism

The radical mutual copolymerization of p-substituted styrenes, such as p-methoxy-, p-chloro-, p-bromo-, p-cyanostyrene, and styrene was carried out with one another at 30°C. in the dark. As initiator, azobisisobutyronitrile was used. The plots of the copolymerization rates against HAMMETT 's σ values showed no linear relationships and the concave curves were obtained therefrom. The relative reactivities of p-substituted styrenes with a definite p-substituted polystyryl radical, which were shown by the reciprocal of monomer reactivity ratio r₁, were plotted against σ values and concave curves were also obtained. The relative reactivities of p-substituted polystyryl radicals with p-substituted styrene were calculated from the ratios r₂ and the propagation rate constants in homopolymerization. the plots of them against σ values gave straight lines with different ρ values, according to the polarity of substituents. These results suggest that polar structures in transition state affected markedly the copolymerization rates. The effect of substituents on resonance stabilization was also quantitatively estimated.     

Differences in the abilities of individual fingers during the performance of fast,repetitive tapping movements

Using 12 healthy male subjects, the dynamic motor ability of individual fingers was investigated under four different finger tapping conditions. These were: maximum speed tapping with one finger (single-finger tapping), alternate movement of two fingers (double-finger tapping), double-finger tapping in an unsupported condition, and submaximum constant speed tapping with one finger in a passive manner. Key-contact forces for all fingers and the movement velocity of the tapping finger were monitored. With the exception of the unsupported condition, non-tapping fingers were maintained in contact with designated keys during the tapping tasks. It was found that the index finger attained the fastest cadence and greatest movement velocity, followed by the middle, little and ring fingers, respectively. Subjective assessment of rank order of "difficulty" of tapping by the subjects was highly correlated with tapping cadence. Thus dynamic motor function, as indicated by rapid, repetitive movement, differs among the individual fingers. Parallel changes were observed in the key-contact force of the neighboring non-tapping fingers during tapping. The range of the non-tapping finger forces was largest during tapping by the ring finger. A similar trend was found for passive tapping, during which the magnitude of key-contact force was less than one-third of that observed during active tapping. The lower cadence achieved by the ring finger may be attributed more to a lack of independence at the level of voluntary neuromuscular control, than to innate mechanical interaction with the other fingers. Tapping cadence of each finger was lower for the double-finger mode than for the single-finger mode. The magnitude of the observed decrease in cadence during double-finger tapping was, on the other hand, strongly dependent on finger-combination. The decrease was smallest for the index-middle finger-combination, and greatest for the ring-little finger-combination. Compatibilities with other fingers can play an essential role in the dynamic motor function of individual fingers. During the unsupported task, in which interactions were diminished by allowing all fingers to move freely, tapping cadence increased markedly. Therefore, the lower cadences observed in specific finger-combinations may be partly attributed to anatomical and neural interdigit interactions.     

Proliferation,apoptosis, and intratumoral vascularity in multiple myeloma: correlation with the clinical stage and cytological grade

AIMS: Abnormalities involving proliferation, apoptosis, and angiogenesis are important in tumorigenesis. The purpose of this study was to examine these three biological processes, and their relation with the clinical stage and cytological grade in multiple myeloma (MM). METHODS: Fifty four newly diagnosed patients with MM were studied by immunohistochemistry using bone marrow clot sections. Proliferation and apoptosis were evaluated for the proportion of MM cells (indicated by morphology and CD138 reactivity) positive for the Ki67 antigen and single stranded DNA (ssDNA), respectively. Angiogenesis was evaluated by measuring the intratumoral microvessel density (IMVD) and by assessing the immunoreactivity of vascular endothelial growth factor (VEGF). RESULTS: There were 30 men and 24 women (median age, 65 years; range, 37-84). At initial presentation, 15 (28%) were in Durie stage I, 15 (28%) in stage II, and 24 (44%) in stage III. Advanced clinical stage correlated with high cytological grade (p < 0.03). The medians for Ki67, ssDNA, and IMVD were 4.4% (range, 0-15%), 0.2% (range, 0-2.8%), and 15.5 (range, 0-63), respectively. Among these three continuous parameters, the only significant correlation was that between Ki67 and IMVD (p < 0.0001). Both Ki67 and IMVD also correlated with the clinical stage, cytological grade, and VEGF positivity (p <0.05). No correlation was found between ssDNA and all of the other parameters. CONCLUSIONS: These data suggest that proliferation is associated with angiogenesis in MM. Furthermore, proliferation and angiogenesis, but not apoptosis, may be important in disease progression. Lastly, increased production of VEGF may be one of the contributing factors to the increase in intratumoral vascularity seen in advanced MM.