Stage-specific expression of cancer-associated type 1 and type 2 chain polylactosamine antigens in the developing pancreas of human embryos.

Expression of type 1 and type 2 chain carbohydrate antigens during the course of morphogenesis of human embryonic pancreas was investigated using specific monoclonal antibodies and compared with the carbohydrate antigen profiles of human pancreatic cancers. The type 2 chain antigens, such as stage-specific embryonic antigen 1 (Le(x)) and I-antigens, appeared much earlier than the type 1 chain antigens; the epithelial cells of primitive foregut were Le(x)+I-antigen- in the embryos at Carnegie stages 16-23, while the pancreatic primordial cells, which had differentiated from the Le(x)+ gut epithelial cells, were Le(x)-I-antigen+ at Carnegie stages 22-23. The type 1 chain antigens, such as Le(a), Le(b), Le(c), and their sialylated derivatives, were not expressed in any cells at these stages and appeared much later in the pancreas of the 10-12-week embryos, when the primitive pancreatic ductal cells in the primordia exhibited an extensive budding of the daughter cells. At this stage, Le(a) appeared and was expressed strongly in the epithelial cells of primitive pancreatic ducts as well as in the daughter cells that were destined to differentiate into future centroacinar cells; Le(b) was localized in the daughter cells which were to become future acinar cells; and Le(c) was specifically expressed in the daughter cells which were to form future Langerhans islets. With regard to the sialylated derivatives of Le(a), expression of the 2-3 sialyl Le(a) antigen was limited to the epithelial cells of the primitive pancreatic ducts, while the 2-6 sialyl Le(a) antigen was strongly expressed in the future centroacinar cells, which had differentiated from the corresponding daughter cells. Among these antigens, the Le(a) and 2-3 sialyl Le(a) antigens showed the highest incidence in human pancreatic cancer tissues. These results indicate that the expression of these carbohydrate antigens in embryonic pancreas is differentiation dependent and cell lineage specific and that most human pancreatic cancer cells mimic the carbohydrate antigen profile of the epithelial cells of the primitive pancreatic ducts in human embryos.     

Carnitine and lipoate ameliorates lipofuscin accumulation and monoamine oxidase activity in aged rat heart

In this study we have focused on the levels of lipofuscin, monoamine oxidase and cholesterol phospholipid ratio in the heart muscle of young, middle aged and aged rats. In parallel, we have also investigated the levels of carnitine and lipoic acid during aging. We observed an increase in lipofuscin accumulation and monoamine oxidase activity in both middle aged and aged rats. Levels of both carnitine and lipoic acid decreased along with a decrease in cholesterol phospholipid ratio. These changes were normalized upon cosupplementation of carnitine and lipoic acid. Our results thus reveal that carnitine along with lipoic acid can be used as an effective supplement against free radical induced damage to the cardiac tissue.     

P-selectin-dependent macrophage migration into the tubulointerstitium in unilateral ureteral obstruction

BACKGROUND: Interstitial infiltration of macrophages (M?) is one of the main causal factors for the tubulointerstitial injury. However, precise mechanisms of M? infiltration into tubulointerstitium have not been fully explored. The purposes of this study were to assess the role of selectins in the acute infiltration of M? in rats with unilateral ureteral obstruction (UUO) and to evaluate the role of vasa recta, that is, whether they facilitate massive influx of M? into the interstitium by functioning as specialized vessels. METHODS: To evaluate the role of selectins in M? infiltration into tubulointerstitium, the expression of selectins and L-selectin ligands was examined by immunohistochemistry and immunoelectron microscopy. The functional role of P-selectin in vasa recta was studied by Stamper-Woodruff assay, in vivo p-M? migration assay and in vivo blocking experiments with the monoclonal antibody (mAb) ARP2-4. RESULTS: Selective expression of P-selectin was detected in vasa recta as early as one hour after UUO, and the expression increased thereafter for 96 hours. In contrast, endothelial expression of L-selectin ligands and E-selectin were not detectable. In the Stamper-Woodruff assay on kidney sections of rats with UUO, the adhesion of isolated rat peritoneal M? (p-M?) to vasa recta was significantly inhibited by the mAb ARP2-4 (P-selectin blocker; P < 0.01), but not by mAb ARE-5 (E-selectin blocker) or rLECIg (rat L-selectin chimeric protein). In the in vivo transfer experiments with fluorescein-labeled p-M? into rats 48 hours after UUO, labeled p-M? had accumulated around vasa recta at three minutes and had infiltrated predominantly into the outer medulla at 180 minutes. The number of labeled p-M? was reduced when the rats were pretreated with ARP2-4 (P < 0.01). Finally, ARP2-4 (10 mg/kg), injected 15 minutes before UUO, reduced the number of infiltrated M? (P < 0.01). CONCLUSION: The results suggest that vasa recta, which express P-selectin, contribute to massive infiltration of M? into the interstitium by functioning as specialized post-capillary venules.     

Diabetic alteration of cardiac vago-sympathetic modulation assessed with tone-entropy analysis

No clear evidence of diabetic alteration of cardiac sympatho-vagal balance has been reported to date. We assessed heart rate variability of diabetic patients with the tone-entropy analysis that has been published elsewhere (Oida et al. 1997). Tone reflects the cardiac vago-sympathetic balance and entropy the total autonomic efferent activity. Diabetic influence on tone and entropy was examined in two groups of patients (38-52 years and 60-69 year, total 106) stratified according to the occurrence of impaired glucose tolerance or diabetes mellitus. Ten healthy middle-aged volunteers were also examined as a reference. Electrocardiographic data were collected at rest for 10 min. Tone increased and entropy decreased significantly with severity of diabetic disorders. The alterations were depicted as a curvi-linear relation in tone-entropy space, which superimposed adequately on the standard tone-entropy values obtained in a pharmacological experiment. The results demonstrate that the vago-sympathetic balance is altered with diabetic disorders: vagal predominance is impaired significantly in proportion to a withdrawal of total autonomic efferent activity.     

Human erythroleukemia cell line (HEL) undergoes a drastic macrophage- like shift with TPA

We investigated the effect of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on the human erythroleukemia cell line, HEL, and found that TPA addition (10(-6)-10(-8) M) to HEL cell cultures induces morphological, functional, and biochemical changes in HEL cells that are characteristic for macrophage-like cells. Apart from the drastic changes in morphology, the cells greatly enhance their phagocytic ability and acquire receptors for binding and degradation of chemically modified lipoproteins. At the biochemical level, a newly synthesized 85K glycoprotein is observed, and the cells are unresponsive to inducers of globin synthesis. Comparative observations with K562 cells indicate that TPA inhibits, as in HEL cells, spontaneous and induced globin synthesis, but induces minimal macrophage-like properties in these cells. The results with HEL cells are interpreted to indicate that TPA uncovers a latent monocyte-like phenotype in these cells.     

Oxidative stress-mediated macromolecular damage and dwindle in antioxidant status in aged rat brain regions: role of L-carnitine and DL-alpha-lipoic acid

BACKGROUND: The free radical theory of aging has significant relevance in a number of age-related neurological disorders. Too many free radicals create cellular pollution that shuts down energy levels. They have also been implicated in the loss of physiological functioning associated with the aging of post mitotic cells such as the brain. The activities of enzymatic antioxidative defenses decrease in rat brain may be possible causes of age-associated increase in oxidative damage to macromolecules. METHODS: We determined whether DL-alpha-lipoic acid (100 mg/kg body weight/day), and L-carnitine (300 mg/kg body weight/day) together when administered for 30 days declines the rate of oxidative stress-mediated macromolecular damages such as lipid peroxidation (LPO), protein carbonyl (PCO) and DNA protein cross-links in different anatomic regions (cortex, striatum and hippocampus). The activities of antioxidant enzymes in programmed aging were evaluated in the cortex, striatum and hippocampus of young and aged rat brain regions. RESULTS: Aged rats elicited a significant decline in the antioxidant status and increase in LPO, PCO and DNA protein cross-links as compared to young rats in all the 3 brain regions. The increase in LPO, PCO and DNA protein cross-links were (35.8%, 35.6%, 43.5%) in cortex, (32.5%, 40.3%, 29.8%) in striatum and (62.7%, 42.4%, 34.9%) in hippocampus, respectively, in aged rats as compared to young rats. Co-supplementation of carnitine and lipoic acid was found to be effective in reducing brain regional LPO, PCO and DNA protein cross-links and in increasing the activities of enzymatic antioxidants in aged rats to near normalcy. CONCLUSION: The combination of l-carnitine and lipoic acid overcame the oxidative stress induced rate of lipid peroxidation, protein carbonyl formation, accumulation of DNA protein cross-links and deficits in antioxidant enzyme activities in various brain regions of aged rats.     

Immunological aspects of rat models of HTLV type 1-infected T lymphoproliferative disease

The level of host immune responses against human T cell leukemia virus type 1 (HTLV-1) varies among HTLV-1-infected individuals. In the present study, we investigate the role of host immunity on HTLV-1 leukemogenesis in vivo by using animal models. At first, we examined the effect of the routes of HTLV-1 transmission on the host anti-HTLV-1 immune responses. When immune competent adult rats were inoculated with HTLV-1-infected cells, the orally infected rats were persistently infected with HTLV-1 without humoral and cellular immune responses against HTLV-1, whereas all intravenously or intraperitoneally inoculated rats showed significant levels of immune responses. Next, we examined in vivo tumorigenicity of HTLV-1-immortalized cells in the absence of T cell immunity, by using athymic F344/N Jcl-rnu/rnu (nu/nu) rats. When inoculated into nu/nu rats, not all but some HTLV-1-immortalized rat cell lines including syngeneic FPM1-V1AX could grow and form T cell lymphoma in vivo. This syngeneic lymphoma formation was inhibited by adoptively transferred immune T cells. Furthermore, immunocompetent rats allowed in vivo growth of HTLV-1-infected lymphoma, when treated with antibodies that block costimulatory signals for T cell activation. These observations indicated that (1) host anti-HTLV-1 immunity can be affected by the conditions of the primary infection, (2) under the low pressure of anti-HTLV-1 immunity, some HTLV-1-infected cell clones grow in vivo, and (3) T cell immunity is required for in vivo surveillance against these HTLV-1-infected cell clones.     

Modulation of age-associated oxidative DNA damage in rat brain cerebral cortex, striatum and hippocampus by L-carnitine

The free radical theory of cell aging may have significant relevance in the pathogenesis of a number of age-related neurological disorders. A large body of experimental evidence supports the existence of a relationship between genomic instability, DNA damage and aging. The age-associated accumulation of oxidative DNA damage is well documented in central nervous system. The decline of mitochondrial respiratory function and loss of normal cellular homeostasis as consequences of excessive accumulation of endogenous oxidative damages to DNA have long been indicated in the aging process. In the present study, age-associated alterations in the content of DNA and accumulation of oxidative DNA damage products such as 8-OHdG and DNA protein cross-links are mainly focused. In parallel, we have also investigated the salubrious effect of l-carnitine against oxidative DNA damage as it possesses energy and antioxidant improving properties. Our results thus reveal that L-carnitine has inhibiting effect on the accumulation of age-related oxidative DNA damage in various brain regions, viz. cerebral cortex, striatum and hippocampus.     

An efficient tool for surveying CRF01_AE HIV type 1 resistance in Thailand to combined stavudine-lamivudine-nevirapine treatment: mutagenically separated PCR targeting M184I/V

Under programs organized by the government of Thailand, HIV-1-infected patients have been treated since 2002 with several regimens, including a tablet known as GPOvir, which contains lamivudine, stavudine, and nevirapine. The aim of this study was to establish an effective assay, based on mutagenically separated PCR (MS-PCR), with the goal of surveying GPOvir-resistant HIV-1 cases. To determine the target mutation point for the assay, we analyzed the patterns of acquired drug resistance in plasma samples from GPOvir-failed cases. Of 428 HIV-1-infected individuals treated with GPOvir at Lampang Hospital in northern Thailand from 2002 to 2004, 66 had detectable viral loads after 3 months of treatment. The HIV-1 sequences of these 66 GPOvir-failed cases and 55 pre-GPOvir baseline samples were analyzed. The most prevalent drug resistance mutation among the samples was the lamivudine resistance M184I/V mutation. Based on this finding, we developed a new MS-PCR assay to detect the M184I/V mutation, and evaluated the assay performance for detecting GPOvir-resistant CRF01_AE cases. Comparing the results of M184I/V MS-PCR and sequence analyses, we found a concordance rate of 95% and an overall sensitivity of the M184I/V MS-PCR for detecting GPOvir-resistant cases of 79%. Considering the relatively low price of the assay, approximately $12.50 per sample, M184I/V MS-PCR may be a candidate for monitoring a large number of GPOvir-treated patients, particularly in developing nations.