Regeneration of beta cells in the native pancreata after syngeneic and allogeneic pancreas transplantations in spontaneously type 2 diabetic Torii rats.

BACKGROUND: We previously demonstrated that syngeneic pancreas transplantation has a potential to reverse diabetes even in a rat model of type 2 diabetes mellitus, namely Spontaneously Diabetic Torii (SDT; RT1a). The onset of diabetes was significantly delayed in the pancreas transplant recipients. We speculated that perfect diabetic control achieved by pancreas transplantation showed a beneficial effect on the native pancreata & recipients. MATERIALS AND METHODS: Twenty-five-week-old diabetic SDT rats were divided into 3 groups: untreated controls and syngeneic and allogeneic transplant recipients. We transplanted pancreaticoduodenal grafts from nondiabetic 10-week-old SDT rats and from 10-week-old allogeneic Dark Agouti (DA; RT1a) rats using daily administration of FK506. RESULTS: Untreated SDT rats showed disappearance of pancreatic and duodenal homeobox-1 (PDX-1) expression in the pancreas and a marked decrease in beta-cell mass. Among syngeneic and allogeneic pancreas transplant recipients, islet-like cell clusters were found in the native pancreata. The beta-cell mass at 40 weeks of age was significantly increased in the native pancreata of recipients compared with age-matched controls. Interestingly, we observed the reexpression of PDX-1 in the nuclei of islet-like cell clusters. CONCLUSIONS: Our results indicated the benefits of avoiding glucose toxicity by pancreas transplantation which induced PDX-1 expression in the native pancreata of type 2 diabetic recipients, resulting in regeneration of beta cells in the native pancreata.

Adenoviral-mediated overexpression of either membrane-bound human FasL or human decoy Fas can prolong pig islet xenograft survival in a rat transplant model.

The success of pancreatic islet transplantation is limited because of the severe shortage of allogeneic pancreas donors. Accordingly, pig islets are considered to be an attractive, promising alternative. However, cell-mediated immunity, especially CD8+ cytotoxic T lymphocyte (CTL)-mediated cytotoxicity, remains a formidable barrier to prevent long-term islet survival in xenograft recipients. Therefore, it is particularly important to explore methods to specifically prevent cell-mediated immunity against pig islets. Our group previously demonstrated that the overexpression of either membrane-bound human FasL or human decoy Fas antigen in pig endothelial cells prevented CTL xenocytotoxicity. In this study, we assessed the cytoprotective effects of adenoviral-mediated overexpression of either membrane-bound human FasL or human decoy Fas antigen in pig islets to inhibit CTL xenocytotoxicity. The CTL-mediated killing of pig islets infected with an adenoviral vector carrying either membrane-bound human FasL or human decoy Fas was significantly reduced compares with that of control pig islets transfected with adenoviral vector encoding enhanced green fluorescent protein (EGFP). Moreover, we transfected pig islets with these molecules to confirm their cytoprotective effects in in vivo studies. The significant long-term survival of pig islets expressing these molecules was elicited through days 3 to 5 posttransplantation. Thus, these results demonstrated that the remodeling of either death receptor or death ligand on pig islets by adenoviral gene transfer prevented innate cellular immunity against xeno-islet grafts facilitating long-term xenograft survival.

Adenoviral-mediated overexpression of membrane-bound human FasL and human decoy Fas protect pig islets against human CD8+ CTL-mediated cytotoxicity.

Pig islets are considered to be most suitable source of islets for xenotransplantation into patients with type 1 diabetes mellitus. However, cellular rejection, especially CD8+ CTL-mediated cytotoxicity, remains a formidable barrier preventing long-term xenograft survival. Our previous study demonstrated that human CD8+ CTLs were highly detrimental to xenograft cells and that this strong cytotoxicity of human CTLs was mediated mainly by the Fas/FasL apoptotic pathway. Furthermore, we exploited novel methods for inhibiting human CD8+ CTL-mediated xenocytotoxicity with overexpression of membrane-bound human FasL and human decoy Fas antigen in xenografted cells. In the present study, we assessed the cytoprotective effects of these novel inhibitory molecules overexpressed by an adenoviral-mediated system in pig islets. Isolated pig islets were transfected with adenovirus vector encoding either human decoy Fas or membrane-bound human FasL genes. Thirty percent to 60% of transfected pig islets expressed these molecules producing 60% to 88% suppression of CTL killing compared with parental pig islets. These data indicated that pig islet grafts isolated from transgenic pigs with either membrane-bound human FasL or human decoy Fas antigen genes may control the innate cellular response to xenografts, and creating a window of opportunity to facilitate xenograft survival.

Sequence-based structural features between Kvlqt1 and Tapa1 on mouse chromosome 7F4/F5 corresponding to the Beckwith-Wiedemann syndrome region on human 11p15.5: long-stretches of unusually well conserved intronic sequences of kvlqt1 between mouse and human.

Mouse chromosome 7F4/F5 is a syntenic locus of human 11p15.5 in which many imprinted genes are clustered. Transmission of aberrant human 11p15.5 or duplicated 11p causes Beckwith-Wiedemann syndrome (BWS) depending on which parent the chromosome is derived from. To analyze a syntenic mouse locus corresponding to human 11p15.5, mouse BAC contigs were constructed between Nap2 and Tapa1, in which 390 kb was sequenced between Kvlqt1 and Tapa1. An unexpected finding was that of highly conserved intronic sequences of Kvlqt1 between mouse and human, and their homologies came up to at least 160 kb because the length of this gene extended to 350 kb, suggesting the possibility of some functional constraint due to transcriptional and/or post-transcriptional regulation of this region. Many expressed sequence tags (ESTs) were mapped on this locus. Three genes, Lit1 (Kvlqt1-AS), Mtr1 and Tssc4, were identified and characterized. Lit1 is an antisense-transcript of Kvlqt1 and paternally expressed and maternally methylated throughout the developmental stage. The position where Lit1 exists corresponded to a highly conserved region between mouse and human. This transcript extends at least 60 kb from downstream to upstream of exon 10 in Kvlqt1. Tssc4 and Mtr1 carried putative open reading frames but neither was imprinted. Further characterization of this locus based on the sequence comparison between mouse and human will contribute valuable information towards resolving the mechanism of the occurrence of BWS and the associated childhood tumor.

[Experience with the Groshong port in a home TPN patient--a case report].

We experienced a home TPN patient using Groshong port. A 45-year-old woman, recurrent pancreatic head cancer, was treated with home TPN. The Groshong port was inserted via external jugular vein and stayed at home for 8 months with home TPN. Lipid emulsion was administered twice a week. After administration of lipid emulsion, 20 ml of saline was injected and catheter lock was performed with only saline. Heparin was not used for catheter lock. After 8 months' home TPN, patient died. We could not find any precipitate in the catheter and reservoir. A unique feature of Groshong port is a pressure-sensitive two-way valve at the intravascular end. The valve of this port opens outward during fluid infusion or bolus injection. During blood withdrawal, the valve opens inward and then closed after the procedure. Because the valve is designed to remain closed between -7 and 80 mmHg, it will not open spontaneously under normal central venous pressure conditions. This valve eliminates the need for heparin flusing. Catheter obstruction during prolonged TPN is a serious complication. Compound of heparin, lipid emulsion and electrolytes, especially calcium is now becoming one of the most important causes of catheter obstruction and catheter removal. We used the Groshong port to prevent catheter obstruction. After 8 months' use of this port, no precipitate was found in this port. We conclude that the Groshong port is useful for home TPN catheter because it can prevent catheter obstruction due to heparin-lipid-electrolytes compound.

[Home parenteral nutrition for terminal cancer patients--when should we tell the truth about their disease and return them to their home with HPN].

Home parenteral nutrition (HPN) is a significant way to enable terminally ill cancer patients to live with high quality of life at home. However, the most important condition to perform HPN is the patients' desire to return to their home. For this purpose, we should consider in informing them of the truth of their disease. We had three terminal cancer patients who returned to their home with HPN. Cas 1: 52-year-old woman, recurrent stomach cancer. She did not know her cancer. She stayed at home for 45 days with HPN. Case 2: 46-year-old woman, recurrent stomach cancer. She did not know her recurrence of cancer. She stayed at home for only 15 days with HPN. Case 3: 45-year-old woman, recurrent pancreatic head cancer. She knew her disease and her life expectancy. She had 6-year-old daughter and desired to return home. She stayed at home with HPN for 8 months. As we told Case 3 truth of her disease when she was diagnosed as recurrence of cancer, she lived at home with HPN for 8 months. From the experience of three cases, we may concluded that, the sooner the patients are informed the truth about their disease, the longer they can stay at home with HPN and spend their limited time with high QOL.

Incidence of human T lymphotropic virus type I seroconversion after age 40 among Japanese residents in an area where the virus is endemic.

To clarify how seroconversion after age 40 years contributes to the seroprevalence of human T lymphotropic virus type I (HTLV-I), Japanese residents were studied in an area of Kyushu in which HTLV-I is endemic. Cross-sectional data showed higher seroprevalences among subjects > or = 40 than in those < 40 years old (27.0% vs. 7.9%). The prevalence was also greater in women than men (25.5% vs. 17.6%). Six hundred subjects > or = 40 years old were tested during 1976-1993. Among 8 who seroconverted, 5 had an HTLV-seropositive spouse and 2 seroconverted after blood transfusions. The incidences per 1000 person-years of seroconversion in age groups 40-49, 50-59, and > or = 60 years were, respectively, 0.0, 0.0, and 3.3 (range, 1.38-5.13) in men and 0.0, 5.3 (range, 0.11-7.99), and 1.4 (range, 0.00-1.60) in women. The seroconversion incidence was insufficient to account for the rise of seroprevalence in those > or = 40 years old, especially among women. These results suggest that the high rate and sex discordance in HTLV-I seroprevalence after age 40 may depend mainly on the birth cohort effect for vertical or horizontal transmission.

Seroepidemiology of adult T-cell leukemia virus infection and analysis of sero-positive cases in Taiwan.

For a seroepidemiologic study of adult T-cell leukemia virus (ATLV) infection in Taiwan, the gelatin particle agglutination technique and the indirect immunofluorescence method were used for anti-ATLV titration. Sporadic sero-positive cases were found all over the Taiwan districts except among the aborigines (0/947). Sero-positive rates ranged from 0 to 5.6% (except ATL family) and a total of 48 cases were found in 3682 Han-Chinese. Among them 9 cases were newly found in family surveys, and 39 cases were observed in random samples. As an average positive rate was 1.0%, by calculation about 80,000 sero-positive cases are supposed to be present in Taiwan. A most remarkable feature of the sero-positive cases was the high rate in couples. Various patterns of sero-positive cases existed in pedigrees. Anti-ATLV positive sera of Chinese living in Taiwan and Japanese were compared by immunoprecipitation and there was no difference between them. The possible infection route from Japan to Taiwan is discussed.

HTLV-I carriers among migrants from an ATL-endemic area to ATL non-endemic metropolitan areas in Japan.

The prevalence of antibodies against HTLV-I among Kyushu natives aged 16 to 39 years who moved from Nagasaki and Kagoshima prefectures to Aichi prefecture (a non-endemic area for ATL) was compared by their cities or counties of birth. The positive rate of anti-HTLV-I antibody was 2.4% (II/400) among Nagasaki natives, 6.4% (20/312) among Kagoshima natives and 4.0% (31/772) for both combined. There was a slight difference in the positive rate of anti-HTLV-I antibody between Kyushu natives from cities (3.3%) and from counties (4.5%). In county areas, the prevalence of anti-HTLV-I antibodies among migrants from areas of relatively higher mortality for malignant lymphomas (7.5%) was significantly higher (p less than 0.01) than among persons from lower mortality areas (1.9%). Most "positive" persons had moved from Kyushu to Aichi prefecture between the ages of 15 and 18 years. The results of the present study suggest that: there is a considerable number of HTLV-I carriers among Kyushu natives who have settled in ATL non-endemic areas, especially among those born in regions of Kyushu district which have a high mortality rate for malignant lymphomas; and that Kyushu natives who had settled in metropolitan areas might have been exposed to HTLV-I during childhood in their birthplace.

Seroepidemiology of the human retrovirus (HTLV/ATLV) in Okinawa where adult T-cell leukemia is highly endemic.

A survey of carriers of antibodies to HTLV-specific antigens was made in 11 islands among 60 Okinawan islands to examine the regional variation in the prevalence of HTLV carriers. The overall seropositive rate was 21% among 7,545 adults aged 40 years or more. The incidence of virus carriers was low in only one of the 11 islands (1-6%), while it was 14-31% in the other 10 islands.

Antibody to HTLV-I in indigenous inhabitants of the Andes and Amazon regions in Colombia.

To explore the HTLV-I-carrying groups among the indigenous inhabitants in South America, a sero-epidemiological study on HTLV-I focusing on hinterland villages isolated from others in the Andes and Amazon regions was conducted. Five (2.9%) out of 171 subjects showed positive for HTLV-I antibody in the gelatin particle agglutination (PA) test. Two out of 5 positives with high antibody titer (greater than or equal to x 1024) in the PA test also showed a positive immunofluorescence (IF) test and anti-HTLV-I-specific protein products, p19, p24, p28, gp46, and p53 in sera by the Western blotting (WB) test. One of three negatives in the IF test showed positive antibodies to p19 and p24 by the WB test. Finally, two were confirmed as HTLV-I carriers and one was suspected of being a carrier. All three are Paez Indians from the central Andes; 53- and 34-year-old women and a 35-year-old man. The results show that HTLV-I carriers exist among isolated indigenous people in South America.

Interleukin-6 levels in colorectal cancer tissues.

Interleukin-6 (IL-6) levels in cancer tissues of 32 patients with colorectal cancer were measured and analyzed in relation to the clinico-pathologic findings. The IL-6 level in cancer tissues was 81.1 +/- 118.0 pg/mg protein, which was significantly higher than the 11.2 +/- 20.8 pg/mg protein in normal colorectal tissues (p = 0.028). IL-6 levels in cancer tissues showed no correlation with the incidence of venous invasion, lymphatic invasion, lymph node involvement, or liver metastasis. The IL-6 level in cancer tissue tended to decrease as the cancer penetration extended. Our results suggest that IL-6 may play a role in human colorectal cancer growth.

Seroepidemiology of adult T-cell leukemia virus in Taiwan.

A survey of carriers of adult T-cell leukemia virus (ATLV) detected as anti-ATLV associated antigens was made in Taiwan. Among 2545 adults aged 30 years or more examined, seropositive donors amounted to 0.9% in the Han Chinese but none in the aborigines.

A new agglutination test for serum antibodies to adult T-cell leukemia virus.

A new gelatin particle agglutination test was developed for assay of natural antibodies to adult T-cell leukemia virus (ATLV/HTLV-I). Partially purified viral antigen from culture fluid of a virus-producer cell line was coated on artificial gelatin particle carriers. A high correlation was observed between the titers of antibodies determined by the agglutination test and by indirect immunofluorescence assay. The agglutination test is simple, sensitive and specific, and should be useful for mass screening of human sera for viral antibodies.

Serologic and epidemiologic studies on adult T-cell leukemia (ATL): special reference to a comparison between patients with and without antibody to ATL-associated antigen.

A case of adult T-cell leukemia (ATL) in a 23-yr-old Japanese man without immunoglobulin (Ig)G antibody to ATL-associated antigen (ATLA) (anti-ATLA) is presented. Comparative studies revealed that there were no differences between the present case and the anti-ATLA-positive ATL cases, except for the onset of the former at a younger age. Serologic and epidemiologic studies were performed to determine whether in this case the disease was still dependent on ATLA. As serum IgE and IgA had been persistently elevated, the serum was examined for these antibodies to ATLA. Furthermore, after the patient's serum was treated with glycine buffer to dissociate possibly existing immune complexes, it was assayed for antibodies (IgG, IgE and IgA) to ATLA. However, no antibody activity could be demonstrated. Sero-epidemiologic studies revealed that there were no anti-ATLA-positive healthy adults (ATLA-HA) in either his family or among the residents of his birthplace, whereas some ATLA-HA have been found among the residents of the birthplaces of other ATL patients. Although further investigation is necessary, it seems reasonable for the present to consider that a minor group of ATL patients independent of ATL virus may exist.

Epidemiological analysis of the distribution of antibody to adult T-cell leukemia-virus-associated antigen: possible horizontal transmission of adult T-cell leukemia virus.

About 20% of healthy inhabitants in the endemic area of adult T-cell leukemia/lymphoma (ATL) in Nagasaki Prefecture showed a positive antibody to adult T-cell leukemia virus (ATLV)-associated antigen (ATLA), while none of the inhabitants in a non-endemic area in Aichi Prefecture showed a positive anti-ATLA. The age-specific positive rate of anti-ATLA in the endemic area increased with age, especially in females. The positive rate of anti-ATLA among inhabitants older than 40 years was 21% in males and 36% in females. From epidemiological analyses of the mode of anti-ATLA reactivity among married couples and some family members and relatives with positive and negative anti-ATLA, two routes of transmission of ATLV were suggested. One route is from parents to children, either vertically or horizontally, and the other route is horizontal transmission between spouses, especially from husband to wife. The presence of some vectors of ATLV in the endemic area of ATL is discussed.

Geographical distribution of subjects seropositive for human T-cell leukemia virus type 1 in Papua New Guinea.

Of 1471 sera collected from 1986 to 1989 in Papua New Guinea (PNG), 2.2% were found to be positive for anti-HTLV-1 antibody by successive particle agglutination and immunofluorescence tests. The seropositive rate varied in different provinces and was higher in the coastal areas of the main island and in neighboring small islands than in the highlands of PNG. The frequency of HTLV-1 infection of children was higher, but the age-dependent increase in antibody positivity, generally observed in other HTLV-1 endemic areas of the world, was not clear in PNG. No difference was observed in antibody prevalence in males and females in this study.

Sporadic carriers of human T-lymphotropic virus type I in northern Egypt.

Sera from 3,158 individuals living in northern Egypt were tested for the presence of antibodies against human T-lymphotropic virus type I (HTLV-I) by the newly developed particle agglutination (PA) test. Ten sera gave a positive reaction in the PA test. Eight of these sera were examined further by Western blotting and all of them gave several bands corresponding to HTLV-I structural proteins. Two of the 8 sera gave positive results in the indirect immunofluorescence test. The results indicate the presence of HTLV-I carriers in this area, although at very low incidence (0.063%).

Incidence of adult T-cell leukemia-lymphoma and its familial clustering.

The yearly incidence of ATLL in the Uwajima district is 6.6 patients per 100,000 inhabitants aged over 40. The yearly morbidity rate from ATLL of persons in this district who are positive for HTLV-antibody and older than 40 is 1 patient per 1,631. Familial occurrence was observed in 9/38 families available for pedigree analyses. Even in the endemic area, the existence of positive HTLV antibody is remarkably high in ATLL families, suggesting that HTLV has been transmitted from generation to generation mainly within these particular families.