Hoek's formula, a cystatin C-based prediction formula for determining the glomerular filtration rate, is the most effective method for original adjusting the dosage of vancomycin.

OBJECTIVE: Some formulas using the serum cystatin C level to estimate the GFR have recently been reported. However, there has been no report of a serum cystatin C-based formula for adjusting the dosage of the drugs cleared by the kidney. In this study, we compared the predictive performance of the serum vancomycin trough concentration predicted using serum cystatin C-based formulas. METHOD: The data were collected from 158 hospitalized patients. Five formulas have been published to predict the GFR using serum cystatin C. The cystatin C-based formulas were divided into two groups, formulas with or without anthropometric data. We predicted the serum vancomycin trough concentrations using VCM-TDM S_edition ver. 1.00 software. RESULTS: In formulas with anthropometric data, the mean absolute error (MAE) using Hoek's formula was 2.38, the MAE using Grubb's 1 formula was 4.13, the MAE using Sjöström's formula was 2.90, and the MAE using Cockcroft and Gault formula based on creatinine was 4.42. On the other hand, in formulas without an anthropometric data group, the MAE using Larsson's formula was 3.07, and the MAE using Grubb's 2 formula was 3.63. CONCLUSION: These results suggested that Hoek's formula is the most useful formula for determining the initial dosage settings for vancomycin.

Galectin-1 regulates initial axonal growth in peripheral nerves after axotomy.

The signals that prompt the axons to send out processes in peripheral nerves after axotomy are not well understood. Here, we report that galectin-1 can play an important role in this initial stage. We developed an in vitro nerve regeneration model that allows us to monitor the initial axon and support cell outgrowth from the proximal nerve stump, which is comparable to the initial stages of nerve repair. We isolated a factor secreted from COS1 cells that enhanced axonal regeneration, and we identified the factor as galectin-1. Recombinant human galectin-1 (rhGAL-1) showed the same activity at low concentrations (50 pg/ml) that are two orders of magnitude lower than those of lectin activity. A similarly low concentration was also effective in in vivo experiments of axonal regeneration with migrating reactive Schwann cells to a grafted silicone tube after transection of adult rat peripheral nerve. Moreover, the application of functional anti-rhGAL-1 antibody strongly inhibited the regeneration in vivo as well as in vitro. The same effect of rhGAL-1 was confirmed in crush/freeze experiments of the adult mouse sciatic nerve. Because galectin-1 is expressed in the regenerating sciatic nerves as well as in both sensory neurons and motor neurons, we suggest that galectin-1 may regulate initial repair after axotomy. This high activity of the factor applied under nonreducing conditions suggests that galectin-1 may work as a cytokine, not as a lectin.

Molecular nature of spontaneous mutations in mouse lactate dehydrogenase-A processed pseudogenes.

The presence of at least ten mouse LDH-A pseudogenes was demonstrated in the genomic blot analysis, and four different processed pseudogenes have thus far been isolated and characterized. In this report, the nucleotide sequences to two different mouse lactate dehydrogenase-A processed pseudogenes, M11 and M14, were determined and compared with the protein-coding sequences of the mouse and rat LDH-A functional genes. In the pseudogene M11, the sequence of 64 nucleotides from codon no. 257 to 278 was tandemly duplicated. In the pseudogene M14, the sequence of 22 nucleotides from codon no. 68 to 75 was replaced by an inserted repetitive sequence of 242 nucleotides homologous to a mouse truncated R element. The pattern of nucleotide substitutions accumulated in mouse LDH-A pseudogenes M11 and M14, as well as that of pseudogene M10 identified previously, was analyzed, and the substitution frequencies of the C or G at the CG dinucleotide were found to be high.

FLJ10849, a septin family gene, fuses MLL in a novel leukemia cell line CNLBC1 derived from chronic neutrophilic leukemia in transformation with t(4;11)(q21;q23).

A t(4;11)(q21;q23) has been described in 50-70% of cases of infant acute lymphoblastic leukemia and, less frequently, in cases of pediatric and adult acute lymphoblastic leukemia and acute myeloid leukemia (AML). In t(4;11)(q21;q23) leukemias, the AF4 gene has been cloned as a fusion partner of the MLL gene. A human myeloid leukemia cell line, chronic neutrophilic leukemia (CNL)BC1, was established from a peripheral blood specimen of a patient with CNL in leukemic transformation. As with the original leukemia cells, the established line had a t(4;11)(q21;q23). We showed that the MLL gene on 11q23 was fused to the FLJ10849 gene on 4q21. The protein encoded by FLJ10849 belongs to the septin family, sharing highest homology with human SEPT6, which is one of the fusion partners of MLL in t(X;11)(q13;q23) AML. Our results suggest that FLJ10849 might define a new septin family particularly involved in the pathogenesis of 11q23-associated leukemia. The established cell line, CNLBC1, could provide a useful model for analyzing the pathogenesis of MLL-septin leukemias and chronic neutrophilic leukemia.

Relation of serum total cholesterol, serum triglycerides and fasting plasma glucose to colorectal carcinoma in situ.

BACKGROUND: No one has previously examined the relation of serum total cholesterol, serum triglycerides and fasting plasma glucose to colorectal carcinoma in situ. METHODS: A case-control study was performed with 129 cases of colorectal carcinoma in situ and 258 matched controls among examinees undergoing a health check-up in Tokyo from January 1991 to March 1993. RESULTS: There was a significant, positive association between serum total cholesterol levels and the risk of colorectal carcinoma in situ after adjustment for age, sex, body mass index, smoking status and alcohol consumption. Serum triglyceride levels were significantly and positively associated with colorectal carcinoma in situ risk regardless of adjustment for the above covariates. Although there was no clear relation between colorectal carcinoma in situ and fasting plasma glucose levels, a modest increase of colorectal carcinoma in situ risk was observed in the highest category (> or =116 mg/dl) of fasting plasma glucose levels. CONCLUSIONS: The findings suggest a positive association between serum total cholesterol levels and the risk of colorectal cancer, rather than an inverse relation. The strong association with serum triglyceride levels and the weak association with fasting plasma glucose levels support the hypothesis that hyperinsulinaemia may play an important role in colorectal carcinogenesis.

T-cell large granular lymphocytic leukemia occurring after autologous peripheral blood stem cell transplantation.

A 61-year-old man with angioimmunoblastic lymphoma in first complete remission underwent autologous peripheral blood stem cell transplantation. At 1 month post transplant, asymptomatic large granular lymphocytosis developed. The surface marker profile of the cells was CD3+CD8+CD56-CD57+. The disease course was chronic and indolent. The patient remains in complete remission from angioimmunoblastic lymphoma more than 6 months post transplant with persistent large granular lymphocytosis (lymphocyte count, 5-15 x 10(9)/l). Although post transplantation T-cell lymphoproliferative disorders have mostly occurred in allogeneic transplantation recipients and presented as aggressive lymphomas/leukemias, we suggest that chronic indolent T-cell large granular lymphocytic leukemia can occur after autologous stem cell transplantation.

Geranylgeraniol is a potent inducer of apoptosis in tumor cells.

We screened various isoprenoids to find inducers of apoptosis for human leukemia HL-60 cells, and found that GGO (geranylgeraniol) had the most potent apoptosis-inducing activity, as judged from DNA fragmentation in HL-60 cells. The apoptosis-inducing activity of GGO is concentration- and time-dependent. DNA synthesis by HL-60 cells was selectively inhibited on treatment with GGO. Besides HL-60 cells, apoptosis was induced by GGO in various tumor cell lines, including human myeloid multipotential leukemia K562, lymphoblastic leukemia Molt3, and colon adenocarcinoma COLO320 DM.

Ketamine and midazolam kinetics during continuous hemodiafiltration in patients with multiple organ dysfunction syndrome.

OBJECTIVE: To assess the effect of continuous hemodiafiltration (CHDF) on ketamine and midazolam kinetics in multiple organ dysfunction syndrome (MODS). DESIGN AND SETTING: Consecutive clinical study in a general intensive care unit of a university hospital. PATIENTS: Twelve adult patients with MODS requiring CHDF. MEASUREMENTS AND RESULTS: A total of 68 samples were collected during CHDF for ketamine, norketamine, and midazolam assays. The clearance values for ketamine and norketamine were 10.8 +/- 6.6 and 10.9 +/- 11.5 ml/min and their daily extractions were 21.4 +/- 7.1 and 10.2 +/- 11.5 mg/day, respectively. Midazolam was not eliminated through the filter during CHDF. There were no changes in Ramsay Sedation Score or Glasgow Coma Scale during CHDF. CONCLUSIONS: Small fractions of ketamine and norketamine were eliminated during CHDF in MODS. Midazolam was not eliminated during CHDF. CHDF did not affect the sedation using ketamine and midazolam even in MODS patients.

The initial distribution volume of glucose rather than indocyanine green derived plasma volume is correlated with cardiac output following major surgery.

OBJECTIVES: To determine whether the initial distribution volume of glucose (IDVG) rather than plasma volume or blood volume is correlated better with cardiac output during the 4 days following major surgery. DESIGN AND SETTING: Prospective clinical investigation in the general intensive care unit of a university hospital. PATIENTS AND METHODS: 31 consecutive patients who underwent radical surgery for esophageal carcinoma were enrolled. Continuous thermodilution cardiac output monitor was placed in the operating room. Indocyanine green (ICG; 25 mg) and glucose (5 g) were administered simultaneously to calculate IDVG and plasma volume determined using the ICG dilution method. Blood volume was also calculated from plasma volume ICG and hematocrit. Those volumes were measured on admission to the ICU and daily on the first 3 postoperative days. The relationships between each volume and cardiac index (CI), and between routine clinical variables and CI were evaluated. RESULTS: IDVG had a linear correlation with CI in the early postoperative days (r = 0.71, n = 124, p < 0.000001). Measurements of neither the plasma volume nor the blood volume yielded a better correlation with CI than did IDVG (r = 0.45, n = 124, p < 0.000001, and r = 0.23, n = 124, p < 0.01, respectively). No correlation was found between pulmonary artery wedge pressure and CI or between central venous pressure and CI. CONCLUSIONS: Our results indicate that IDVG rather than intravascular volume is correlated with cardiac output. We suggest that IDVG has potential as an alternative indicator of cardiac preload following major surgery.

IL-1 beta enhances neurite regeneration from transected-nerve terminals of adult rat DRG.

We clarified the roles of IL-1 beta in peripheral neural regeneration after axotomy in a three-dimensional collagen gel culture system ranging from a single neurone to a dorsal root ganglion (DRG) explant with its associated nerve bundles. Application of 30 U/ml IL-1 beta to the culture systems clearly enhanced neural regeneration. This regeneration was evident in transected nerve terminals of DRG explants with or without associated nerve bundles, but not in dissociated single neurones. Neural survival was not affected by IL-1 beta in any of these culture systems. These results suggest that IL-1 beta stimulates surrounding non-neuronal cells to secrete neurotrophic factors, thus enhancing neurite regeneration from transected nerve terminals in cultured adult DRG explants.

Expression of the Gfi-1 gene in HTLV-1-transformed T cells.

Human T-cell leukemia virus type I (HTLV-I) is the causative agent of adult T-cell leukemia/lymphoma (ATLL) and immortalizes human T cells interleukin-2 (IL-2)-dependently in vitro. Protracted culture of HTLV-I-infected T cells enables them to grow IL-2-independently. Although acquisition of IL-2-independent growth has been correlated with activation of signal transducers and activators of transcription (STATs), the precise mechanism of IL-2-independent growth is unknown. We found that expression of the Gfi-1 (growth factor independence-1) gene was elevated in most HTLV-I-transformed IL-2-independent cell lines but in few HTLV-I-infected IL-2-dependent cell lines. We also found elevated expression of Gfi-1 in fresh leukemic cells of ATLL patients. Although expression of Gfi-1 is correlated with activation of STAT3, induction of the dominant negative form of STAT3 in the HUT102 cell line does not alter the level of Gfi-1 expression. Furthermore, MT2 cells treated with Gfi-1 antisense oligonucleotide had reduced [3H]thymidine uptake compared with MT2 cells treated with Gfi-1 sense oligonucleotide. These findings indicate that Gfi-1 activation is involved in the IL-2-independent growth of HTLV-I-transformed T cells in vitro and in the development of ATLL in vivo, but is not induced by STAT activation.

Induction of apoptosis in various tumor cell lines by geranylgeraniol.

We have found that GGO is a potent inducer of apoptosis in various human tumor cell lines, including a myeloid multipotential leukemia K562 cell line which is resistant to the induction of apoptosis by various apoptosis-inducing agents and conditions. Polyprenylalcohols with isoprenyl units shorter than the geranylgeranyl group or longer than farnesyl group were less effective in inducing apoptosis in myeloid leukemia HL-60 cells and polyprenylketones had no apoptosis-inducing activity. The fragmentation of DNA in HL-60 cells by GGO was so rapid, no significant effect on the cell cycle was observed. [3H]GGO was taken up by HL-60 cells in 3 h and was incorporated into several proteins. The expression of c-myc and bcl-2 decreased significantly within 3 h of the start of the treatment of HL-60 cells with 50 microM GGO.

Comparative therapeutic evaluation of intrathecal versus epidural methylprednisolone for long-term analgesia in patients with intractable postherpetic neuralgia.

UNLABELLED: BACKGROUND AND OBJECTIVES The goal of this study was to evaluate the analgesic effects of intrathecal versus epidural methylprednisolone acetate (MPA) in patients with intractable postherpetic neuralgia (PHN). METHODS: We studied 25 patients with a duration of PHN of more than 1 year. The patients were randomly allocated to one of two groups: an intrathecal group (n = 13) and an epidural group (n = 12). Sixty milligrams of MPA was administered either into the intrathecal or the epidural space four times at 1-week intervals depending on the treatment group. Continuous and lancinating pain and allodynia were evaluated by a physician unaware of group assignment with a 10-cm visual analogue scale before treatment, at the end of treatment, and 1 and 24 weeks after treatment. In addition, cerebrospinal fluid (CSF) was obtained for measurement of interleukin (IL)-1beta, -6, and -8 and tumor necrosis factor-alpha before and 1 week after treatment. RESULTS: We found marked alleviation of continuous and lancinating pain and allodynia in the intrathecal group (P < .001). The improvements were much greater in the intrathecal group than in the epidural group at all time points after the end of treatment (P < .005). IL-8 in the CSF decreased significantly in the intrathecal group as compared to the epidural group at the l-week time point (P < .01), whereas the other cytokines were undetectable. CONCLUSIONS: Our results suggest the effectiveness of intrathecal as compared to epidural MPA for relieving the pain and allodynia associated with PHN. Also, our findings, together with the decrease in IL-8, may indicate that intrathecal MPA improves analgesia by decreasing an ongoing inflammatory reaction in the CSF.

The modified method of two-step differential extraction of sperm and vaginal epithelial cell DNA from vaginal fluid mixed with semen.

This investigation was undertaken as an efficient method for isolating sperm DNA from a mixed fluid sample which contains vaginal epithelial cells in a greater amount. The modified method of the two-step differential extraction procedure was found to be suitable for separating sperm DNA and vaginal epithelial cell DNA from the mixed stains. As the first step of digestion, vaginal epithelial cells in the mixed stains were lysed with Proteinase K and SDS, and sperm heads remaining in the lysed solution were collected by centrifugation. As the second step digestion, the sperm heads were lysed with the buffer containing Proteinase K, SDS and DTT as reducing agent. DNA fractions extracted from the two lysed solutions were enriched, one with sperm DNA and the other with vaginal epithelial cell DNA. MCT118(D1S80), ApoB VNTR and HLADQ alpha types of sperm DNA were detected and were confirmed by matching with corresponding male blood DNA. In the case of vaginal secretion mixed with semen of two males, the mixture of MCT118 types of the two males was detected in sperm DNA fraction.

Endobronchial involvement with Waldenström's macroglobulinemia.

Although many cases of Waldenström's macroglobulinemia (WMG) with pulmonary involvement have been reported, endobronchial involvement has not been described. We report an unusual case of WMG wherein endobronchial invasion of tumor cells narrowed the airways and caused coughing and wheezing.

Development of interferon-alpha resistant subline from human chronic myelogenous leukemia cell line KT-1.

OBJECTIVE: Interferon-alpha (IFN-alpha) is one of the most effective therapeutic agents for a number of hematological malignancies, including chronic myelogenous leukemia (CML). Nevertheless, its efficacy is limited because of the development of resistance to IFN-alpha therapy. Previously, we established the novel human CML cell line KT-1, which is sensitive to the antiproliferative effects of IFN-alpha. Here, we report the establishment of an IFN-alpha-resistant subline, KT-1/A3R alpha 1000, by culturing KT-1/A3 cells (IFN-alpha-sensitive subline of KT-1) with increasing concentrations of IFN-alpha, in order to analyze the mechanism of acquisition of IFN-alpha resistance in CML cells after IFN-alpha therapy. SUBJECTS AND METHODS: We developed an IFN-alpha-resistant tumor cell variant, KT-1/A3R alpha 1000, from the KT-1/A3 cell line by culturing cells with increasing concentrations of IFN-alpha. This subline was examined for its ability to proliferate and its resistance to apoptosis in high concentrations of IFN-alpha. The induction of the ISGF3 complex in response to IFN-alpha alpha in KT-1/A3R alpha 1000 was compared with that in the parental cell. RESULTS: The levels of interferon-stimulated gene factor 3 components (STAT1, STAT2, and p48) proteins and STAT2 tyrosine phosphorylation induced after IFN-alpha treatment were unchanged, but formation of the ISGF3 complex was remarkably reduced in KT-1/A3R alpha 1000 cells compared to parental cells. CONCLUSION: The KT-1/A3R alpha 1000 subline is a useful model for studying the mechanism of IFN-alpha resistance after IFN-alpha therapy.

Analgesic effect of a herbal medicine for treatment of primary dysmenorrhea--a double-blind study.

We evaluated the analgesic effect of Toki-shakuyaku-san (TSS) in women who had a combination of "deficiency," of "Yin," "cold," and "stagnated blood" syndromes, and were suffering from dysmenorrhea. A diagnostic scoring system was used for determination of these conditions. We treated patients with either TSS or placebo during 2 menstrual cycles with a double-blind technique, and we followed them for 2 additional cycles. A significant alleviation of dysmenorrhea was observed in patients treated with TSS as compared to those treated with placebo. Our results suggest that TSS is effective for treatment of dysmenorrhea in patients with the above-mentioned conditions.

[Superior sagittal sinus thrombosis following L-asparaginase therapy of acute lymphoblastic leukemia].

A 48-year-old female received serial combination chemotherapy including L-asparaginase (L-ASP) for acute lymphoblastic leukemia. After administration of L-ASP, the prothrombin time and activated partial thromboplastin time were prolonged, while fibrinogen and antithrombin III levels markedly decreased, so she was given fresh frozen plasma (FFP). But subsequently, she developed cerebral infarction in the left parietal region and further hemorrhagic infarction in the right parietal region, and died. Autopsy revealed superior sagittal sinus thrombosis and bilateral cerebral infarction, but no obvious thrombus in other organs. Coagulopathy following L-ASP therapy is well-known. In this case, the coagulation studies at the first attack showed that the plasma protein levels of coagulation and fibrinolysis factors decreased in spite of administration of FFP. Fibrin-fibrinogen degradation products (FDP) slightly increased. However there were no significant abnormalities in the platelet count, nor soluble fibrin monomer, which suggested no evidence of disseminated intravascular coagulation. Thus, these findings suggest that L-ASP might be associated with the pathogenesis of thrombosis in this case.

[Successful treatment of CML in accelerated phase with mithramycin].

The patient, an 18-year-old male, was admitted on May 17, 1988, because of high-grade fever, neuralgia and generalized lymphadenopathy. Bone marrow examination revealed a large number of small nests with myeloid blastic cells negative for both peroxidase and TdT activity. Ph1 chromosome and bcr rearranged fragment were positive. On a diagnosis of CML in the accelerated phase, treatment was started with standard BHAC-DMP and vincristine. However, fever still persisted and hematological improvement could not be obtained. From September 20, 1988, mithramycin was given at 25 micrograms/kg every three days. No fever was noted and the NAP score decreased. However, fever reappeared despite the continuing treatment. Combination use of vincristine (1.0 mg/week) and mithramycin (25 micrograms/kg/week) was then begun, and the fever immediately disappeared. After mithramycin administration, a transient marked increase of neutrophils appeared in the peripheral blood, suggesting the induction of differentiation. After then, a complete remission was obtained. A transient disappearance of Ph1 chromosome by the chemotherapy was noticed. He has remained in the chronic phase of CML for one year. In conclusion, combination use of vincristine and mithramycin may be useful in the treatment of the myeloid blast crisis.

[Subcutaneous panniculitic T-cell lymphoma with chromosomal abnormalities and large granular lymphocytes morphology].

A 72-year-old woman was admitted because of a subcutaneous hip tumor. A biopsy specimen of the tumor showed a mixture of medium-sized and large lymphocytes infiltrating the subcutaneous fat tissue with a lobular panniculitis-like pattern--a histologic feature of subcutaneous panniculitic T-cell lymphoma (SPTCL). May-Grünwald-Giemsa-stained cytospin slides of freshly isolated neoplastic cell explants showed that the cells had the characteristics of large granular lymphocytes. Immunophenotypic analysis showed that the cells expressed CD56--a natural killer-associated antigen--as well as the cytotoxic T-cell phenotype CD3+ CD4- CD8+. Southern blot analysis revealed rearrangement bands of the TCR-beta chain gene. Chromosome analysis showed complex abnormalities including t(1;6) (q11; p21). The present case may shed some light on the origin and pathogenesis of SPTCL.