Washout studies of 11C in rabbit thigh muscle implanted by secondary beams of HIMAC.

Heavy ion therapy has two definite advantages: good dose localization and higher biological effect. Range calculation of the heavy ions is an important factor in treatment planning. X-ray CT numbers are used to estimate the heavy ion range by looking up values in a conversion table which relates empirically photon attenuation in tissues to particle stopping power; this is one source of uncertainty in the treatment planning. Use of positron emitting radioactive beams along with a positron emission tomograph or a positron camera gives range information and may be used as a means of checking in heavy ion treatment planning. However, the metabolism of the implanted positron emitters in a living object is unpredictable because the chemical forms of these emitters are unknown and the metabolism is dependent on the organ species and may be influenced by many factors such as blood flow rate and fluid components present. In this paper, the washout rate of 11C activity implanted by injecting energetic 11C beams into thigh muscle of a rear leg of a rabbit is presented. The washout was found to consist of two components, the shorter one was about 4.2 +/- 1.1 min and the longer one ranged from 91 to 124 min. About one third of the implanted beta+ activity can be used for imaging and the rest was washed out of the target area.

[Intra-arterial chemotherapy via reservoir system for advanced bladder cancer and prostate cancer].

A clinical study was performed on the efficacy of intra-arterial chemotherapy using a reservoir system for advanced urological malignancies. The reservoir system was indwelted in the femoral subcutaneous layer by Seldinger's method. Fifteen cases of inoperable complicated advanced bladder cancer and 10 cases of postoperative local recurrent bladder cancer were administered intra-arterial chemotherapy using a reservoir system. Then, 23 cases of local relapsed prostate cancer and two cases of endpocrine-resistant prostate cancer were administered the chemotherapy. The administered anti-cancerous agents were methotraxate, cis-platinum and adriamycin, then 5-FU or carboplatin was administered as maintenance therapy. The mean number of courses of chemotherapy was six for bladder cancer and four for prostate cancer. During stabilization of the local lesion, no distant deterioration was recognized. The overall clinical efficacy was a positive response (PR) and no change (NC): for 18 and 7 cases of bladder cancer, and 11 and 14 cases of prostate cancer, respectively. The median duration of stabilization was 23 months for bladder cancer and 12 months for prostate cancer. The adverse effects were fever than those with systemic chemotherapy.

[Intraarterial chemotherapy via reservoir system for far-advanced bladder and prostate cancer].

A clinical study was performed on the efficacy of intraarterial chemotherapy using reservoir system for far-advanced urological malignancy. The reservoir system was indwelled in the femoral subcutaneous layer using Seldinger's method. Fifteen cases with inoperable complicated advanced bladder cancer and ten cases with postoperative local recurrent bladder cancer received intraarterial chemotherapy using the reservoir system. Then, 23 cases with local relapsed prostate cancer and two cases with endocrine-resistant prostate cancer received chemotherapy. The administered anti-cancerous agents were methotrexate, cis-platinum and adriamycin, and 5-FU or carboplatin were administered as maintenance therapy. The mean courses of chemotherapy were six for bladder cancer and four for prostate cancer. During stabilization of the local lesion, no distant deterioration was recognized. Overall clinical efficacy was as follows: PR:18 cases and NC:7 cases for bladder cancer; then, PR:11 cases and 14 cases for prostate cancer. The median duration of stabilization was as follows: 23 months for bladder cancer and 12 months for prostate cancer. Complications were fewer than with systemic chemotherapy.

[Fleroxacin treatment for acute uncomplicated cystitis in women: comparison of 3-day and 7-day therapy].

The clinical efficacy of fleroxacin (FLRX), a new fluoroquinolone, for acute uncomplicated cystitis (AUC) in women was assessed. Two regimens, 3-day and 7-day courses of FLRX, 200 mg once a day, were compared. Clinical and bacteriological efficacy were evaluated after the therapy, and recurrence rate was evaluated 1 week and 4 weeks after termination of the therapy. Of 136 registered subjects, 35 in the 3-day group and 47 in the 7-day group were evaluated. According to the criteria of Japanese UTI Committee (3rd edition), the rate of excellent results was significantly higher in the 7-day group (78.9%) than in the 3-day group (48.6%), but the overall clinical efficacy rate was similar being 100% and 97.9%, respectively. Although no recurrence was seen 1 week after the therapy in either group, recurrence was seen in 14.3% and 7.4% of the cases in the 3-day and 7-day groups, respectively, 4 weeks after the therapy. Adverse reactions were observed in 2 and 3 cases in the 3-day and 7-day groups, respectively. Both 3-day and 7-day regimens of FLRX treatment showed good efficacy. Although the 7-day treatment was superior to the 3-day treatment as to high rate of excellent results and low rate of recurrence, the 3-day treatment was concluded to be sufficient for AUC.

Bovine lactoferrin stimulates the phagocytic activity of human neutrophils: identification of its active domain.

Bovine LF (bLF) at concentrations in the range of 50-250 micrograms/ml enhanced the phagocytic activity of human neutrophils as determined by measuring the incorporation of FITC-labeled latex beads by flow cytometry. The stimulatory effect of bLF was not abrogated by hydrolysis with pepsin. Bovine lactoferricin (bLFcin), which is a bactericidal fragment purified from a pepsin hydrolysate of bLF (bLFH), also enhanced the phagocytic activity, whereas, in contrast, the fraction of bLFH depleted of bLFcin showed no stimulatory effect. The phagocytosis-enhancing activity of bLF still remained after washing the neutrophils, following exposure to bLF. Also, bLF pretreatment of the latex beads stimulated their uptake. These results demonstrate that bLF is effective in promoting the phagocytic activity of human neutrophils. This activity appears to be due to its bLFcin domain and may involve dual mechanisms of direct binding to neutrophils and opsonin-like activity.

Arbutin increases the pigmentation of cultured human melanocytes through mechanisms other than the induction of tyrosinase activity.

We assessed the effects of arbutin on the pigmentation of cultured normal human melanocytes. As indicated by a cell-blotting assay, arbutin at concentrations in the range of 0.5-8 mM increased the pigmentation of the cultured melanocytes, while kojic acid at concentrations in the range of 0.5-4 mM decreased the pigmentation. The pigmentation-augmenting effect of arbutin was further confirmed by the results of a cell-pelleting assay, the traditional method of assessment. Treatment of the cells with arbutin increased the melanin content of the cells and the protein content as well. On the other hand, the tyrosinase activity in the cells was reduced by arbutin treatment. The levels of transcription of tyrosinase and tyrosinase related protein-1 genes were not affected by arbutin treatment as indicated by a semi-quantitative reverse transcription-polymerase chain reaction assay. These results demonstrate that arbutin promotes an increase in pigmentation of cultured human melanocytes that is not mediated by augmented tyrosinase activity.

Immunomodulatory effect of bovine lactoferrin pepsin hydrolysate on murine splenocytes and Peyer's patch cells.

The effects were examined of a pepsin hydrolysate of bovine lactoferrin on the proliferation of murine splenocytes. The hydrolysate enhanced [3H]thymidine uptake by splenocytes, but undigested bovine lactoferrin exerted an inhibitory effect. The hydrolysate had the ability to inhibit the blastogenesis that was induced by mitogens such as concanavalin A, phytohemagglutinin, and lipopolysaccharide; inhibition was similar to that with undigested lactoferrin. These results suggested that the hydrolysate contained both immunostimulatory and immunoinhibitory peptides. The stimulatory effect of the hydrolysate in the absence of mitogens was then explored in more detail using nonadherent splenocytes. The proliferative response of splenocytes to the hydrolysate was much greater in the fraction that was enriched with B cells than in the fraction that was enriched with T cells. The hydrolysate did not affect thymocyte proliferation. These data indicated that the adherent cells resembling macrophages and found among the splenocytes were not the target cells of the hydrolysate. The stimulatory effect of the hydrolysate was due to the activation of B cells by the hydrolysate and enhanced immunoglobulin production by splenocytes. Because the hydrolysate also enhanced the proliferation and immunoglobulin A production of Peyer's Patch cells, the immunostimulatory effect of the hydrolysate in vivo was examined using mice that had been orally immunized with cholera toxin. The concentrations of immunoglobulin A conjugated against cholera toxin in bile and in the intestinal contents of mice fed liquid diets containing 1% (wt/vol) lactoferrin hydrolysate were greater than those of mice fed control diets. This result suggested that the use of the lactoferrin hydrolysate is beneficial to enhance mucosal immunity.

[Diagnosis and follow-up of prostate cancer patients using prostate specific antigen (PSA)].

An international standard of prostate specific antigen (PSA) assays was constructed and prognosis of the patients with prostate cancers showing gray zone PSA was studied. For lower levels of serum PSA (< 50 ng/ml), the conversion formula to that of Tandem-R PSA from other assays was presented. Furthermore, based on the standards of Stanford Reference and Markit-MPA, conversion rates to this international standard from the conventional PSA assays were also obtained. Patients' cancer-specific survival was found to be significantly better in the gray zone group. Further studies to obtain higher specificity such as using free or complex rate in total PSA is necessary.

Lactoferrin as a suppressor of cell migration of gastrointestinal cell lines.

The effects of lactoferrin (Lf), an iron-binding glycoprotein, on cell migration were investigated. Lf inhibited the cell migration of three gastrointestinal cell lines (Caco-2 cells, AGS cells, and IEC-18 cells) in vitro. Both iron-saturated (holo) and iron-depleted (apo) Lf showed this inhibitory effect. Chelation of iron in the culture medium by desferrioxamine did not affect the activity of either form of Lf. A pepsin hydrolysate of Lf exhibited effectiveness similar to that of intact Lf. These results demonstrate a novel activity of Lf and suggest a potential role for this molecule in gastrointestinal wound healing, which is independent of its iron-binding capacity.

Beta-lactoglobulin suppresses melanogenesis in cultured human melanocytes.

The effects of whey proteins from bovine milk on melanogenesis in cultured human melanocytes were examined. Among the major protein components of milk whey including beta-lactoglobulin (BLG), alpha-lactalbumin, serum albumin, and IgG, only BLG exhibited the depigmenting effect at a concentration of 1 mg/ml. Also, BLG suppressed the activity of tyrosinase in these cells. Retinol, to which BLG is known to bind, slightly increased the pigmentation of the cells at concentrations in the range of 1-100 nM, and retinoic acid, a metabolite of retinol, exhibited a strong pigmentation-promoting effect within the same concentration range. Treatment of the cells with 1 mg/ml BLG completely abrogated the pigmentation induced by these A vitamins. These results demonstrate a novel biological activity of BLG and suggest that this activity is dependent on its ability to bind retinol.

Kappa-casein suppresses melanogenesis in cultured pigment cells.

The effects of bovine milk proteins on melanogenesis in B16 cells were examined. Both whey protein isolate and casein exhibited depigmenting properties. Among the major protein components of milk--including beta-lactoglobulin, alpha-lactalbumin, alpha-, beta-, and kappa-casein--only kappa-casein exhibited the depigmenting effect. However, the carboxyl terminal peptide of kappa-casein, glycomacropeptide, did not show this activity. Also, kappa-casein promoted the proliferation of the cells and inhibited the activity of tyrosinase in the cells. These results indicate that kappa-casein acts as a melanogenesis-suppressing modulator.

Effects of lactoferrin and lactoferricin on the release of interleukin 8 from human polymorphonuclear leukocytes.

Human or bovine lactoferrin (LF) and lactoferricin (LFcin), a peptide derived from the N-terminal region of LF, each have the ability to stimulate the release of neutrophil-activating polypeptide interleukin 8 (IL-8) from human polymorphonuclear leukocytes (neutrophils, PMNs). This finding suggests that LF and LFcin may both function as immunomediators for activating the host defense system. A basic peptide, protamine, exerted the same effect as that of LF and LFcin, suggesting the importance of the basic nature of LF and LFcin in acting as an inducer of IL-8 release from PMNs.

[A case of soft renal calculi with xanthogranulomatous change].

A case of soft renal calculi with xanthogranulomatous change is reported. A 37-year-old female visited our hospital on February 4, 1992 complaining of frequency of urination and right lower abdominal pain. Under the clinical diagnosis of right renal calculi, extracorporeal shock wave lithotripsy was attempted, but no sign of destruction was observed. Right pyelolithotomy was performed on June 8, 1992. Several soft calculi were removed from the right renal pelvis. Microscopic examination showed that the calculi were surrounded by a cell layer including foam cells and giant cells. In addition, the calculi revealed positive reaction by several stainings (alcian-blue, PAS and muticarmin), which showed that the calculi were mainly composed of mucopolysaccharides. We discussed this disease in terms of symptoms, diagnosis, treatment and mechanism in comparison with previous reports.

Effects of lactoferrin and its peptides on proliferation of rat intestinal epithelial cell line, IEC-18, in the presence of epidermal growth factor.

The cell growth-stimulating activity of lactoferrin (LF) in combination with epidermal growth factor (EGF) was evaluated by using a rat intestinal epithelial cell line, IEC-18. LF was found to be more effective than EGF for inducing an increase in cell numbers when cultured for over 6 days using a medium containing 0.2% fetal calf serum (FCS), although the 3H-thymidine incorporation-stimulating activity of EGF was more potent than that of LF. A synergistic effect of LF and EGF was observed in both cell proliferation and DNA synthesis assays. The increase in cell numbers when stimulated with LF plus EGF corresponded to about 5 times that of the control. Iron was not required for manifestation of these effects of LF. On the other hand, iron-saturated transferrin (TF) had cell-growth-stimulating activity, but iron-free TF did not, either in the presence or absence of EGF. These results indicate that LF induces cell proliferation by a mechanism distinct from that of TF. A pepsin-generated hydrolysate of LF (LFH) had an activity similar to that of undigested LF, and a peptide with cell-growth-stimulating activity from bovine LFH was isolated by monitoring its effects in combination with EGF on DNA synthesis in IEC-18 cells. Sequence analysis indicated that the peptide has the structure Ala-Glu-Ile-Tyr-Gly-Thr-Lys-Glu-Ser-Pro-Gln-Thr-His-Tyr-Tyr, corresponding to residues 79-93 of bovine LF.

[Clinical evaluation of TANDEM PSA in Japanese cases and comparison with other methods].

Clinical evaluation of TANDEM PSA which is the most frequently used prostate specific antigen (PSA) assay method in the world and a comparison with other methods were performed in Japanese cases in a cooperative research fashion. The minimum detectable level of the method was found to be 0.50 ng of PSA in one ml of serum and 1.9 ng/ml was regarded as the upper normal value in Japanese males. The distribution of serum PSA showed a significant difference between the benign prostate hypertrophy (BPH) cases and patients with stage C or D prostate cancer. The sero-diagnosis prostate cancer at an early stage with the TANDEM PSA was difficult. The correlation to other methods of PSA detection was very high. Furthermore, the clinical use of the method in following-up the clinical course of prostate cancer patients was very useful. These findings suggested that the PSA detection using TANDEM PSA is applicable even in Japanese cases although the upper cut-off level is decreased.

[Clinical study of neoadjuvant intraarterial chemotherapy (I A-MAC) for advanced bladder cancer].

The clinical efficacy of intraarterial neoadjuvant chemotherapy with MTX, ADR and CDDP in 88 patients with advanced bladder cancer (> or = T2 or TCC G3) was evaluated. The intraarterial chemotherapy was performed using Seldinger's method. The three drugs were administered at the same time among 60 patients. In sixteen patients MTX was administered intravenously before intraarterial infusion of ADR and CDDP. In twelve patients, a high dose of CDDP (more than 100 mg/m2) was administered. The mean regression rate was 70%. That of the high-dose CDDP group was rather superior to the other two groups. However, the adverse reaction and duration of recovery were inferior to those of the other two groups. When intraarterial chemotherapy is performed as a neoadjuvant chemotherapy, care must be taken to determine the suitable dose.

Changes in nerve growth factor content of the submaxillary gland in the genetically dystrophic (mdx) mouse.

We evaluated the nerve growth factor (NGF) contents in the submaxillary gland of the mdx mouse, a model for Duchenne muscular dystrophy (DMD), and found that the NGF and NGF mRNA contents in this organ, where extraordinarily high amounts of NGF are synthesized and stored independently of development or maintenance of the nervous system, were markedly elevated in the male mdx mouse at 8 and 11 weeks of age. However, the NGF content of this organ in 4-week-old male mdx mice was lower than that of control mice although statistical significance of difference was not observed. In the mdx female mouse, the submaxillary NGF content was significantly lower than that of the normal mouse at 4 weeks of age, but was similar to that of the normal at 8 and 11 weeks of age. The amounts of epidermal growth factor (EGF), another protein that is known to be sexually and developmentally regulated in the mouse submaxillary gland like NGF, was found to be also significantly increased in this organ of the male mdx mouse at 8 and 11 weeks of age, and to be significantly decreased in that of the female mdx mouse at 4 weeks of age. The parallel changes in NGF and EGF contents during development of the submaxillary gland suggest that the mdx mouse suffers from some abnormality in the development of this organ.(ABSTRACT TRUNCATED AT 250 WORDS)

Lactoferrin promotes nerve growth factor synthesis/secretion in mouse fibroblast L-M cells.

Fibroblast cells are known to have an ability to synthesize and secrete nerve growth factor (NGF). To investigate the mechanism of action of the iron-binding protein, lactoferrin (Lf), on cultured animal cells, the effect of bovine Lf (bLf) on NGF synthesis/secretion in mouse fibroblast cells was examined. Both apo- and holo-bLf induced an increase in NGF content in the cell-conditioned medium(CM) of mouse L-M cells, a line derived from L929 fibroblast cells, with similar effectiveness. The increase in NGF content in the CM of L-M cells cultured with bLf was not dependent on the induction of increase in cell numbers, but was due to induction of de novo synthesis of NGF in individual cells by bLf. Human Lf(hLf) also increased NGF content. However, apo- and holo-bovine transferrin (bTf) failed to stimulate the NGF synthesis. The time-dependent induction of NGF in L-M cells by bLf was different from that induced by basic fibroblast growth factor (bFGF) and bLf showed an additive effect with bFGF. These results suggest that the induction of NGF synthesis depends on a mechanism different from iron transport or bFGF.

Stimulation by bovine lactoferrin of nerve growth factor synthesis/secretion in mouse L-M cells.

Cultured mouse fibroblasts synthesize and secrete nerve growth factor (NGF). To learn the mechanism of action of an iron-binding protein, lactoferrin (Lf), on cultured animal cells, we have examined the effects of bovine Lf (bLf) on NGF synthesis/secretion in mouse L-M cells, a line derived from L929 fibroblast cells. Both apo- and holo-bLf induced an increase in NGF content in the cell-conditioned medium (CM) with similar effectiveness. Neither apo- nor holo- bovine transferrin (bTf) was effective, indicating that the observed induction of NGF production cannot be attributed to stimulation of iron transport. A basic peptide fragment of bLf (residues 17-41) isolated from a pepsin hydrolyzate of bLf was found to be effective for stimulation of NGF synthesis/secretion in L-M cells. This observation demonstrates that the basic nature of Lf is important for its interaction with fibroblast cells.