Effect of a selective neutrophil elastase inhibitor on early recovery from body water imbalance after transthoracic esophagectomy.

The objective of the study was to evaluate the efficacy of sivelestat, a selective neutrophil elastase inhibitor, on body fluid balance after transthoracic esophagectomy. Esophagectomy with elective lymphadenectomy may induce excessive release of neutrophil elastase, which then promotes vascular permeability and an excessive water shift from the intravascular space to the peripheral compartment. Body fluid imbalance after esophagectomy often leads to circular instability, a decrease of urine output, and a delay in the shift to a diuretic state. The study was designed as a case-control study with a historical control group. A retrospective analysis was performed to examine our hypothesis that sivelestat improves abnormal body fluid retention and prevents subsequent pulmonary complications. To reveal the direct influence of sivelestat on the postoperative course, we avoided using steroids or other diuretic agents. Eighty-eight patients who underwent thoracic esophagectomy with extended lymphadenectomy from 2000 to 2008 were divided into two groups: those treated from 2003 to 2008, who all received postoperative administration of sivelestat (n=60); and those treated from 2000 to 2002, who did not receive sivelestat and were used as the control group (n=28). Both groups received fluid management using the same protocol. The time to reach a diuretic state, time until extubation of the tracheal tube, and development of delayed respiratory dysfunction were compared between the groups using univariate and multivariate analysis. The time until a shift to a diuretic state was significantly shorter after treatment with sivelestat (p<0.0001) and with a shorter operation time (p<0.0001). The tracheal tube was extubated significantly earlier in the sivelestat group (p<0.0001) and the incidence of delayed respiratory dysfunction was also significantly lower (p=0.0028) in this group. Multivariate logistic regression analysis showed that a delay in a shift to a diuretic state was a strong independent risk factor for the time to tracheal extubation (odds ratio 2.539, p=0.0056) and occurrence of delayed respiratory dysfunction (odds ratio 1.989, p=0.0104). Sivelestat treatment was not independently associated with reduced pulmonary complications, but the diuretic state was strongly regulated by sivelestat treatment (odds ratio 0.044, p=0.0003). Thus, administration of sivelestat has a beneficial influence on recovery from body water imbalance through a more rapid return to a diuretic state after esophagectomy, which contributes to prevention of subsequent pulmonary complications.

Histochemical evaluation for the biological effect of menatetrenone on metaphyseal trabeculae of ovariectomized rats.

To evaluate the biological effects of vitamin K2 (menatetrenone, MK-4) on ovariectomy (OVX)-induced bone loss, we have examined histological alterations of femoral metaphyses of sham-operated (sham group), ovariectomized (OVX group), and MK-4 dietary-supplemented OVX (MK-4 group; 50 mg/kg per day) female Fischer rats 1, 2, 5, and 8 weeks after OVX. In the first week, rats of the OVX and MK-4 groups showed discontinuous trabeculae compared with sham-operated rats. At 2 weeks after OVX, the OVX rats revealed many large tartrate resistant acid phosphatase (TRAP)-positive osteoclasts, while osteoclasts in the MK-4-treated rats were similar in size to those of the sham group. At 5 weeks, the OVX and MK-4 groups revealed fragmented trabeculae in femoral metaphyses. The cartilage matrix was partially exposed due to stimulated bone resorption in the OVX group, but not in the MK-4 group. After 8 weeks, the OVX rats had little metaphyseal trabeculae, whereas the MK-4-treated rats had maintained short trabeculae. Despite the presence of intense alkaline phosphatase-positive osteoblasts on trabeculae in the MK-4 group, TRAP-positive osteoclasts were flattened without developing ruffled borders. Therefore, MK-4 appeared to lessen the increase in osteoclastic bone resorption induced by OVX, as well as to maintain the accelerated osteoblastic activity. It is of importance to identify the target cells for MK-4 in bone. Autoradiography localized [3H]-labeled MK-4 mainly in osteoblasts and adjacent bone matrices, but not in osteoclasts, indicating that MK-4 targets osteoblasts. Thus, MK-4 appears to target osteoblasts, consequently inhibiting bone loss induced by ovariectomy.

Vitamin K2 (menatetrenone) inhibits bone loss induced by prednisolone partly through enhancement of bone formation in rats.

Vitamin K(2) (K(2), menatetrenone) has been reported to enhance bone formation and inhibit bone resorption in vitro. However, there is no evidence that K(2) enhances bone formation in vivo. The aim of this study was to characterize the effect of K(2) on bone formation in vivo. We carried out two experiments using a prednisolone (pred)-induced bone loss model in male (10-week-old) Fischer rats. Pred was orally administered three times a week. In experiment 1, we compared the degree of bone loss induced by a 4 week treatment (30 or 100 mg/kg) and an 8 week treatment (3, 10, or 30 mg/kg) with pred by peripheral quantitative computed tomography (pQCT). At 4 weeks, total bone mineral density (BMD) was decreased only with the 100 mg/kg pred treatment. At 8 weeks, total BMD was significantly reduced at >10 mg/kg pred. In experiment 2, we investigated the effect of K(2) on bone loss induced by 3 and 30 mg/kg pred. K(2) (15 mg/kg) was given to rats as a dietary supplement for 8 weeks. Intestinal calcium transport (S/M) and total, trabecular, and cortical BMD at the metaphysis and diaphysis were measured, and histomorphometry was performed in diaphysial cross sections. Pred treatment decreased total and trabecular BMD in the proximal metaphysis. A decrease in cortical BMD in the diaphysis was observed in the pred 30 mg/kg group. Pred treatment also reduced mineralizing surface (MS/BS), mineral apposition rate (MAR), and bone formation rate (BFR/BS). The decrease in total and trabecular BMD in the proximal metaphysis, and in cortical BMD in the diaphysis, was inhibited by K(2) treatment. K(2) treatment also inhibited the decrease in MS/BS and BFR/BS induced by 30 mg/kg pred. These results suggest that K(2) prevents bone loss partly through the enhancement of bone formation.

[Effect of combined treatment with vitamin K2 and 1 alpha-(OH)-vitamin D3 on bone loss in ovariectomized rats].

Different types of therapeutic agents for osteoporosis are often simultaneously prescribed for the same patient, but limited experimental findings indicate the significance of combined treatment. In the present study, the inhibitory effect of combined vitamin K2(K2) and 1 alpha-(OH)-vitamin D3(D3) treatment on bone loss was compared to that of K2 or D3 alone in ovariectomized rats. Female rats (20-week-old) were ovariectomized and divided into 4 groups as follows: they were treated for 8 weeks with vehicle, K2 (30 mg/kg), D3 (0.3 microgram/kg) and K2 and D3 (K2 + D3) at the respective doses. K2 was given as a dietary supplement and D3 was orally administered 3 times a week. Bone density of the femurs was measured by peripheral quantitative computed tomography. Ovariectomy resulted in decreased bone density in proximal metaphysis, especially in the trabecular region, and treatment with K2, D3 or K2 + D3 inhibited this decrease. Moreover, in the K2 + D3 group, bone density and mineral content in the trabecular region in proximal metaphysis and cortical bone width in diaphysis were significantly higher than those in the D3 group. Consistent with these observations, bone strength in the femoral midshaft tended to increase only in the K2 + D3 group compared to that in the vehicle group. These findings indicate that combined K2 and D3 treatment is more effective for bone loss than that with K2 or D3 alone.

Imaging of hydroperoxide and hydrogen peroxide-scavenging substances by photon emission.

An imaging system for the hydroperoxide and hydrogen donor was developed by photon emission from the reactive oxygen species-hydrogen donor-KHCO(3)-MeCHO system. Photon intensity (luminance, cd/m(2)) showed a linear correlation with the concentration of some hydroperoxides [hydrogen peroxide (H(2)O(2)), tert-butyl hydroperoxide and methyl ethyl ketone peroxide] and that of hydrogen donors (catechins and anthocyanins). The linear relationship between photon intensity and concentration was observed in polyphenol-rich samples, such as teas, berries and currants. Photon intensity from the H(2)O(2)/polyphenol-rich sample-KHCO(3)-MeCHO system corresponded with H(2)O(2)-scavenging activity, rather than with polyphenol contents. It is possible to classify hydrogen donor species according to absorption features using spectral analysis (gallic acid, E(max) 645 nm, catechins, E(max) 610 nm; anthocyanins, E(max) 690-700 nm). This method is a simple and sensitive detection system for hydroperoxide- and H(2)O(2)-scavenging substances.

Hypothalamic appetite-regulating neuropeptide mRNA levels in cachectic nude mice bearing human tumor cells.

We previously reported that the human melanoma cell line, SEKI, induces severe weight loss in nude mice. In the present study, we examined the expression of weight-regulating neuropeptide mRNAs in the hypothalamus of this cancer cachectic model by using a sensitive quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method and in situ hybridization. mRNA levels of neuropeptide Y (NPY) and corticotropin-releasing hormone (CRH) in the whole hypothalamus were elevated significantly in the SEKI mice as compared with control mice. In situ hybridization showed that NPY and CRH mRNA were upregulated in the arcuate nucleus and the paraventricular nucleus, respectively. There were no significant differences in melanin-concentrating hormone (MCH), orexin (OX), and cholecystokinin mRNA levels between the SEKI and control mice. These results suggest that the NPYergic system is functioning in the rodent model of cancer cachexia; however, the role of the CRHergic system in energy homeostasis remains to be elucidated. This is the first report of the hypothalamic neuropeptide response to cachexia-inducing human cells.

Response of hypothalamic NPY mRNAs to a negative energy balance is less sensitive in cachectic mice bearing human tumor cells.

We selected three human cancer cell lines [human melanoma (SEKI), human melanoma (G361), and human neuroepithelioma (NAGAI)] that have an ability to develop cancer cachexia syndrome with and without accompanying anorexia and examined the hypothalamic levels of mRNAs for neuropeptide Y (NPY), melanin-concentrating hormone, and orexin. The body weight of sham-operated mice continued to increase, while mice of all tumor-bearing groups lost weight. Competitive reverse transcription-polymerase chain reaction analysis showed that, regardless of feeding status, NPY mRNA levels were elevated in all tumor-bearing mice compared with sham-operated mice, although to a lesser degree than weight-matched pair-weight mice. Melanin-concentrating hormone and orexin mRNA in the hypothalamus followed the same pattern as NPY, although most of the differences did not reach statistical significance. These results support the notion that the response of NPY mRNA to a negative energy balance is less sensitive in these rodent models of cancer cachexia.

[Interaction of warfarin and vitamin K2 on arterial thrombotic tendency using a rat aorta loop model].

Vitamin K2(K2), a therapeutic agent osteoporosis, is prohibited for patients with thrombosis who are receiving warfarin (WF). However, because some aged patients with thrombosis have osteoporosis, some patients treated with WF may be administered K2 concomitantly. We investigated here the interaction between K2 and WF on thrombosis in a rat aorta loop model. Administration of WF at 0.58, 0.82 and 1.16 mg/l in drinking water for 7 days decreased the thrombotic rate and increased the death rate, dose-dependently. Therefore in the following study, 0.80 mg/l of WF was used. After 2 days of WF-treatment, 1.5, 14 and 145 mg/kg of K2 was administered for 5 days. The blood coagulation time was markedly prolonged by WF treatment for 7 days and this effect was completely inhibited by all doses of K2. WF treatment significantly decreased the cumulative thrombotic rate for 5 days. Administration of 1.5 and 14 mg/kg of K2 did not influence the WF effect on thrombosis. The thrombotic rate in the 145 mg/kg K2 group was lower than that in the WF-control group, but similar to that in the WF-untreated group. These findings suggest that high dose of K2 reduces the effect of WF on thrombosis but does not enhance the occurrence of thrombosis more than that without WF treatment.

Cisplatin-5-fluorouracil therapy with remarkable effect and 5-year survival for paraaortic lymph node metastases of rectal carcinoma in females: a case report.

A 68-year-old woman was admitted because of a rectal carcinoma with huge paraaortic lymph node metastases. Low anterior resection with regional lymph node dissection was performed, leaving the paraaortic mass. After the operation, cisplatin-5-fluorouracil therapy was used as supplemental chemotherapy. The metastatic lymph nodes shrank remarkably in response to anticancer drugs. We evaluated the effect of chemotherapy as a partial response. The physical condition of the patient was well controlled for more than 4 years until she was admitted again because of cardiac failure accompanied by relapse of abdominal lymph node swelling. She died of cardiac failure 5 years and 3 days after the operation.

Effects of menatetrenone on the decrease in calcium balance induced by vitamin K-deficient diet and sodium loading in rats.

The effects of menatetrenone (2-methyl-3-tetraprenyl-1,4-naphthoquinone, MK-4) on calcium balance were studied in male Sprague-Dawley rats. Experiment 1: Rats in metabolic cages that were fed a vitamin K-deficient diet and injected daily with latamoxef (100 mg/kg, i.p.) were either treated or untreated with MK-4 for 7 days. Daily food intake, urine volume and feces weight were determined, and calcium concentration in these samples was measured. Calcium balance was calculated as the difference between calcium intake and urinary and fecal calcium excretion. Cumulative calcium balance in the vitamin K-deficient group treated with latamoxef was lower than that in normal rats; this balance was significantly improved by MK-4 (1 and 10 mg/kg, s.c.) administered for 7 days. Experiment 2: Rats were fed a vitamin K-deficient diet containing 4.6% sodium chloride for 6 weeks. MK-4 was administered as a dietary supplement. Forty-eight-hour calcium balance, determined once a week, was significantly reduced compared with that of normal rats after 3 and 5 weeks; the balance was restored dose-dependently by MK-4 administration (1 and 10 mg/kg). Experiment 3: Rats were subjected to the same experimental conditions as experiment 2 for 6 weeks, and intestinal calcium transport was determined using an everted gut-sac technique. Calcium transport was reduced by the high sodium, vitamin K-deficient diet, and this reduction was restored by MK-4 administration (10 mg/kg). These results suggest that MK-4 improves the reduced calcium balance by increasing intestinal calcium absorption in these rats.

Effects of menatetrenone on prednisolone-induced bone loss in rats.

This study was carried out to evaluate the effect of menatetrenone, a vitamin K2 with 4 isoprene units, on prednisolone-induced bone loss. Three experiments were performed in rats which received menatetrenone as a dietary supplement. In experiment 1, a soluble form of prednisolone, dissolved in drinking water, was administered to rats at 7 mg/kg/day for 9 weeks. The length, dry weight, and bone density of femurs and tibiae, as well as urinary gamma-carboxyglutamic acid (Gla) content, were significantly lower in the prednisolone-control group than in the intact group. Menatetrenone (17 mg/kg/day) significantly inhibited the decrease in these bone parameters, especially in tibiae, and completely inhibited the decrease in urinary Gla content. In experiments 2 and 3, prednisolone (10 mg/kg), dissolved in cottonseed oil, was given to rats intramuscularly three times a week for 4 and 10 weeks, respectively. In experiment 2, bone length, bone strength and calcium content in the femur were reduced by 4-week prednisolone treatment. These reductions were significantly improved by menatetrenone (21 mg/kg/day). In experiment 3, 10-week prednisolone treatment reduced bone length and the calcium and hydroxyproline content of the femur. Menatetrenone (0.4, 10, and 50 mg/kg/day) significantly inhibited the reduction of calcium content in the femur. These results suggest that menatetrenone may inhibit the bone loss induced by corticosteroid treatment.

Effects of menatetrenone on bone loss induced by ovariectomy in rats.

The effects of menatetrenone, a vitamin K2 homologue, on bone loss induced by ovariectomy in rats were studied in 3 experiments. Menatetrenone was given as a dietary supplement. In experiment 1, at 2 weeks postovariectomy, menatetrenone (10 mg/kg/day given for 2 weeks) inhibited the decrease in bone density of the femoral metaphysis induced by the ovariectomy. In experiment 2, menatetrenone (3 or 30 mg/kg/day given for 6 months) inhibited the decrease in bone strength of the femur and the decrease in calcium and hydroxyproline content of the femoral diaphysis at 6 months postovariectomy. In experiment 3, menatetrenone treatment, at 30 or 100 mg/kg/day for 6 months, protected against the decrease in bone strength and calcium and hydroxyproline content in the bone loss model induced by ovariectomy and calcium-deficient diet. These findings suggest that menatetrenone protects against the bone loss induced by ovariectomy.

[A case of adult onset hypophosphatemic vitamin D resistant osteomalacia].

A case of adult onset hypophosphatemic vitamin D resistant osteomalacia is described. A 40-year-old female who complained of thorax and lumbar pain and gait disturbance was admitted to our hospital on 7 November, 1988. The patient had hypophosphatemia with normal plasma calcium, parathyroid hormone and 25-hydroxy vitamin D3 concentrations, but had decreased tubular reabsorption of phosphate and decreased plasma 1, 25-dihydroxy vitamin D3 concentrations. The iliac crest bone biopsy showed osteomalacic changes. The 99mTc-MDP bone scintigram showed evidence of increased bone turnover with raised plasma alkaline phosphatase concentrations. After treatment with oral 1 alpha-hydroxy vitamin D3 (3-6 micrograms/day) and intravenous or oral phosphate supplement (0.47-1.74g/day), the subjective and clinical findings improved.

[A case of Nasu-Hakola's disease with T2-weighted MRI finding of reduced signal intensity in the thalamus and putamen].

A 30-year-old female received a head injury at the age of 22 years. Subsequently neurological and psychiatric symptoms, such as personality change, urinary incontinence, dementia and gait disturbance developed. On admission, her cognitive function was severely impaired. Brain CT disclosed cerebral atrophy, dilatation of the lateral ventricle and calcification of the basal ganglia. Pathologically membranous structures were recognized in bone marrow. On the basis of these clinical findings, a diagnosis of Nasu-Hakola's disease was made. In this case, a T2-weighted MRI finding of reduced signal intensity in the thalamus and putamen was characteristic. This finding may be related to intracranial calcification.