The potential antioxidant activity of incorporating bacaba (Oenocarpus bacaba Mart.) extract into a nanoemulsion system with baru oil.

The bacaba (Oenocarpus bacaba Mart.) peel corresponds to 15% of the whole fruit and is rich in antioxidants with potential application in product development. In nanotechnology, emulsified formulations such as nanoemulsions stand out for providing modified release and improving the bioavailability of conveyed substances. The aim of this work was to develop nanoemulsified systems from baru oil containing hydroalcoholic extract from the bacaba peel, evaluate their stability and antioxidant potential. After the HLB (Hydrophilic-lipophilic balance) determination of the baru oil, thirty-two formulations were developed, varying the proportions of surfactants, aqueous phase, and baru oil. Of those 32, 16 formed emulsified systems, and the ones with a higher amount of oil (20%) were incorporated with the BPE. The systems were submitted to stability studies to verify their viability. After that, several tests were performed, such as rheological characteristics, hydrodynamic diameter of the droplets, polydispersion index, zeta potential, and antioxidant potential by DPPH and ABTS+ radical scavenging methods. After the studies, two samples remained stable and presented a non-Newtonian pseudoplastic profile with thixotropy, hydrodynamic diameter of less than 200 nm, monodispersity, and negative zeta potential. The BPE showed antioxidant potential, with superior activity when incorporated into the nanoemulsified system. A strong negative correlation was found between the two antioxidant methods, where both demonstrated the same profile of potential antioxidant activity for the extract and formulations. The studied formulation showed that the use of BPE is a viable alternative for the development of new products based on sustainable technologies.

Induction of EMT-like phenotypes by an active metabolite of leflunomide and its contribution to pulmonary fibrosis.

Drug-induced interstitial lung disease (ILD), particularly pulmonary fibrosis, is a serious clinical concern and myofibroblasts have been suggested to have a major role, with it recently being revealed that some of these myofibroblasts are derived from lung epithelial cells through epithelial-mesenchymal transition (EMT). In this study, we examined the EMT-inducing abilities of drugs known to induce ILD clinically. EMT-like phenotypes were induced by A771726, an active metabolite of leflunomide having an inhibitory effect on dihydroorotate dehydrogenase (DHODH). Smad-interacting protein 1 (a transcription factor regulating EMT) and the Notch-signaling pathway but not transforming growth factor-ß was shown to be involved in A771726-induced EMT-like phenotypes. When the cultures were supplemented with exogenous uridine, the A771726-induced EMT-like phenotypes and activation of the Notch-signaling pathway disappeared. Similarly, an A771726 analog without inhibitory activity on DHODH produced no induction, suggesting that this process is mediated through the inhibition of DHODH. In vivo, administration of leflunomide stimulated bleomycin-induced EMT-like phenomenon in pulmonary tissue, and exacerbated bleomycin-induced pulmonary fibrosis, both of which were suppressed by coadministration of uridine. Taken together, these findings suggest that leflunomide-dependent exacerbation of bleomycin-induced pulmonary fibrosis is mediated by stimulation of EMT of lung epithelial cells, providing the first evidence that drug-induced pulmonary fibrosis involves EMT of these cells.

Effects of garlic preparations on the gastrointestinal mucosa.

The effects of garlic preparations, including dehydrated raw garlic powder (RGP), dehydrated boiled garlic powder (BGP) and aged garlic extract (AGE), on the gastric mucosa were determined using a newly established endoscopic air-powder delivery system, which can deliver solid materials directly into the stomach. Among the three preparations, RGP caused severe damage, including erosion. BGP also caused reddening of the mucosa, whereas AGE did not cause any undesirable effects. The safety of enteric-coated garlic products was also determined. Direct administration of pulverized enteric-coated products on the gastric mucosa caused reddening of the mucosa. When an enteric-coated tablet was administered orally, it caused loss of epithelial cells at the top of crypts in the ileum. These results suggest that caution be used with regard to safety and effectiveness when choosing a garlic preparation because some preparations may have undesirable effects, including gastrointestinal problems.

Mean corpuscular volume may be a useful index of risk for colorectal adenoma in middle-aged Japanese men.

OBJECTIVE: It has been reported that alcohol intake and folate deficiency are associated with an increased risk of colorectal adenomas and carcinomas. Mean corpuscular volume (MCV) of red blood cells has been reported to be increased in these conditions. The purpose of this study was to assess the association between MCV and risk of colorectal adenoma. METHODS: The subjects were 497 middle-aged (45-65 yr old) men who underwent both barium enema examination and total colonoscopy. The subjects answered a questionnaire regarding their alcohol consumption history, and their blood samples were analyzed. The subjects were divided into four groups three times: with or without alcoholism, and with or without adenoma according to alcohol intake, and according to the MCV value. Various variables were compared among the groups, and the odds ratios of adenoma were calculated. RESULTS: The MCV was higher in the alcoholic group than in the nonalcoholic group (p < 0.01) and in patients with adenoma than in those without adenoma (p < 0.0001). When the subjects were stratified by alcohol intake, the MCV value had a higher significant difference than alcohol intake, between patients with adenoma and those without adenoma. As for the MCV value, the odds ratio (95% confidence interval) of adenoma was 1.00 (referent); (<92), 1.20 (0.71-1.69); (> or =92 but <95), 2.61 (2.07-3.15); (> or =95 but <98); and 3.62 (2.99-4.25); (> or =98). CONCLUSION: A high MCV value may be used as a simple index of the risk of colorectal adenomas, regardless of alcohol consumption.

Endoscopic diagnosis of lower intestinal lesions of leukaemia and malignant lymphoma.

There have been only a few endoscopic studies with respect to lower intestinal lesions of leukaemia and malignant lymphoma, although there have been many autopsy studies of these lesions. The aim of this study was to clarify these lesions using endoscopy. Colonoscopy was performed on 11 of 341 patients with leukaemia and on 32 of 105 patients with malignant lymphoma for frequent diarrhoea, anal bleeding or abnormal findings on barium enema examination, between April 1984 and September 1994. In eight of the 11 patients with leukaemia on whom endoscopy was performed, nine lesions were found; aphthoid ulcers, small ulcers or large tumours due to leukaemic infiltration were found in five, and colorectal adenoma was found in only one patient. Antibiotic-associated haemorrhagic colitis or pseudomembranous colitis was found in one patient each. In 10 of the 32 patients with malignant lymphoma, 11 lesions were found. The following were found in one patient each: large lymphomatous tumours, a large lymphomatous ulcer, multiple small polypoid lesions, multiple lymphomatous polyposis; and colorectal cancer or adenoma in six patients. However, the autopsy findings in patients with both diseases were mostly pseudomembrane formation or ulcers due to fungal and/or bacterial infection. It is concluded that accurate endoscopic diagnosis of lower intestinal lesions in patients with leukaemia or malignant lymphoma is essential for staging and treatment of these diseases and for determining their prognosis. Most lesions in leukaemia are aphthoid and small ulcers are due to leukaemic infiltration or antibiotics; most lesions in malignant lymphoma are elevated lesions such as cancer, adenoma or lymphomatous lesions as determined by endoscopy. This is in contrast to pseudomembrane formation or ulcers due to fungal and/or bacterial infection which are detected at autopsy.

Tone burst-evoked otoacoustic emissions in cats with acoustic overstimulation and anoxia.

Transient evoked otoacoustic emissions (TEOAE) produced by a 2 kHz tone burst could be detected in 30 out of 37 ears (81% detectability) in 21 cats. The amplitude of tone burst-evoked TEOAE was saturated at a stimulus level between 45 and 50 dB SPL and the latency time of peak amplitude was 6.23 ms on average (5.53-7.28 ms). The effects of pure tone overstimulation and short-term anoxia on the tone burst-evoked TEOAE in cats were evaluated. A permanent detection threshold shift of the TEOAE was confirmed at 24 h and 1 week after the overstimulation at 125 dB SPL. In these cases, damaged first row outer hair cells and inner hair cells were observed over an average length of 3.3 mm (16% of the entire cochlear length) by scanning electron microscopy. These findings suggested that the TEOAE can detect localized cochlear hair cell damage. A temporary detection threshold shift of the TEOAE was observed after the overstimulation at 105 dB SPL, and the threshold shift recovered in 107.5 min on average. In the short-term anoxia trial, the TEOAE amplitude started to decrease 45-90 s after the anoxia and recovered completely when the duration of anoxia was under 1 min. However, the TEOAE amplitude did not recover pre-anoxia values (it remained below 80% of its initial value) after 5 min when the anoxia was over 2 min. These findings demonstrated that the detection threshold and amplitude of the TEOAE were also affected by metabolic changes of the cochlear hair cells. Tone burst-evoked TEOAE are useful for the evaluation of localized histological and functional damage of the cochlear hair cells.

Electron-microscopic and immunohistochemical studies of Langerhans cells and Thy-1-positive cells in mouse tongue epithelium subjected to local hyperthermia.

Local hyperthermia via skin has been used to treat cancer but may suppress local immune responses as a side-effect. To examine effects of heat on immunologically responsive cells in oral mucosa, mouse tongues were heated by an implant system at 43 degrees C for 20 min. The densities of Langerhans cells and Thy-1-positive cells rapidly increased within 3 h after the treatment, then returned to a normal level after 7 days. Electron microscopy confirmed that Langerhans cells in the tongue epithelium formed clusters with lymphocytic cells, suggesting an active response to the hyperthermia.

Plant annexin form homodimer during Ca(2+)-dependent liposome aggregation.

The annexin (p35) was isolated from the fruits of green pepper (Capsicum annum). The partial amino acid sequence of p35 was analyzed. p35 had an endonexin fold as annexin consensus sequences. Purified p35 had other annexin like characters such as strongly bind to phosphatidylserine and phosphatidylinositol, phospholipase A2 inhibition and liposome aggregation. The zero-length crosslinking assay revealed that p35 formed a homodimer during Ca(2+)-dependent liposome aggregation.

Anti-osteopenic effect of nasal salmon calcitonin in type 1 osteoporotic rats: comparison with subcutaneous dosing.

The anti-osteopenic effect of nasal salmon calcitonin (SCT) was investigated in a type 1 osteoporotic model, Wistar rats which were ovariectomized (OVX) at age of 12 weeks, and compared with that of subcutaneous SCT. It was proved that nasal (5, 10, 20 and 40 U/rat) and subcutaneous (5, 10 and 20 U/kg) administration of SCT on alternate days for 3 weeks, starting a week after OVX, prevented the osteopenic changes of tibia and lumbar vertebra; this was proved by physicochemical parameters and histomorphometrically. A clear dose-dependent effect was seen in the trabecular bone volume of a selected regions of the 5th lumbar vertebra, and the ED50s of nasal and subcutaneous SCT calculated were 7.4 U/rat and 3.5 U/kg, respectively. The results indicate that nasal SCT is absorbed efficiently in rats with increased bone turnover to prevent rapidly developing osteopenia and that the administration route is a suitable standard method for chronically giving biodegradable anti-osteoporotic peptides to rats.

Effects of 1 alpha-hydroxyvitamin D3 on lumbar bone mineral density and vertebral fractures in patients with postmenopausal osteoporosis.

The effects of 1 alpha-hydroxyvitamin D3 [1 alpha(OH)D3] on bone mineral density, fracture incidence, and bone metabolism were evaluated by a double-blind, placebo-controlled study. Eighty postmenopausal osteoporotic Japanese women (71.9 +/- 7.3 years, mean +/- SD) were randomly assigned to 1 microgram of 1 alpha(OH)D3 daily or inactive placebo for 1 year. All patients were given supplemental calcium (300 mg of elemental calcium daily). Lumbar (L2-L4) bone mineral density (BMD) determined by dual energy X-ray absorptiometry increased 0.65% with 1 alpha(OH)D3 treatment and decreased 1.14% with placebo (P = 0.037). BMD in both the femoral neck and Ward's triangle did not yield any significant differences between the two groups, whereas trochanter BMD in the 1 alpha(OH)D3-treated group increased 4.20% and decreased 2.37% with placebo (P = 0.055). X-ray analysis demonstrated that new vertebral fractures occurred in two patients with 1 alpha(OH)D3 and in seven patients with placebo. The vertebral fracture rate in the treated group was significantly less (75/1000 patient years) than in the control group (277/1000 patient years; P = 0.029). Hypercalcemia (12.1 mg/100 ml) occurred in one patient receiving 1 alpha(OH)D3; however, the serum calcium level in this patient promptly decreased to the reference range after cessation of the treatment. There were no significant changes in serum creatinine level in either group. A significant increase in urinary excretion of calcium was found but there was no significant change in urinary excretion of hydroxyproline in the treated group. The serum level of bone-derived alkaline phosphatase activity significantly decreased by -26 +/- 26 (mU/ml) after the treatment (P = 0.003).(ABSTRACT TRUNCATED AT 250 WORDS)

Swainsonine augments the cytotoxicity of human lymphokine-activated killer cells against autologous thyroid cancer cells.

OBJECTIVE: Swainsonine (SW), an inhibitor of mammalian Golgi alpha-mannosidase II, blocks the processing of high mannose to complex type oligosaccharides. In this study, the effect of SW on the cytotoxicity of lymphokine-activated killer (LAK) cells against autologous thyroid cancer was investigated. DESIGN: Peripheral blood lymphocytes from patients with thyroid cancer were incubated with recombinant interleukin 2 (100 U/mL) and 0.5 mg/L of SW for 7 days, and thyroid cancer cells obtained from surgical specimens were pretreated with SW (0.5 mg/L) for 18 hours. The cytotoxicity of SW-treated LAK cells against tumor cells tested in a standard 4-hour radioactive chromium Cr 51 release assay. RESULTS: The cytotoxicity of SW-treated LAK cells against autologous thyroid cancer cells was found to be significantly greater than that of standard LAK cells incubated with interleukin 2 alone. The N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester esterase activity of LAK cells, this activity being a cytotoxic factor that is necessary for the lethal hit stage, was also increased by SW treatment. Further, thyroid cancer cells incubated with SW, as compared with nontreated tumor cells, showed much higher susceptibility to LAK killing. CONCLUSIONS: Our results suggest that SW might have potential immunomodulatory properties in the treatment of thyroid cancer.

A suitable culture medium for ossification of embryonic chick femur in organ culture.

To establish a culture medium which allows ossification in organ culture, 9-day-old embryonic chick femurs were cultured in variously supplemented BGJb-HW2 media. Changes of Ca and Pi concentrations in the BGJb-HW2 medium or the 10% addition of chick embryo extract (CEE) did not induce ossification. Furthermore, combinations of the 10% CEE with a high Ca x Pi product or with 5 mM beta-glycerophosphate (beta-GP) or with 10% horse serum plus a high Ca x Pi product often caused pathological abnormalities in the periosteum. On the other hand, BGJb-HW2 medium supplemented with 5 mM beta-GP induced development of ossification. The Ca content of femurs and the diaphysial hydroxyproline content were markedly increased. Histological observation showed a formation of a thick and active periosteum, numerous osteoblastic cells, a sufficient amount of osteoid tissue and well developed calcified trabeculae without any pathological changes. Thus, the organ culture system using this medium was considered to be an appropriate one for studies on osteogenesis in vitro.

[Local hyperthermia using a device made of temperature-sensitive ferrite. The first report: temperature, output characteristics].

The basic study of intracavitary and interstitial hyperthermia was made with the Soft-Heating Method. In this method, a hybrid device composed of non-magnetic metal ring and ferrite rod is used as a heater. Being exposed to external magnetic field, the device produces heat, and its temperature rises to the Curie Temperature of the ferrite rod with self-regulation. The accuracy of the regulation is +/- 0.2 degrees C near the Curie Temperature which can be set up almost freely. In addition to that, the frequency of magnetic field is not so high (20-50 KHz) that there is no damping nor reflection of the field in a human body. Therefore the system seems to be very useful for hyperthermia of deep seated tumor. In this paper, experimental discussion of the possibility of local hyperthermia by the system is made. From the results, it is concluded that the system will be applied to clinical uses.

[Therapeutic effectiveness of vitamin D3 in patients with myelodysplastic syndromes, leukemias and myeloproliferative disorders].

We tried to treat 13 patients with myelodysplastic syndromes (MDS), leukemias and myeloproliferative disorders, with alfacalcidol for their hematological improvement. Eight of them had MDS, 2 acute leukemia (M3, M4), 1 chronic myelogenous leukemia and 2 primary myelofibrosis. All patients were untreated except for 3 patients (PASA, RAEB, AML-M4) who had been treated with mepitiostane, prednisolone and BH.AC-AMP regimen, respectively, prior to alfacalcidol therapy. All patients received alfacalcidol orally for at least one month. The dosage of alfacalcidol ranged from 0.25 to 10 micrograms/day, and the medicine was administrated intermittently when the dosage exceeded 6 micrograms/day to prevent hypercalcemia. The therapeutic effectiveness of alfacalcidol was evaluated according to a criteria by Koeffler (Cancer Treat Rep 69: 1399, 1985) with minor modifications. Three patients (PASA, RAEB, CMML) showed partial response, 3 (RAEB, RAEB in T, AML-M4) minor response and rest of the patients did not respond. The hematological improvement of 6 responders was transient (from 1 to 2 months), however, one patient (PASA) is still responding to alfacalcidol therapy (0.25 microgram/day) for over 12 months. The dysplastic features of hemopoietic cells in the bone marrow showed no noticeable change during the hematological improvement in these responders, suggesting the improvement was obtained as a result of alteration in the proliferation or differentiation of neoplastic clone. None of 13 patients developed hypercalcemia. One patient (AML-M4) became excitable on high dose alfacalcidol (10 micrograms/day). In conclusion, alfacalcidol therapy is effective in some patients with MDS or leukemias and appears worthy especially in the clinical state in which chemotherapy is not indicated.

[A case of retroperitoneal chylous cyst developed after cholecystectomy and choledochotomy].

A 43-year-old woman complaining of upper abdominal pain was referred to our clinic. Well movable mass was detected in the right hypochondria region by palpation. Three years before, she underwent cholecystectomy and choledochotomy for cholelithiasis. After that she had been aware of painless mass in the right hypochondria region, but because of no symptom she regarded it as operative scar and had no treatment. Abdominal CT, ultrasonography, upper GI series and barium enema revealed a retroperitoneal cyst compressing the 3rd portion of the duodenum upward and the vena cava backward. Laparotomy showed an unilocular chylous cyst, 5 cm in diameter in the retroperitoneal cavity. Histologically, the cyst was cystic lymphangioma. Etiologically, the previous operation of cholecystocholedocholithiasis was supposed to be a trigger of the development of the chylous cyst.

[Clonogenicity and BCNU response of subpopulations of human glioma cells sorted according to DNA content].

Malignant gliomas, especially glioblastoma multiforme, are composed of cells that contain various amounts of DNA. We characterized cells dissociated from biopsy specimen of human gliomas by sorting vital cell populations on the basis of DNA content, determined the sensitivity of sorted cells to BCNU, and did the follow-up study using subpopulations cultivated for a long time after sorting on both the DNA histogram and the BCNU response. Seven malignant human gliomas carried in monolayer culture which had three peaks on the DNA histogram were used for this study. To obtain each DNA distribution histogram, cells were stained with chromomycin A 3 and were analyzed with a modified FACS III flow cytometer. Vital DNA was stained with Hoechst 33342 by means of a modification of the procedure of Jovin et al, which involved negligible toxicity to cells. Cells were sorted on FACS III at each peak on the DNA histogram. The colony forming efficiency (CFE) of each population was determined after sorting and surviving fraction was determined after exposure to graded doses (2.5-10 micrograms/ml X 2 hr) of BCNU. Six subpopulations derived from two human glioma cells were cultivated for a long time and reevaluated by both the DNA histogram and the sensitivity to BCNU. The CFEs for each cell population of three peaks on DNA histograms of human glioma cells were similar, except that the CFE for the far left peak (peak 1) in each histogram appeared to be slightly lower in four specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effect of eel calcitonin derivative (elcatonin) in osteoporosis].

Calcitonin has been considered to be useful in the treatment of osteoporosis due to its remarkable suppressive action on bone resorption as well as its putative stimulative effect on bone formation. However, its usefullness, especially the effect for decreasing the loss of bone mass in osteoporosis has not been proved substantially. We have studied this point using eel calcitonin derivative, elcatonin , in relatively low doses with and without calcium supplement. Bone mass was evaluated by microdensitometry of the roentgenograms of the second metacarpal bone. One hundred and thirty cases of postmenopausal and senile osteoporotics were divided into the following four groups and studied for six months: A) calcium and elcatonin treated group, B) calcium treated group, C) elcatonin treated group, D) control group. Elcatonin was administered intramuscularly 10 units once a week, and calcium was given 700 mg for every day. The results of metacarpal densitometry revealed the signs of significant increase of the bone mass in the calcium and elcatonin treated (A) group compared with the control (D) group, while the changes in the other groups (B and C) were not significant. It was shown that elcatonin had the significant analgesic effect for the back pain in the patients. Serum iPTH was not increased and the antibody for elcatonin was not detected in elcatonin treated patients. Although the study period is rather short, necessitating further studies, at present, our study suggests that the use of calcitonin with calcium supplements had the place in the treatment of osteoporosis.

Intrarenal distribution of blood flow and renin in chronic congestive heart failure.

Intrarenal distribution of blood flow was measured by the 133xenon washout curve in 33 patients with heart disease. Plasma renin activity and sodium concentration were also measured on the day when the xenon study was performed. The patients were divided into three groups according to cardiac index: Group I whose cardiac index showed higher than 3.50 1/min/M2, BSA, group II whose index ranged from 2.50 to 3.50, and group III who had lower than 2.50. Total renal blood flow was significantly decreased in group II (p less than 0.001), as compared with normal controls. The percents of the total renal blood flow supplied to component I decreased significantly in group I, II (p less than 0.05) and group III (p less than 0.01). The flow rate in component I decreased significantly only in group II (p less than 0.05) and group III (p less than 0.01). There was a significant increase in the percent distribution of component II in group II (p less than 0.05) and in group III (p less than 0.01). The flow rate of component II showed a slight increase in group I and III. The study of autoradiographs done in dogs with heart failure demonstrated that component I corresponded to a cortical area having a relatively faster flow rate, whereas component II corresponded to the cortical area which was perfused more slowly. Accordingly, component III indicated outer medulla. There was no apparent relation between intrarenal distribution of blood flow and plasma renin activity although the latter tended to be elevated in patients treated with diuretics. In view of the data available it was concluded that outer cortical as well as outer medullary blood flow are decreased in chronic congestive heart failure and that there is no apparent correlation between outer cortical flow and plasma renin activity.