Shosaikoto increases calprotectin expression in human oral epithelial cells.

BACKGROUND AND OBJECTIVE: Oral epithelial cells help to prevent against bacterial infection in the oral cavity by producing antimicrobial peptides (AMPs). A broad-spectrum AMP, calprotectin (a complex of S100A8 and S100A9 proteins), is expressed by oral epithelial cells and is up-regulated by interleukin-1alpha (IL-1alpha). Shosaikoto (SST) is a traditional Japanese herbal medicine that has immunomodulatory effects and is reported to enhance the levels of IL-1alpha in epithelial cells. The purpose of this study was to investigate the effect of SST on the expression of calprotectin and other AMPs through the regulation of IL-1alpha in oral epithelial cells. MATERIAL AND METHODS: Human oral epithelial cells (TR146) were cultured with SST (at concentrations ranging from 10 to 250 microg/mL) in the presence or absence of anti-IL-1alpha or IL-1 receptor antagonist. The expression of S100A8- and S100A9-specific mRNAs was examined by northern blotting. Calprotectin expression and IL-1alpha secretion were investigated by immunofluorescent staining or ELISA. The expression of other AMPs and IL-1alpha was analyzed by RT-PCR and by quantitative real-time PCR. RESULTS: Shosaikoto (25 microg/mL) significantly increased the expression of S100A8- and S100A9-specific mRNAs and calprotectin protein. Shosaikoto increased S100A7 expression, but had no effect on the expression of other AMPs. The expression of IL-1alpha-specific mRNA and its protein were slightly increased by SST. A neutralizing antibody against IL-1alpha or IL-1 receptor antagonist inhibited SST up-regulated S100A8/S100A9 mRNA expression. CONCLUSION: These results suggest that SST increases the expression of calprotectin and S100A7 in oral epithelial cells. In response to SST, up-regulation of calprotectin may be partially induced via IL-1alpha.

Regulation of calprotectin expression by interleukin-1alpha and transforming growth factor-beta in human gingival keratinocytes.

BACKGROUND AND OBJECTIVE: Calprotectin, a heterodimer of S100A8 and S100A9 with antimicrobial properties, is expressed in gingival keratinocytes and plays an important role in innate immunity. Because calprotectin expression is localized in the spinous cell layer of the gingival epithelium, we hypothesized that the expression of calprotectin in keratinocytes is related to the differentiation stage. The aim of the present study was to investigate the relationship between calprotectin expression and keratinocyte differentiation using some factors that regulated its differentiation. MATERIAL AND METHODS: Normal human gingival keratinocytes were isolated from gingival tissues obtained at the extraction of wisdom teeth, and were cultured in serum-free keratinocyte medium supplemented with interleukin-1alpha or calcium, which promote keratinocyte differentiation, and transforming frowth factor-beta (TGF-beta) or retinoic acid, which suppress its differentiation. The expression of S100A8/A9 mRNA and the production of calprotectin in normal human gingival keratinocytes were examined by northern blotting and enzyme-linked immunosorbent assay, respectively. The expression of cytokeratin 14, involucrin and filaggrin (marker proteins of keratinocyte differentiation) was investigated by immunohistochemical staining, and the DNA-binding activity of CCAAT/enhancer binding protein alpha (C/EBPalpha), a transcription factor, was examined by electrophoretic mobility shift assay. RESULTS: The expression of S100A8/A9 mRNA and the production of calprotectin were increased by interleukin-1alpha and calcium, but decreased by TGF-beta. RA inhibited the expression of S100A8/A9 and keratinocyte differentiation, which were induced by interleukin-1alpha. C/EBPalpha DNA-binding activity in normal human gingival keratinocytes was enhanced by interleukin-1alpha and calcium, but suppressed by TGF-beta. CONCLUSION: The present study suggests that calprotectin expression is related to keratinocyte differentiation and that C/EBPalpha is a regulator of calprotectin expression in keratinocytes.

Low-temperature Fourier transform infrared spectroscopy of photoactive yellow protein.

The photocycle intermediates of photoactive yellow protein (PYP) were characterized by low-temperature Fourier transform infrared spectroscopy. The difference FTIR spectra of PYP(B), PYP(H), PYP(L), and PYP(M) minus PYP were measured under the irradiation condition determined by UV-visible spectroscopy. Although the chromophore bands of PYP(B) were weak, intense sharp bands complementary to the 1163-cm(-1) band of PYP, which show the chromophore is deprotonated, were observed at 1168-1169 cm(-1) for PYP(H) and PYP(L), indicating that the proton at Glu46 is not transferred before formation of PYP(M). Free trans-p-coumaric acid had a 1294-cm(-1) band, which was shifted to 1288 cm(-1) in the cis form. All the difference FTIR spectra obtained had the pair of bands corresponding to them, indicating that all the intermediates have the chromophore in the cis configuration. The characteristic vibrational modes at 1020-960 cm(-1) distinguished the intermediates. Because these modes were shifted by deuterium-labeling at the ethylene bond of the chromophore while labeling at the phenol part had no effect, they were attributed to the ethylene bond region. Hence, structural differences among the intermediates are present in this region. Bands at about 1730 cm(-1), which show that Glu46 is protonated, were observed for all intermediates except for PYP(M). Because the frequency of this mode was constant in PYP(B), PYP(H), and PYP(L), the environment of Glu46 is conserved in these intermediates. The photocycle of PYP would therefore proceed by changing the structure of the twisted ethylene bond of the chromophore.

Antibacterial action of tryptanthrin and kaempferol, isolated from the indigo plant (Polygonum tinctorium Lour.), against Helicobacter pylori-infected Mongolian gerbils.

We evaluated the effect of tryptanthrin and kaempferol, both isolated from Polygonum tinctorium Lour., against Helicobacter pylori colony formation in vitro and in H. pylori-infected Mongolian gerbils. H. pylori suspension was mixed with solution of tryptanthrin and/or kaempferol and placed onto agar plates. These plates were incubated at 37 degrees C, under 10% CO2 for 5 days, and the H. pylori colonies were counted. For the in vivo experiment, Mongolian gerbils were inoculated with H. pylori ATCC 43504 orally. After 4 weeks, the infected gerbils were given tryptanthrin and/or kaempferol, administered orally, twice a day for 10 days. The animals were killed and the number of live H. pylori in their stomachs was determined. In vitro both tryptanthrin and kaempferol significantly decreased the numbers of H. pylori colonies a dose-dependent manner. An additive effect on colony formation was observed with the combined use. In the in vivo experiment, oral administration of tryptanthrin and/or kaempferol significantly decreased the numbers of colonies in the gerbils' stomachs. We concluded that tryptanthrin and kaempferol were effective against H. pylori in vivo.

Pitfalls in the toxicological analysis of an isobutyl nitrite-adulterated coffee drink.

A forensic investigation was carried out on one poisoning case, where cyanide was first detected in an evidence sample of a canned coffee drink. A more complete study revealed that it had been adulterated with isobutyl nitrite (IBN) and not cyanide. We examined the detectivity of IBN and related compounds by headspace gas chromatography and capillary electrophoresis. IBN decomposes to isobutyl alcohol (iBuOH) and nitrite in aqueous solution, and under higher temperature and more acidic conditions, the rate of this reaction becomes more rapid. IBN was also produced by the esterification of iBuOH with nitrite below pH 5. Cyanide was produced in a coffee solution by the addition of nitrite below pH 6. An IBN-spiked canned coffee drink solution was stored at 4 degrees C and periodically analyzed for IBN, iBuOH, nitrite, nitrate, and cyanide. Since the IBN level decreased rapidly, iBuOH was produced in an almost 90% molar yield. Nitrite production reached a maximum of 40% molar recovery on the first day and then gradually disappeared. The nitrate level reached a plateau of approximately 60% molar recovery. Cyanide was also detected, and its level at the 14th day was approximately 0.26% molar recovery. These findings suggest that, in a coffee drink solution, IBN undergoes hydrolysis to produce iBuOH and nitric acid, which is oxidized to nitrate and also produces cyanide through the nonspecific oxidation of organic compounds under acidic conditions.

Extrahepatic large hepatocellular carcinoma with peritoneal dissemination: multimodal treatment, including four surgical operations.

We report a patient with extrahepatically growing large hepatocellular carcinoma (HCC) associated with disseminated intraabdominal tumor and spontaneous tumor bleeding who was treated with four operations, transcatheter arterial embolization, systemic chemotherapy, and hyperthermia. It took 12 months for the multimodal treatment to normalize the alpha-fetoprotein (AFP) level, and remission continued for 6 months. We performed the fourth surgical treatment for a recurrent abdominal tumor involving the small intestine and mesentery, but the patient died 26 months after the first admission. Multimodal treatment, including repeat surgical treatments, for such advanced HCC should be encouraged, to prolong life and to maintain quality of life.

Effects of perilla seed oil supplementation on leukotriene generation by leucocytes in patients with asthma associated with lipometabolism.

BACKGROUND: Dietary sources of alpha-linolenic acid, such as perilla seed oil, may have the capacity to inhibit the generation of leukotrienes (LTs) by leucocytes in patients with asthma, as has been reported with the consumption of other long-chain n-3 fatty acids. METHODS: The factors affecting the suppression of leukotriene (LT) C4 generation by leucocytes were examined by comparing the clinical features of patients with asthma who had been given dietary perilla seed oil (n-3 fatty acids). Group A consisted of patients in whom the leucocyte generation of LTC4 was suppressed by dietary perilla seed oil. Group B consisted of those in whom LTC4 generation was not suppressed. RESULTS: LTC4 generation by leucocytes decreased significantly in group A after 2 (p < 0.05) and 4 weeks (p < 0.05); conversely, it increased significantly in group B after 4 weeks (p < 0.05). The two study groups differed significantly in terms of LTC4 generation by leucocytes after 4 weeks of dietary supplementation (p < 0.05). Ventilatory parameters such as peak expiratory flow (PEF), forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV(1)) increased significantly after 4 weeks of dietary supplementation in group A (p < 0.05). Values of PEF, FVC, FEV(1) and maximum expiratory flow at 25% of the forced vital capacity (V(25)) differed significantly between groups A and B prior to dietary supplementation. Serum levels of total cholesterol, low-density lipoprotein (LDL) cholesterol and phospholipid were significantly decreased by dietary supplementation in group A after 4 weeks. Serum levels of total cholesterol, triglyceride, high-density lipoprotein cholesterol, LDL cholesterol and phospholipid differed significantly between the two study groups prior to dietary supplementation. Serum levels of triglyceride and LDL cholesterol differed significantly between the two study groups after 4 weeks of dietary supplementation. CONCLUSIONS: Dietary supplementation with perilla seed oil in selected patients with asthma suppresses the generation of LTC4 and is associated with clinical features such as respiratory function and lipometabolism.

New monoterpene glycoside esters and phenolic constituents of Paeoniae radix, and increase of water solubility of proanthocyanidins in the presence of paeoniflorin.

Seven new monoterpene glycoside esters related to paeoniflorin were isolated from Paeoniae Radix, together with polymeric proanthocyanidins, polygalloylglucoses and 48 known compounds (a benzoylsucrose, seven aromatic acids, adenosine, nine monoterpene glycosides, eight flavan-3-ols, a catechin dimer formed by oxidation, seven proanthocyanidins, three galloylsucroses, five galloylglucoses, and six ellagitannins). The structures of the new compounds were determined by spectral investigation including two-dimensional NMR techniques. In addition, increased water solubility of polymeric proanthocyanidin in the presence of paeoniflorin was examined by n-octanol-water partition and 1H-NMR spectral experiments.

[Thermoradiotherapy combined with daily administration of low dose cisplatin for locally advanced laryngeal carcinoma].

We applied thermoradiotherapy combined with daily administration of low dose cisplatin (CDDP) to five patients with locally advanced laryngeal carcinoma. The total response rate (CR + PR) was 100% after irradiation of 30 or 40 Gy. One case showed CR, and the larynx could be preserved by adding a full dose of irradiation. CDDP administration was discontinued in two cases due to renal dysfunction or thrombopenia, but other cases tolerated the therapy without severe side effects. Total laryngectomy and bilateral neck dissection was carried out in four cases after irradiation of 30 or 40 Gy. Removed larynxes were prepared for light microscopic observations of the serial sections. One of the four specimens revealed a CR histologically. The combination of thermoradiotherapy with low dose CDDP showed a remarkable effect on reduction of tumor size and cytotoxicity of tumor cells. This might contribute to saving the larynx of patients with locally advanced laryngeal carcinoma.

cDNA cloning and gene expression of phenylalanine ammonia-lyase in Lithospermum erythrorhizon.

Two cDNA clones (LEPAL-1 and LEPAL-2) encoding phenylalanine ammonia-lyase were isolated from cell suspension cultures of Lithospermum erythrorhizon. Northern kinetic studies showed that LEPAL-1 mRNA contents markedly increased one day after inoculation of the cells into fresh medium, then decreased to the steady-state level. The course of mRNA accumulation paralleled that of PAL enzyme activity. The rapid induction of PAL activity seems to reflect the induction of dihydroechinofuran biosynthesis, while shikonin was produced at the steady-state level of PAL activity. The course of LEPAL-2 mRNA accumulation seemed to be similar to, but much lower than that of LEPAL-1. In the intact plant, both genes are expressed mainly in the root, the organ in which shikonin is exclusively produced and accumulated. Genomic Southern blot analyses showed that both genes are present in the genome as single copies.

[Exposure to blood during midwifery operations--a questionnaire study].

A self-administered questionnaire was mailed to all midwives working in Nara Prefecture to investigate blood exposure events at delivery and to consider protective measures for the exposure. Out of the 203 midwives 193 responded to the questionnaire. The median number of deliveries in which the midwives had assisted within the past one year was 35 after excluding those who had not assisted in deliveries at all. The incidence of needle-stick injuries was estimated to be 86.2 per 100 persons within the past one year. The occurrence in the subgroup who had worked as midwife for less than 5 years was 2-3 times higher than that in those who worked for more than 5 years. Over 90% of the midwives had experiences of direct blood contact events to the skin while assisting in their 10 most recent deliveries. Blood contact events occurred in the finger-hand-arm area in 85% of the midwives, on the legs in 62%, in the face in 20% and in the eyes in 1%. Around 20% of the midwives had unintentionally sucked amniotic fluid when using a tracheal catheter on a newborn. The most common occasion where direct blood contact events occurred was while bathing an infant for removing blood and amniotic fluid. The midwives wore a disposal gown more often when assisting in labors of parturient women infected with blood-born infectious diseases than without the diseases, and similarly for wearing gloves when measuring blood loss and for usage of a mechanical device for sucking amniotic fluid in the tracheae of an infant. Based on the results obtained in this questionnaire study and our previous study about blood contact events observed in a delivery room, protective measures for midwives against exposure to blood at delivery are required and some ideas are presented.

[Exposure to blood during midwifery procedures--blood contact events to midwives occurred in a delivery room].

Midwives are often exposed to blood during delivery procedures. A study aimed at clarifying actual status of blood contact during midwifery procedures was performed. Observations in a delivery room were conducted to record blood contact events experienced by midwives from the time of episiotomy until 2 hour after expulsion of the placenta. All gloves used by midwives were collected and tested for holes. Blood contact was defined as any contact with blood of a parturition woman as recognized visually by an observer. During a one-month study period data was obtained from a total of 19 midwives (12 midwives and 7 student midwives) who assisted in 8 deliveries. All of the midwives wore gowns with long sleeves, gloves and caps but did not use eye protection or masks. None of the parturient women had Hepatitis B, Hepatitis C or acquired immunodeficiency syndrome. Deliveries caused widespread blood exposure to the midwives throughout the complete course. Protection from most of this exposure was provided by the gowns and gloves. However, direct blood contacts occurred to the fingers, hands or forearms in several midwives. These events were caused by partly because the midwives unintentionally performed procedures without gloves and partly because blood penetrated the gown and soaked to the skin. Direct blood contacts to the foot in two midwives and to the mouth in one also occurred. The overall perforation rate for gloves examined was 4 out of 154 (2.6%). Two gloves were broken during washing sharp instruments contaminated with blood, one was torn when wearing, and the remaining one appeared to have had a hole prior to use.(ABSTRACT TRUNCATED AT 250 WORDS)

Scintigraphic study on the distribution of radiolabeled cis-diamminedichloroplatinum (II) in the tumor-bearing rabbit: a comparison between intra-arterial injection with lipiodol and intravenous injection.

To compare the distribution of cis-diamminedichloroplatinum (II) (CDDP) on scintigraphic images between intra-arterial injection with lipiodol and intravenous injection, we obtained serial scintigraphic images using radiolabeled CDDP (Pt-195m CDDP), which had a high specific activity (7.4 x 10(6) Bq [200 microCi]/mg-CDDP), in the rabbit VX-2 tumor system. A dose of 9.25 x 10(6) Bq (250 microCi) of Pt-195m CDDP was injected at one shot via an ear vein, while 7.4 x 10(6) Bq (200 microCi of Pt-195m CDDP mixed with 1 ml of lipiodol was injected at one shot via a saphenous artery. Mean pixel count, which was corrected for the total dose injected, in the arterially injected tumor always exceeded than that in the intravenously injected tumor. Pt-195m CDDP uptake by the rabbit tumor at 2 hr was 7.8% of the total injected dose with intravenous injection and 16.0% with intra-arterial injection. These data suggest that intra-arterial injection with lipiodol is a more useful method to increase CDDP uptake by tumors than intra-venous injection.

[Effects of MRX-III on chronic renal failure in rats].

The effects of MRX-III, a new amino acid solution for renal failure, on survival, progression of renal insufficiency, metabolism of calcium and phosphorous, and nutritional status were studied in rats with chronic renal failure induced by 7/8 renal ablation. These rats were injected intraperitoneally with MRX-III, an essential amino acid solution for renal failure (Amiyu; Sol. A) or a general amino acid solution (Moripron-F; Sol. M) for 12 weeks under the condition of a 3.5% protein diet, and these effects were compared with those in control rats infused with Sol. M under a 22% protein diet.1) Infusion of MRX-III or the other solutions under a low protein diet prolonged survival time and improved the uremic status indicated by azotemia, polyuria, albuminuria, hypocalcemia and hypertension. 2) Increase in body weight and tissue weight of rats treated with MRX-III or Sol. A was better than those in rats treated with Sol. M. MRX-III as well as Sol. A showed a tendency to provide a better nutritional effect in comparison with Sol. M.

[Effects of an essential trace element agent (TE-5) for total parenteral nutrition on the mineral nutrition in rats fed a trace element-deficient diet].

TE-5 is an essential trace element agent containing iron, zinc, copper, manganese and iodine for total parenteral nutrition (TPN). We have already reported that TE-5 improved the reduction of trace element concentrations induced by TPN. However, effects of TE-5 on the changes in biological function relating to trace elements are poorly understood. The present study was designed to clarify the effects of TE-5 on these functions. Rats fed a trace element (iron, zinc, copper, manganese and iodine)-deficient diet for 7 weeks showed reductions in the following parameters: plasma and various tissue concentrations of iron, zinc, copper, manganese and iodine, growth rate, erythrocyte (iron), hemoglobin (iron), hematocrit (iron), mean corpuscular constants (iron), plasma alkaline phosphatase activity (zinc), serum ceruloplasmin concentration (copper), liver pyruvate carboxylase activity (manganese) and serum thyroxine concentration (iodine). On the other hand, when TE-5 (0.008, 0.04 and 0.2ml/kg: x 0.2, x 1 and x 5 the usual clinical dose, respectively) was intravenously administered once a day for 7 weeks under the conditions described above, there was a tendency to prevent the reductions of plasma and various tissue concentrations of iron, zinc and manganese. In addition, TE-5 prevented the reductions of growth rate, iron metabolism functions, plasma alkaline phosphatase activity, serum ceruloplasmin concentration and liver pyruvate carboxylase activity. The present study shows that TE-5 prevents both reductions of trace element contents and trace element-related functions, and suggests that TE-5 is useful for treatment of trace element deficiency in TPN.

[Temperature distribution and geometry of the electrodes in RF interstitial hyperthermia using circular and interstitial electrodes].

To evaluate the feasibility of clinical application of a newly developed interstitial hyperthermia system, which consists of an 8 MHz radiofrequency generator, interstitial needle electrodes, and a superficial circular electrode, we conducted preclinical experiments using an agar phantom and VX-2 carcinoma in the rabbit. In the experiment with an agar phantom, four 4 cm needle electrodes were placed in a square array at intervals of 1.0, 1.5, and 2.0 cm. Thermography demonstrated homogeneous temperature distribution at electrode intervals of 1.0 and 1.5 cm, but hot spots around the electrodes at an interval of 2.0 cm. When electrode deviation was less than 8 degrees from the parallel plane, no temperature deviation was observed. Using two 2 cm electrodes and two 4 cm electrodes in square array, thermography demonstrated a homogeneous temperature distribution in the area surrounded by the electrodes. Even if the electrodes were located at the periphery of the agar phantom, a homogeneous temperature distribution was obtained in the area surrounded by the electrodes. Using four 4 cm electrodes at intervals of 1.5 cm in VX-2 carcinoma in the rabbit, ideal heating was obtained: 42 degrees C at the periphery of the tumor and 43 degrees C at the center. These data suggest that the newly developed interstitial hyperthermia apparatus provides homogeneous heat distribution at electrode intervals of 1.5 cm or less and can be used in a Phase I study for deep-seated or superficial tumors.

[Combined treatment in patients with carcinoma in situ of the urinary bladder using intravesical pirarubicin, irradiation and hyperthermia].

The treatment with hyperthermia in combination with radiation and intravesical pirarubicin (THP-ADM) was preliminary investigated in 5 patients with urinary bladder carcinoma in situ. Following intravesical administration of 30 mg THP-ADM, external irradiation of 3.0 Gy was delivered to the urinary bladder. Immediately then, hyperthermia using Thermotron RF-8 was performed for 50 min (intravesical temperature: 42-43 degrees C for 35 min). After five courses of the treatment, complete response has been maintained for 6, 8, and 9 months in 3 patients. However, in a patient with complete response, urinary cytology became positive in the 6th month after the treatment. In the remaining patient treatment was interrupted after only 3 courses due to urinary irritation, urinary cytology didn't become negative. The side effects of the combined treatment were limited to the transient symptoms of bladder irritation in all patients and thermal burn in 2 patients. These preliminary results suggest that this combined treatment may represent an effective conservative therapy for patients with urinary bladder carcinoma in situ.

[Supplementation of essential trace elements during total parenteral nutrition--effects on trace element-deficient rats].

Thirty-one male SD rats, six weeks old, were fed a trace element-deficient diet for two weeks and then divided into three groups and maintained for 1 week as follows: group A with total parenteral nutrition (TPN) without supplementation of trace elements, group B with TPN supplemented with the following 5 trace elements ... iron, zinc, copper, manganese and iodine, and group C with a diet free of the above five trace elements. Another group of eight rats was fed a diet supplemented with the above five trace elements for three weeks as a control (group D). Feeding or TPN without supplementation of trace elements evoked microcytic hypochromic anemia and significant decreases in iron concentrations in plasma and tissues (groups A and C). Supplementation of trace elements in the TPN solution showed a tendency to cure anemia and a significant increase in the iron concentration in tibia (group B). Decreases in the zinc or copper concentrations in plasma and tissues during TPN without trace elements were prevented by supplementation of trace elements in the TPN solution (group B). The plasma zinc and copper concentrations correlated well with their levels in liver, kidney and tibia. Manganese deficiency was not recognized in this investigation (groups A and C), though supplementation of trace elements in the TPN solution increased tissue manganese concentration (group B). Feeding or TPN without supplementation of trace elements induced decreases in plasma triiodothyronine and thyroxine (groups A and C). Supplementation of trace elements in the TPN solution showed a tendency to increase plasma thyroxine (group B).(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of total parenteral nutrition containing essential trace elements on their concentrations in rats].

Twenty-four rats were equally divided into 4 groups and maintained for 1 week as follows: Group A (normal control) with a synthetic normal diet and distilled water, group B with conventional total parenteral nutrition (TPN), i.e., TPN without essential trace elements (ETE), group C with TPN supplemented with a usual dose of ETE solution (TE-5), and group D with TPN supplemented with 3 times the usual dose of TE-5. Body weight, trace element concentrations in various tissues and certain blood biochemical parameters were determined in these rats. The results were as follows: 1) No significant differences in body weight were observed among the groups. 2) The iron concentrations in plasma and tibia decreased significantly in group B as compared with group A. The addition of TE-5 prevented these decreases, but dose-dependent increases in the concentrations of iron were observed in liver, spleen and kidney (groups C and D). 3) The zinc concentrations in plasma, whole blood, brain, heart, kidney and tibia decreased significantly in group B as compared with group A. The addition of TE-5 prevented these decreases dose-dependently (groups C and D). 4) The copper concentrations in plasma, whole blood, liver, spleen, kidney, testis and tibia decreased significantly in group B as compared with group A. The addition of TE-5 resulted in a tendency for these decreases to diminish (groups C and D). 5) The manganese concentrations of whole blood in group B decreased significantly as compared with group A. The addition of TE-5 caused the manganese concentrations of various tissues in groups C and D to increase significantly as compared with group A.(ABSTRACT TRUNCATED AT 250 WORDS)