Kan Usuda

Urinary biomarkers monitoring for experimental fluoride nephrotoxicity

Abstract An excess of sodium fluoride (135 mg F/kg body weight) was given in a single oral dose to male Wistar rats. Effects were investigated of fluoride-induced acute kidney intoxication on the time-dependent variations of urine volume. Also, of urinary fluoride ion (F−), α-glutathione-S-transferase (α-GST), N-acetyl-β-d-glucosaminidase (NAG), and creatinine (CR) concentrations. Fluoride administration strongly affects these urinary biochemical indices. Of the several biomarkers studied, α-GST is particularly useful as marker of S3 proximal tubule damage. We found that α-GST shows the strongest and more durable changes as a result of the large dose of F− given to the experimental animals. Our results suggest that the toxic effect of F− on the kidney may be more pronounced in the proximal tubule than the glomeruli region, and that the disorder of the proximal tubule is more serious in the S3 segment than S1 or S2 segment. α-GST proved to be a useful marker for the early detection and long-term observation of proximal renal tubular injury resulting from F− intoxication. The animal model should help to establish guidelines for the treatment of industrial workers suffering from acute renal failure resulting from accidental exposure to fluoride.

Successful treatments of lung injury and skin burn due to hydrofluoric acid exposure

Abstract Recent growth in the electronics and chemical industries has brought about a progressive increase in the use of hydrofluoric acid (HF), along with the concomitant risk of acute poisoning among HF workers. We report severe cases of inhalation exposure and skin injury which were successfully treated by administering a 5% calcium gluconate solution with a nebulizer and applying 2.5% calcium gluconate jelly, respectively. Case 1: A 52-year old worker used HF for surface treatment after welding stainless steel, and was hospitalized with rapid onset of severe dyspnea. On admission to the critical care medical center he had widespread wheezing and crackles in his lungs. Chest radiograph showed a fine diffuse veiling over both lower pulmonary fields. Severe hypocalcemia with high concentrations of F in serum and urine were disclosed. He was immediately given 5% calcium gluconate solution by intermittent positive-pressure breathing (IPPB), utilizing a neblizer. On the 21st hospital day, chest film and CT scan did not demonstrate any abnormality. He was discharged very much improved on the 22nd hospital day. Case 2: A 35-year old worker at an electronics factory was admitted to his local hospital with severe skin burn on his face and neck after exposure to 100% HF. Treatment began with immediate copious washing with water for 20 min. Calcium gluconate 2.5% gel (HF burn jelly) was applied to the area as a first-aid measure. Persistent high concentrations of serum and urinary F were disclosed for 2 weeks. After treatment with applications of HF burn jelly, he was confirmed as being completely recovered. The present cases and a review of published data suggest that an adequate method of emergency treatment for accidental HF poisoning is necessary.

Serum and urinary boron levels in rats after single administration of sodium tetraborate

Abstract The pharmacokinetics of boron was studied in rats by administering a 1 ml oral dose of sodium tetraborate solution to several groups of rats (n=20) at eleven different dose levels ranging from 0 to 0.4 mg/100 g body weight as boron. Twenty-four-hour urine samples were collected after boron administration. After 24 h the average urinary recovery rate for this element was 99.6 ± 7.9. The relationship between boron dose and excretion was linear (r=0.999) with a regression coefficient of 0.954. This result suggests that the oral bioavailability (F) of boron was complete. Another group of rats (n=10) was given a single oral injection of 2 ml of sodium tetraborate solution containing 0.4 mg of boron/100 g body wt. The serum decay of boron was followed and found to be monophasic. The data were interpreted according to a one-compartment open model. The appropriate pharmacokinetic parameters were estimated as follows: absorption half-life, t1/2a=0.608±0.432 h; elimination half-life, t1/2=4.64±1.19 h; volume of distribution, Vd=142.0±30.2 ml/100 g body wt.; total clearance, Ctot=0.359 ± 0.0285 ml/min per 100 g body wt. The maximum boron concentration in serum after administration (Cmax) was 2.13 ± 0.270 mg/l, and the time needed to reach this maximum concentration (Tmax) was 1.76 ± 0.887 h. Our results suggest that orally administered boric acid is rapidly and completely absorbed from the gastrointestinal tract into the blood stream. Boric acid in the intravascular space does not have a strong affinity to serum proteins, and rapidly diffuses to the extravascular space in proportion to blood flow without massive accumulation or binding in tissues. The main route of boron excretion from the body is via glomerular filtration. It may be inferred that there is partial tubular resorption at low plasma levels. The animal model is proposed as a useful tool to approach the problem of environmental or industrial exposure to boron or in cases of accidental acute boron intoxication.

An Overview of Boron, Lithium, and Strontium in Human Health and Profiles of These Elements in Urine of Japanese

The biological, medical and environmental roles of trace elements have attracted considerable attention over the years. In spite of their relevance in nutritional, occupational and toxicological aspects, there is still a lack of consistent and reliable measurement techniques and reliable information on reference values. In this review our understandings of the urinary profilings of boron, lithium and strontium are summarized and fundamental results obtained in our laboratory are discussed.Over the past decade we have successfully used inductively coupled plasma emission spectrometry for the determination of reference values for urinary concentrations of boron, lithium and strontium. Taking into account the short biological half-life of these elements and the fact that their major excretion route is via the kidney, urine was considered to be a suitable material for monitoring of exposure to these elements. We confirmed that urinary concentrations of boron, lithium and strontium follow a lognormal distribution. The geometric mean reference values and 95% confidence intervals were 798 μg/l (398–1599 μg/l) for boron, 23.5 μg/l (11.0–50.5 μg/l) for lithium and 143.9 μg/l (40.9–505.8 μg/l) for strontium. There were no discrepancies between our values and those previously reported. Our reference values and confidential intervals can be used as guidelines for the health screening of Japanese individuals to evaluate environmental or occupational exposure to these elements.

Toxicokinetics of intravenous fluoride in rats with renal damage caused by high-dose fluoride exposure

Abstract Fluoride (F) complexes are used in some fields of industry and medicine. F excretion mainly depends on kidney function. Urinary F concentration is measured to monitor the health of workers exposed to F. The toxicokinetics of F were studied by analyzing plasma concentration of F after intravenous injection of 2.86, 5.71 and 8.57 mg/kg into male Wistar rats. A dose–response relationship was recognized between these F doses and renal tissue injury. Blood samples were removed at 0, 10, 20, and 30 min, and after 1, 2, 3, 4, 5, and 6 h after injection. Plasma concentration-vs-time profiles were evaluated by a nonlinear least-squares method for fitting data to polyexponential equations and calculation of relevant pharmacokinetic parameters. Results indicated that a two–compartment model could describe the elimination of F from plasma. The β rate constant, total plasma clearance (Cl) and first-order rate constants (K21, Kel) decreased, and the half-time of the β-phase (t1/2β) was significantly prolonged with increasing dose. The kidney is the main target organ for F toxicity. Acute exposure to high doses of F damages renal tissue and causes renal dysfunction. The Cl of F is mainly dependent on renal F excretion. Since severe kidney damage markedly affected the toxicokinetics of F and decreased its elimination, other nephrotoxic indicators and measurement of plasma F concentration are necessary for monitoring high-dose F exposure.

Determination of reference concentrations of strontium in urine by inductively coupled plasma atomic emission spectrometry.

ObjectiveThe aim of this study was to establish reference concentrations of urinary strontium by inductively coupled plasma atomic emission spectrometry (ICP-AES).MethodsFor the determination of strontium, urine samples were collected from healthy Japanese (n=146; 115 males, 31 females; mean age, 33±9 years; age range, 18 to 58 years). The urine samples stored at or below −20°C were thawed with incubation at 40°C for 30 min and sediments were dissolved by vigorous shakings. Then, the samples were centrifuged at 3000 g for 5 min, and the supernatant was directly aspired into a P-5200-3600/1200 ICP-AES system from Hitachi Ltd., Tokyo, Japan.ResultsA steeper increase in the S/N ratio and a good effective linearity of the calibration line was obtained at 407.771 nm in the range of 0–300 μg/L strontium standard solution. Urine samples having the same background signal as that of 18 MΩ cm ultrapure blank water, a good correspondence of the single peak pattern of the spectra, accuracy and precision of spike recovery were also confirmed. Urinary strontium concentrations showed a log-normal distribution and a geometric mean concentration of 143.9 μg/L, with 5–95% confidential interval of 40.9–505.8 μg/L.ConclusionThe results of this study will be useful as guidelines for the biological monitoring of strontium in normal subjects and in individuals therapeutically or environmentally exposed to strontium.

Serum boron concentration from inhabitants of an urban area in Japan: Reference value and interval for the health screening of boron exposure

Boron (B) levels were determined in the serum of 980 healthy inhabitants living in an urban area of Japan by means of inductively coupled plasma emission spectrometry (ICPES).The results showed a log-normal distribution of serum B for both sexes, although there are age-related differences. In male subjects, serum B increases rapidly up to 49 yr of age, reaching a plateau between ages 50 and 69 yr old, followed by a gradual increase up to 70 yr or older. Female subjects exhibit a gradual increase up to the age of 70 yr old. The reference value for male and female subjects was 79.8 μg/L and 67.9 μg/L, and the reference interval was 33.3–191.2 μg/L and 29.5–154.9 μg/L, respectively.The obtained reference value and interval of the nonexposed group may be useful for health screening for B exposure, either for people living in regions with high levels of B in the environment, or for workers who are exposed to this element.

Study on urine boron reference values of Japanese men: Use of confidence intervals as an indicator of exposure to boron compounds

A simple and rapid method for the determination of urine boron by inductively-coupled plasma argon emission spectrometry (ICPAES) has been developed to establish boron exposure guidelines. After 11-fold dilution in 18.25 M omega cm ultra-pure water and vigorous shaking, urine may be directly injected into the spectrometer, providing accurate and reproducible results. We report the results obtained with urine samples obtained from a group of male Japanese electronic workers (n = 102) who had not been exposed to boron; boron concentrations were corrected with use of a specific gravity of 1.024 g/ml. The frequency distribution resulted in a log-normal distribution diagram for anatomical spread. The geometric mean values for urine boron in the non-exposed workers was 798.0 micrograms/l, while the confidence interval (C.I.) was between 398.1 and 1599.6 micrograms/l. Taking into consideration the short biological half-life of boron and its major excretion route via urine, urine was considered to be a suitable means for monitoring of exposure to this element. We conclude that the guidelines established by determining boron reference values are useful for the protection of individuals exposed to boron in their working environments.

Systemic effects and skin injury after experimental dermal exposure to monochloroacetic acid

There have been many fatal occupational accidents of skin exposure to monochloroacetic acid (MCA). However, there have been no reports of dermatological findings and the lethal consequences have not yet been demonstrated. Therefore, harmful local and systemic effects were investigated after dermal exposure to MCA. A 0.5 mL aliquot of MCA solution (40% w/w) was applied to the abdominal skin of ten 10-week-old male SD rats under anesthesia. The exposure area (25 × 25 mm2) was 1.6% of the total surface area. The dose of MCA per area was 34.1 mg/cm2. Saline was similarly administered to 10 control rats. Histopathological findings after 10 min were observed by light microscopy. Blood samples were collected by exsanguinations from the carotid arteries after 4 h. Skin samples were collected 10 min after the initial exposure. Histological findings showed severe degeneration of collagen bundles in the epidermis and subcutaneous tissues. PCO2, HCO3 −, TCO2, BE and glucose levels were decreased in the MCA group. AST, m-AST, ALT, BUN, Cr, NH3, lactic acid, pyruvic acid, RBC, Hb, Hct, total protein and albumin were increased in the MCA group. The burn was determined to be a third-degree burn on the basis of the histopathological findings. The severe toxicity was probably a consequence of the rapid permeability. Biochemical parameters were a consequence of hepatocellular injuries, renal dysfunction, dysglyconeogenesis and dysfunction of ammonia metabolism. MCA reportedly enters the TCA cycle and inhibits aconitase. MCA metabolites also inhibit pyruvate carboxylase in the gluconeogenesis pathway. Therefore, the important serum biochemical abnormalities such as hypoglycemia and lactic acidosis should be monitored to find the acute systemic disorders.

Markers of cadmium exposure in workers in a cadmium pigment factory after changes in the exposure conditions

The objective of this study was to assess changes in concentrations of cadmium in the blood (Cd-B), cadmium in the urine (Cd-U), b2-microglobulin in the serum (b2-mG-S) and b2-microglobulin in the urine (b2-mG-U) of workers at a cadmium (Cd) pigment factory in Japan in which exposure conditions improved. We evaluated reversibility of these markers in continuously employed workers in relation to changes in exposure levels resulting from improvements in the workplace and the reduced production of Cd. Our study involved both environmental and biological monitoring. Data were collected for four years. We measured the Cd concentration in the air of each work area, using the time-weighted average (TWA). Cd-B and Cd-U were measured in workers as direct indices of Cd exposure. b2-mG-S and b2-mG-U were measured as markers of renal tubular function. Exposure levels were high in all work areas, according to the criteria set by the American Conference of Governmental Industrial Hygienists (ACGIH). Workers’ Cd-B and Cd-U concentrations reflected high levels of exposure. Correlation was found between these direct indices and b2-mG-S concentrations. Since the second year, ambient Cd concentrations decreased and reacted markers have been improved. Our results suggest that Cd-B, Cd-U, b2-mG-S and b2-mG-U are appropriate markers for monitoring both the level of Cd exposure and the tubular function of workers. Reversibility of urinary low molecular weight protein was observed in the workers over the four years.