Temperature dependence on the pesticide sampling rate of polar organic chemical integrative samplers (POCIS).

Laboratory experiments were performed to determine the sampling rates of pesticides for the polar organic chemical integrative samplers (POCIS) used in Japan. The concentrations of pesticides in aquatic environments were estimated from the accumulated amounts of pesticide on POCIS, and the effect of water temperature on the pesticide sampling rates was evaluated. The sampling rates of 48 pesticides at 18, 24, and 30 °C were obtained, and this study confirmed that increasing trend of sampling rates was resulted with increasing water temperature for many pesticides.

The subjective assessment of work and social adjustment impairments and associated psychopathologies in Japanese adult female patients with anorexia nervosa.

Aim: Patients with anorexia nervosa (AN) sometimes undergo a chronic course, and they hardly maintain social participation. Work and social adjustment impairments are generally significantly associated with the clinical symptoms of eating disorders. Psychopathologies associated with the subjective social difficulties of patients with AN have been unclear. This study examined the association between AN psychopathologies and work and social adjustment impairments in adult female patients with AN. Methods: This study included 36 Japanese adult female patients with AN who completed the Work and Social Adjustment Scale (WSAS) and the Eating Disorder Inventory-2 (EDI-2). Spearman's rank correlation coefficient was used to assess correlations between WSAS and EDI-2 or demographic variables. Results: The mean age was 31.8 years, the mean current body mass index was 13.4 kg/m2, and the median illness duration was 5 years. Patients demonstrated social difficulties, especially in social leisure activities. The total WSAS scores were significantly correlated with EDI-2 "impulse regulation" and "asceticism." WSAS "social leisure" was significantly correlated with EDI-2 "bulimia," "interoceptive awareness," "impulse regulation," and "asceticism." Conclusion: Psychopathologies, such as impulse regulation, asceticism, and interoceptive awareness, may be related factors to social difficulties. Emotion regulation, such as impulse regulation and emotional awareness, could be an important realm of treatment not only for psychopathology but also for social functioning in patients with AN.

Development of a New Polar Organic Chemical Integrative Sampler for 1,4-dioxane Using Silicone Membrane as a Diffusion Barrier.

Efficient monitoring methods must be developed for 1,4-dioxane, which is suspected to be carcinogenic to humans and is highly mobile in aquatic environments. In this regard, polar organic chemical integrative samplers (POCIS) have been utilized extensively as passive samplers for determining time-weighted average concentrations of hydrophilic organic compounds. However, POCIS are difficult to apply to extremely hydrophilic known organic compounds with negative log octanol-water partition coefficient (Kow ) values due to their limited kinetic sampling time. Using an activated carbon-based sorbent with a high adsorption capacity and a bilayer of silicone and polyethersulfone membranes that inhibit mass transfer to the sorbent, we developed a POCIS device to measure 1,4-dioxane (log Kow -0.27) in the present study. Permeation and field calibration tests demonstrated that the use of silicone membranes effectively reduces the water-to-sorbent mass transfer rate. The sampling rate and kinetic sampling period determined by field calibration tests were 1.4 ml day-1 and >14 days, respectively. Finally, the developed POCIS device was applied to a landfill treatment plant to determine the 1,4-dioxane concentrations. Environ Toxicol Chem 2023;42:296-302. © 2022 SETAC.

Valpha14 NK T cell-triggered IFN-gamma production by Gr-1+CD11b+ cells mediates early graft loss of syngeneic transplanted islets.

Pancreatic islet transplantation is a highly promising approach for the treatment of insulin-dependent diabetes mellitus. However, the procedure remains experimental for several reasons, including its low efficiency caused by the early graft loss of transplanted islets. We demonstrate that Gr-1+CD11b+ cells generated by transplantation and their IFN-gamma production triggered by Valpha14 NKT cells are an essential component and a major cause of early graft loss of pancreatic islet transplants. Gr-1+CD11b+ cells from Valpha14 NKT cell-deficient (Jalpha281-/-) mice failed to produce IFN-gamma, resulting in efficient islet graft acceptance. Early graft loss was successfully prevented through the repeated administration of alpha-galactosylceramide, a specific ligand for Valpha14 NKT cells, resulting in dramatically reduced IFN-gamma production by Gr-1+CD11b+ cells, as well as Valpha14 NKT cells. Our study elucidates, for the first time, the crucial role of Gr-1+CD11b+ cells and the IFN-gamma they produce in islet graft rejection and suggests a novel approach to improving transplantation efficiency through the modulation of Valpha14 NKT cell function.

Comparative Evaluation of the Polar Organic Chemical Integrative Sampler in Two Types of Validation Systems Simulating Peak Concentration Events.

Polar organic chemical integrative sampler (POCIS) devices have been suggested for measuring time-weighted averages (TWAs) of contaminant concentrations resulting from chemical leak accidents in aquatic environments. However, the response of the POCIS device in the emergency condition in natural water remains unclear. The response of the POCIS device to contaminant fluctuation was investigated using a chamber test with tap water and a channel test with natural water. The fluctuation in the chamber and the channel simulated the condition of river water under a chemical leak scenario (maximum concentration: 1-10 µg L-1 , half-life: 1 day). The target chemicals were neonicotinoid pesticides (dinotefuran, clothianidin, thiamethoxam, imidacloprid, acetamiprid, and thiacloprid) and bisphenol A. The ratio of the POCIS measured value to the TWA values of grab samplings (POCIS/TWA) for the channel test (temperature: 15 °C, flow velocity: 15 cm s-1 ) ranged from 61% (clothianidin) to 133% (thiacloprid). The results indicated that the POCIS device could be effectively used as a monitoring device in an aquatic environment under the chemical leak scenario over a time period of more than14 days. In addition, the POCIS/TWA ratios obtained from the chamber test and the channel test were in the range of 50-150%. Thus, the chamber test could be used to evaluate the POCIS device at a low cost. Environ Toxicol Chem 2021;40:3010-3018. © 2021 SETAC.

Serum γ-glutamyltransferase, triglyceride and total cholesterol are possible prediabetic risk markers in young Japanese men.

Serum profiles of lipids and/or liver enzymes are established markers for the estimation of insulin resistance and diabetic risk in the non-diabetic middle-aged population. To identify prediabetic markers in young subjects, 110 young male subjects (20-29 years of age) with normal glucose tolerance (NGT) were divided into two groups by median body mass index (BMI), <22.18 (n=55) and ≥22.18 (n=55) kg/m(2). Indices of insulin sensitivity including HOMA-IR and ISI composite, indices of ß-cell function including HOMA-ß, insulinogenic index (ΔI(30)/ΔG(30)) and ΔI(30)/ΔG(30)/ HOMA-IR were calculated. Statistical associations between these parameters and the serum lipid profiles and liver function were evaluated. Alanine aminotransferase (ALT), γ-glutamyltransferase (GGT), total cholesterol (TC) and triglyceride (TG) levels were inversely correlated with the ISI composite among individuals with BMI ≥22.18 kg/m(2) but not those with BMI <22.18 kg/m(2). Multivariate regression analysis revealed that, in Group N, the plasma glucose levels at 60 min (PG(60)) were inversely correlated with the ISI composite and the insulinogenic index, and were positively correlated with the GGT, TC and TG levels. On the other hand, in Group L, PG(60) was correlated with the insulinogenic index, TC and TG levels. In conclusion, elevated levels of GGT, TC and TG are good clinical markers to predict diabetic risks, even in young NGT males. Of these, GGT was the most strongly related factor among subjects with relatively high BMI.

Acute inflammatory syndrome and intrahepatic cholestasis caused by an interleukin-6-producing pheochromocytoma with pregnancy.

We herein describe the case of a 30-year-old woman who experienced high fever during the puerperal period and was diagnosed with pheochromocytoma. Acute inflammatory syndrome, as indicated by the elevated serum levels of interleukin-6 (IL-6), and cholestatic liver dysfunction were observed. Since this condition resolved before the operation, it was probably caused by massive central necrosis within the tumor. The IL-6 production from the tumor cells was confirmed by immunohistochemistry. When a case of pheochromocytoma accompanied with acute inflammatory syndrome is encountered, the possibility that the tumor itself might produce some cytokines should be considered, even in the presence of massive necrosis within the tumor.

[Case of enzyme-linked immunoglobulin with persistent elevation of lactate dehydrogenase activity in serum by administration of an anti-psychotic drug].

We presented a case, who showed extremely high activity of lactate dehydogenase (LD) and confirmed the presence of the LD linked immunoglobulin in her serum. The maximum activity of LD was 6830 IU/L, and the electrophoretic pattern of LD isozymes showed the broad spectrum from isozyme LD2 to LD5. Analysis by counter immuno electrophoresis revealed that immunogloblin was attached to the M subunit of LD and its subtype was an IgG, lambda-chain. The cause, which produced the complex, might be thought to be the side effects by tiapride administrated for her mild dementia. After discontinuance of this drug, the LD activity in serum had gradually reduced. The serum creatinine also increased gradually after administration of tiapride, and did not reverse to normal level by discontinuance of it. The patient died from acute renal failure, which aggravated from sporadic urinary tract infection. It was suggested that her basal renal dysfunction might be due to the LD IgG complexes. We propose a rapid disruption of suspicious drug for the course of the production of LD linked immunoglobulin, because very high titer of these complexes might suffer irreversible damage to the kidney, which chance to become acute renal failure.

Determining the suitability of a polar organic chemical integrated sampler (POCIS) for the detection of pesticide residue in the Ishikawa River and its tributary in Osaka, Japan.

The monitoring of pesticide concentrations in Japanese rivers was conducted via a grab sampling method and a passive sampling method using the polar organic chemical integrated sampler (POCIS). The results showed that cumulative detections were 84 with grab sampling and 98 with the POCIS. All of the pesticides detected by grab sampling could be quantified with a POCIS except for one (although its traces were detected). In addition, 15 detections quantified by POCIS were undetected by grab sampling. The average concentrations of pesticides detected by both the POCIS and grab sampling during the investigation period were compared. A good correlation was observed between the two methods with a slope of 1.00 and a coefficient of correlation (r) of 0.897 (n=79). Although high temporal variability was observed in the pesticide concentrations by grab sampling, the average pesticide concentrations obtained by the two methods showed similar values during the investigation period.

Interaction of defibrillation waveform with the time to defibrillation or the number of defibrillation attempts on survival from out-of-hospital cardiac arrest.

AIM: Early biphasic defibrillation is effective in out-of-hospital cardiac arrest (OHCA) cases. In the resuscitation of patients with OHCA, it is not clear how the defibrillation waveform interacts with the time to defibrillation to influence patient survival. The second, and any subsequent, shocks need to be administered by an on-line physician in Japan. Thus, we investigated the interaction between the defibrillation waveform and time to or the number of defibrillation on resuscitation outcomes. METHODS: This prospective observational study used data for all OHCAs that occurred between 2005 and 2014 in Japan. To investigate the interaction effect between the defibrillation waveform and the time to defibrillation or the number of defibrillations on the return to spontaneous circulation (ROSC), 1-month survival, and cerebral performance category (CPC) (1, 2), we assessed the modifying effects of the defibrillation waveform and the time to or the number of defibrillation on additive scale (i.e., the relative excessive risk due to interaction, RERI) and multiplicative scale (i.e., ratio of odds ratios (ORs)). RESULTS: In total, 71,566 cases met the inclusion criteria. For the measure of interaction between the defibrillation waveform and the time to defibrillation, ratio of ORs for ROSC was 0.84 (0.75-0.94), implying that the effect of time to first defibrillation on ROSC was negatively modified by defibrillation waveform. For the interaction between the defibrillation waveform and the number of defibrillations, RERI and ratio of ORs for CPC (1, 2) was -0.25 (-0.47 to -0.06) and 0.79 (0.67-0.93), respectively. It is implied that the effect of number of defibrillation on CPC (1, 2) was negatively modified by defibrillation waveform. CONCLUSIONS: An increased number of defibrillations was associated with a decreased ROSC in the case of biphasic and monophasic defibrillation, while an increased number of defibrillations was related to an increased 1-month survival rate and CPC (1, 2) only in the case of biphasic defibrillation. When two or more defibrillations were performed, a biphasic waveform was more effective in terms of long-term survival than a monophasic waveform.

A Novel Subcutaneous Site of Islet Transplantation Superior to the Liver.

BACKGROUND: Islet transplantation is an attractive treatment for patients with insulin-dependent diabetes mellitus, and currently, the liver is the favored transplantation site. However, an alternative site is desirable because of the low efficiency of hepatic transplantation, requiring 2 to 3 donors for a single recipient, and because the transplanted islets cannot be accessed or retrieved. METHODS: We developed a novel procedure of islet transplantation to the inguinal subcutaneous white adipose tissue (ISWAT) of mice and described functional and morphological characteristics of transplanted syngeneic islets. Also, it was determined whether islet allograft rejection in the ISWAT can be prevented by immunosuppressive agents. Furthermore, it was examined whether human islets function when grafted in this particular site of immune-deficient mice. RESULTS: In this site, transplanted islets are engrafted as clusters and function to reverse streptozotocin-induced diabetes in mice. Importantly, transplanted islets can be visualized by computed tomography and are easily retrievable, and allograft rejection is preventable by blockade of costimulatory signals. Of much importance, the efficiency of islet transplantation in this site is superior to the liver, in which hyperglycemia of diabetic recipient mice is ameliorated after transplantation of 200 syngeneic islets (the islet number yielded from 1 mouse pancreas) to the ISWAT but not to the liver. Furthermore, human islets transplanted in this particular site function to reverse diabetes in immune-deficient mice. CONCLUSIONS: Thus, the ISWAT is superior to the liver as the site of islet transplantation, which may lead to improved outcome of clinical islet transplantation.

Natural killer T-cells participate in rejection of islet allografts in the liver of mice.

A role of natural killer T (NKT) cells in transplant rejection remains unknown. Here, we determined whether NKT cells participate in rejection of islet allografts, using NKT cell-deficient mice. Survival of islet allografts in streptozotocin-induced diabetic CD1d(-/-) mice or Valpha14 NKT cell(-/-) mice was significantly prolonged without immunosuppression when grafted into the liver, but not beneath the kidney capsule, compared with wild-type mice. Acceptance of intrahepatic islet allografts was achieved in CD1d(-/-) mice by a subtherapeutic dose of rapamycin, which was abrogated in conjunction with the transfer of hepatic mononuclear cells from wild-type, but not from CD1d(-/-), mice at islet transplantation. The second islet grafts from a donor-specific, but not from a third-party, strain in CD1d(-/-) mice bearing functional islet allografts were accepted without immunosuppression at 120 days after the initial transplantation. These findings demonstrate that NKT cells play a significant role in rejection of islet allografts in the liver of mice, but that NKT cells are not essential for induction of donor-specific unresponsiveness in this model. The current study indicates that NKT cells might be considered as a target for intervention to prevent islet allograft rejection when the liver is the site of transplantation.

Successful islet transplantation to two recipients from a single donor by targeting proinflammatory cytokines in mice.

BACKGROUND: Currently, the inability to achieve successful islet transplantation from one donor to one recipient is a major obstacle facing clinical islet transplantation. We herein determined whether this limitation could be overcome by targeting pro-inflammatory cytokines with the prevention of immediate islet graft loss in association with engraftment in mice. METHODS: Isolated islets were grafted into the liver of streptozotocin-induced diabetic mice and the role of proinflammatory cytokines in the engraftment of islets was evaluated with the use of interferon (IFN)-gamma-/- mice and monoclonal antibodies against proinflammatory cytokines. RESULTS: Hyperglycemia in streptozotocin-induced diabetic mice receiving 200 syngenic islets, which were isolated from a single mouse pancreas, was ameliorated when IFN-gamma-/-, but not wild-type mice, were used as recipients. The treatment with anti-IFN-gamma antibody produced normoglycemia in diabetic wild-type mice receiving 200, but not 100 islets. However, when anti-tumor necrosis factor-alpha and anti-interleukin-1beta antibodies were administered in conjunction with anti-IFN-gamma antibody, wild-type diabetic mice receiving 100 islets became normoglycemic after transplantation. In addition, the favorable effect of the combined use of antibodies was similarly achieved in mice receiving islet allografts when rejection was prevented with anti-CD4 antibody treatment. CONCLUSIONS: These findings clearly demonstrate that successful islet transplantation from one donor to two recipients is feasible by targeting pro-inflammatory cytokines in mice, thus suggesting a potential application in clinical islet transplantation if similar mechanisms of islet graft loss could be mediated in humans.

Beta-cell function is a major contributor to oral glucose disposition in obese Japanese students.

Determinants of glucose intolerance were studied in 163 obese Japanese young adults, 18 to 21 years old (43 females,120 males), who underwent 75-g oral glucose tolerance testing. Type 2 diabetes was newly diagnosed in 2.9% (n = 4); impaired fasting glucose (IFG) in 5.1% (n = 7); and impaired glucose tolerance (IGT) in 10.9% (n = 15). A homeostasis model assessment of insulin resistance (HOMA-IR) was used to estimate insulin sensitivity; beta-cell function during the first 30 min of the test was measured and defined as the insulinogenic index. This index was adjusted for insulin sensitivity, since this affects both beta-cell function and glucose disposition (disposition index). The relationship between insulinogenic index and 1/HOMA-IR was not hyperbolic. However, the disposition index (DI) was useful for the estimation of beta-cell function with the correct confirmation about it validity using beta-cell function index (BI). The association between insulin sensitivity and beta-cell function to glucose disposal, as measured by the area under the glucose curve (AUCg), was examined in all subjects. Insulin sensitivity was significantly related to AUCg (log HOMA-IR; R (2) = 0.142, p<0.0001). On the other hand, an inverse curvilinear relationship was observed between beta-cell function and AUCg (log(Delta I/Delta G)/HOMA-IR, R (2) = 0.411, p<0.0001). Thus, impaired beta-cell function, when estimated as DI, was strongly associated with impaired glucose disposal. In conclusion, our study showed that both insulin sensitivity and impaired beta-cell function are associated with impaired glucose metabolism, and that beta-cell function may be more important in determining glucose disposal.

[The effects of age on insulin sensitivity and insulin secretion in Japanese subjects with normal glucose tolerance].

The prevalence of impaired glucose tolerance and type 2 diabetes increases with age. However, controversial results have been reported in regard to which has a greater influence on the deterioration of glucose tolerance with age, namely impaired insulin sensitivity or impaired insulin secretion. The conflicting results may arise mainly from differences in the evaluation of insulin secretion and insulin sensitivity, and from differences in the physical composition and the ethnicity of the study subjects. We therefore selected Japanese subjects, between 20 and 80 years of age, with normal glucose tolerance (NGT) and with a body mass index (BMI) below 25.0 kg/m2, and then examined the subject's insulin sensitivity based on the indices of a homeostasis model assessment-insulin resistance index (HOMA-IR) and ISI composite (ISI), and beta-cell function by these of HOMA-beta, AUC I/G(0-120), an insulinogenic index (deltaI30/deltaG30), and then (deltaI30/deltaG30)/HOMA-IR derived from a 75 g-oral glucose tolerance test (OGTT). The subjects were divided into the six subgroups according to sex and age, below age 30, between ages 30 and 49, and equal to and over age 50. Both HOMA-IR and the ISI showed no differences across the range of age and sex. HOMA-beta decreased with age, and AUC I/G(0-120) decreased in the elderly. No change was observed in deltaI30/deltaG30 across the age range in men, however deltaI30/deltaG30/HOMA-IR, the index of the early phase insulin secretion adjusted for insulin sensitivity, decreased with age in both men and women. These data indicated that aging itself had no effect on insulin sensitivity, while insulin secretion in both the early and late phase during the OGTT deteriorated with age even within the NGT subjects.

[Clinical trial and evaluation of a direct report system of panic values at Fukuoka University Hospital].

In order to support a faster and more informative clinical practice, we established the criteria for panic (critical) values regarding the blood concentrations of glucose, Na, K, Ca, inorganic phosphate (IP), Hb and number of platelets, and also created a system to report these values directly to the doctors in charge. We initiated this system in September 2003. In order to evaluate the availability of this system, we analyzed the clinical data during a one year period, based on the findings of patients showing panic values, mainly concerning the disease states and the correspondences by the doctors who were directly informed. We also carried out questionnaire surveys about the panic values and the new system for all of the doctors in our hospital (recovery rate: 84.3%). The total number of panic values reported was 113 and the mean percentage of the number of ordered examinations was 0.019%. After the report, 79 cases (69.9%) were examined again or treated, while 34 cases (30.1%) had already been treated or watched carefully at the time of the report. Malignant diseases were the main causes of increased panic values (38 cases), especially in the Na, K and blood glucose of patients. The next disease state, which appeared to demonstrate high rates, was chronic renal failure (16 cases), in the low K, high Ca, and low IP patients. Most of the cases of low Hb were caused from bleeding of the gastro-intestinal tract, with malignancies next. A blood infusion was performed for all of the cases with low Hb except for one. As a result of the questionnaire survey among the staff doctors, we confirmed that this system did indeed work efficiently, and 88% of the doctors who answered the questionnaires, were satisfied with the system. In conclusion, we established a new system, which made it possible for panic values to be directly reported to the doctor in charge and this system was then evaluated for its clinical usefulness.

Age × Gender Interaction Effect on Resuscitation Outcomes in Patients With Out-of-Hospital Cardiac Arrest.

Although an interaction between gender and age has been shown to influence resuscitation outcomes in patients with out-of-hospital cardiac arrest (OHCA), this interaction has not been investigated in Asian populations. In this prospective, observational study, data from all cases of OHCA in Japan between 2005 and 2012 were obtained from the Japanese National Registry. We determined the relative excess risk due to interaction and the ratio of odds ratios (ORs) to assess the interaction effect of gender and age on the incidence of return of spontaneous circulation (ROSC) before hospital arrival, 1-month survival, and neurologically intact survival 1 month after OHCA. Male gender was associated with decreased ROSC and lower 1-month survival rates in patients with OHCA of presumed cardiac origin. Older age was associated with lower 1-month and neurologically intact survival rates in male patients with OHCA of presumed cardiac and noncardiac origin and with increased ROSC in male patients with OHCA of presumed cardiac origin. The relative excess risk due to interaction for ROSC in patients with OHCA of presumed cardiac origin was statistically significant (OR 0.19, 95% confidence interval [CI] 0.06 to 0.32). The ratio of ORs for ROSC was statistically significant in patients with OHCA of presumed cardiac origin (OR 1.25, 95% CI 1.05 to 1.47) and of noncardiac origin (OR 0.40, 95% CI 0.17 to 0.92). In conclusion, the interaction effect between age and gender on ROSC was positive in OHCA cases of presumed cardiac origin and negative in those of noncardiac origin.

[Treatment of type 1 diabetics by transplantation of isolated pancreatic islets].

Type 1 diabetes is an autoimmune disease with selective destruction of insulin-producing pancreatic beta cells. Since insulin plays pivotal roles in energy homeostasis by transferring glucose into cells, type 1 diabetic patients can not survive without insulin replacement. Insulin secretion is precisely controlled by ingested glucose as well as hormones and neural factors, therefore it is impossible to reproduce the physiological secretory pattern of insulin via exogenous insulin, even by multiple or continuous delivery by injection. Transplantation of beta cells has long been expected as the fundamental treatment to cure type 1 diabetics, and transplantation of the whole pancreas, both exocrine pancreas and islets, has been applied with success, resulting insulin independence. However, the exocrine pancreas, which releases amylase and trypsin to the digestive tract, is not indispensable for insulin replacement, so the interest in islet transplantation has increased enormously. In the past 20 years, the techniques for isolating large numbers of human islets have been advanced and more potent immunosuppressive agents have also been introduced, permitting newer attempts at islet transplantation. In 2000, insulin independence was first achieved in Canada using the Edmonton protocol. The success rates have increased gradually using this protocol, and 5 institutes in Japan have started to prepare human islet transplantation under the control of the Japan Pancreas and Islet Transplant Society. In 2004, insulin independence by islet transplantation was first achieved at Kyoto University Hospital and the number of islet transplantations has increased, though very slowly. By the end of 2005, approximately 100 patients were on the waiting list for islet transplantation in Japan. Many problems remain unsolved in islet transplantation to meet clinical practice: these are the shortage of insulin-producing cells, further progress in immunosuppressive agents that do not interfere with insulin secretion, strategies for protecting islets against hypoxia, and/or non-immunological damage such as mechanical damage soon after transplantation, and the destruction of islet cells by innate immunity. We investigated the role of neutrophil leucocytes infiltrating the liver using natural killer cell-knock-out mice and proposed a new strategy for protecting transplanted islets from non-specific damage, which occurs within one day after transplantation.

[Genetic screening for alpha-thalassemia deletional determinants by GapPCR method].

The majority of a-thalassemia results from the large deletions in a-globin gene cluster, including both or either one of alpha-globin genes (alpha1 and alpha2). Most common a-thalassemia-2 deletions (single gene deletions) are -alpha3.7 and -alpha4.2, and alpha-thalassemia-1 deletions (double gene deletions) are --SEA, --THAI, --FIL, --MED and -(alpha)20.5 Although it is not easy to diagnose these deletions because of the high GC content at this locus and the sequence homology among psi alpha2, psi alpha1, alpha2 and alpha1 genes, these alleles can now be diagnosed by a single tube multiplex gapPCR assay. We showed here two Saudi Arabian patients with a-thalassemia trait who could be determined their gene mutations according to the method of Chong SS et al. (2000). [Case 1: 21-year-old male] GapPCR assay revealed the amplification of only -alpha3.7, whereas PCR of both a-globin genes showed no amplifications. The results indicate case 1 is a homozygote of -alpha3.7(-alpha3.7/-alpha3.7). [Case 2: 31-year-old male] GapPCR assay revealed the amplification of only -alpha3.7, and PCR of both alpha globin genes showed normal amplification. DNA sequencing of the amplified a-globin genes revealed a point mutation in the poly A site of alpha2-globin gene (AATAAA-->AATAAG), which is known as alpha(T-Saudi). Thus, case 2 was confirmed to be a compound heterozygote of -alpha3.7 and alpha(T-Saudia) alpha(-alpha3.7 / alpha(T-Saudi) alpha). This gapPCR assay is a rapid, reliable screening test for common alpha-thalassemia deletions and seems to be useful for the diagnosis of thalassemic patients without an increase of Hb A2 and/or an abnormality of beta-globin gene.

Japanese standard reference material for JDS Lot 2 haemoglobin A1c. I: Comparison of Japan Diabetes Society-assigned values to those obtained by the Japanese and USA domestic standardization programmes and by the International Federation of Clinical Chemistry reference laboratories.

BACKGROUND: The Committee on Standardization of Laboratory Testing Related to Diabetes Mellitus of the Japan Diabetes Society (JDS) previously recommended use of the primary calibrator (JDS Lot 1) prepared by the former Committee for Standardization of Glycohemoglobin for standardizing the measurement of haemoglobin A1c (HbA1c). Owing to the depletion of vials of Lot 1 in March 2001, the present committee certified a new reference material, Lot 2, now distributed by the Health Care Technology Foundation (HECTEF). The standardization programme for HbA1c measurement in Japan is currently based on Lot 2, which has values assigned from within Lot 1; the Lot 1 values were consensus values based on assays by laboratories in the Japanese national quality control programme. In this study, for the purpose of international comparison and standardization, Lot 2 was assayed by the JDS reference laboratories, the National Glycoprotein Standardization Program (NGSP) in the USA, and by reference laboratories approved by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). METHOD: The HbA1c values of JDS Lot 2 were transferred from those assigned to Lot 1 using KO500, a high-resolution HPLC method, at three laboratories approved by the JDS committee. Subsequently, vials of JDS Lot 2 were shipped to and assayed by the NGSP in the USA and 10 IFCC reference laboratories. RESULT: The JDS-assigned HbA1c values (from Lot 1) are 4.04 for Level 1, 5.38 for Level 2, 7.32 for Level 3, 9.88 for Level 4, and 12.63 for Level 5, all expressed as a percentage of total haemoglobin. The values obtained by NGSP and the IFCC laboratories gave the following formulas: NGSP value(%)=JDS value(%)+0.3%; IFCC value(%)=1.068xJDS value(%)-1.741%. CONCLUSION: Although the values obtained by the IFCC laboratories are significantly lower than the values assigned to Lot 2 by the JDS, the relationship is linear. In addition, standardization of HbA1c based on JDS Lot 2 is currently at a satisfactory level in Japan. As a result, the reassignment of values for Lot 2 to agree with the IFCC values should be relatively easy and will be done after all relevant parties agree to the change.