The First Thai Case of Nondeletional HbH Disease Caused by Compound Heterozygosity for α-Thalassemia-1 Chiang Rai (--CR) Type Deletion with Hb Constant Spring.

Hemoglobin (Hb) H disease presents a wide range of clinical phenotypes, from asymptomatic to severe forms, depending on significant genetic heterogeneity. This is the first report of clinical and hematological features of the nondeletional HbH disease caused by --CR/αCSα. A baby was born to a father and a mother with --CR and αCSα carriers, respectively. She had severe symptomatic hypochromic microcytic anemia at 2 months of age with Hb 7.8 g/dL, packed cell volume (PCV) 0.27 L/L, mean corpuscular volume (MCV) 64.3 fL, and mean corpuscular Hb (MCH) 18.3 pg. The Hb analysis using capillary electrophoresis (CE) showed Hb Bart's, HbH, and Hb CS peaks at 17.1%, 2.2%, and 1.6%, respectively. A better understanding of a patient's clinical and hematological features with --CR/αCSα is useful for hemoglobinopathy counseling for the national thalassemia controlling program.

Impact of sediment mobilization on trace elements release in Galician Rías (NW Iberian Peninsula): insights into aquaculture.

In the latest years, the concentration levels of certain metals and metalloids in the sediments of the Galician Rías have shown an increasing trend (e.g., As, Zn, Cu, Pb, Hg). These areas are also characterized by their richness in nutrients and their great aquaculture or mariculture activity, with the presence of more than 3500 mussel rafts in the Rías Baixas. The inner areas of the Galician Rías are subjected to activities that resuspend the sediment such as high levels of maritime traffic and dredging or cleaning operations. It is likely that a transfer of these elements to the water column happens during the resuspension of sediments caused by natural events or anthropogenic activities. In this study, selected samples of surface sediments of the Ría de Pontevedra (NW Spain) were subjected to a procedure of aerobic oxidation to determine the concentration of some elements (Fe, Mn, Cu, Cr, Pb, Hg, and Zn) released from the sediment to the aqueous phase. The experiment was carried out within 5 days. Measurements of pH and total concentration were taken both in water and sediment samples. Furthermore, speciation of trace elements was carried out in the sediment samples. Trace element concentrations were lower in the sediments during aerobic oxidation, being released to the aqueous phase. From an environmental point of view, Cu was the only trace element released in quantities that may be toxic for the organisms in the area. This problem of sediment oxidation related to dredging activities or natural storm conditions should be considered in environmental impact studies and transferred to stakeholders.

Fate of microbial contamination in a South European Coastal Lagoon (Ria Formosa) under the influence of treated effluents dispersal.

AIM: Assessment of the fate of microbial contamination driven from treated wastewater disposal at a highly productive zone on a South European coastal lagoon (Ria Formosa). METHODS AND RESULTS: Microbial indicators of contamination (Total coliforms, Escherichia coli, and Enterococci) were evaluated monthly during September 2018-September 2020 at three study areas (Faro, Olhão, and Tavira) under different wastewater discharge flows and hydrodynamic conditions. Additional data on E. coli monitoring in bivalves, available from the national institution responsible for their surveillance was also considered. The maximum microbial contamination was found at Faro, the highest-load and less-flushed study area, contrasting the lowest contamination at Olhão, a lower-load and strongly flushed area. The wastewater impact decreased along the spatial dispersal gradients and during high water, particularly at Faro and Tavira study areas, due to a considerable dilution effect. Microbial contamination at Olhão increased during the summer, while at the other study areas seasonal evidence was not clear. Data also indicate that E. coli in bivalves from bivalve production zones next to the three study areas reflected the differentiated impact of the wastewater treatment plants effluents on the water quality of those areas. CONCLUSIONS: Effluent loads together with local hydrodynamics, water temperature, solar radiation, precipitation, and land runoff as well as seabirds populations and environmentally adapted faecal or renaturelized bacterial communities, contributed to microbial contamination of the study areas.

Geochemical and mineralogical characterization of Ria de Aveiro (Portugal) saltpan sediments for pelotherapy application.

The present study aims to characterize sediments textural, geochemical and mineralogical composition used in a SPA for pelotherapy applications. Six samples were collected in two areas of an open-air saline SPA, a former saltpan in Ria de Aveiro (Portugal). Sampling areas are predominantly composed by recent alluvium that had been affected by chemical industrial effluents for over 50 years. Samples < 2000, 63, and 2 µm fractions were analyzed by X-ray Diffraction (XRD) and X-ray Fluorescence (XRF), for identification of mineral phases and chemical composition. SEM analysis was used for individual particles morphological characterization and chemical semi-quantification. Texture, mineral phases and chemical composition showed the influence of SPA pond beneficiation works. The two SPA areas revealed distinct mineral phases, one with higher quartz content, and the other with higher halite content, consistent with their environmental conditions. Illite constituted the most abundant mineral phase of the clay fraction. Preliminary results suggested concentrations of potential toxic elements (e.g., As, Cd) above reference values, representing a risk to the ecosystem and humans.

Use of the 95-degree angled blade plate with biological and mechanical augmentation to treat proximal femur non-unions: a case series.

BACKGROUND: Intertrochanteric and subtrochanteric non-union are rare but challenging complications. In the present study, we investigate the use of a 95° blade, in association with biological and mechanical augmentation, in the management of intertrochanteric and subtrochanteric femoral non-unions. METHODS: Between October 2015 and February 2021, a retrospective cohort study was conducted at our institution to investigate the use of a 95° blade, in association with biological and mechanical augmentation, in the management of intertrochanteric and subtrochanteric femoral non-unions, following the mechanical failure of the first device. All the patients underwent a clinical and radiographic follow-up at 6 weeks, 3, 6, 9, 12 and 18 months; at each follow-up, a plain radiograph of the femur was performed and patients were assessed using Harris Hip Score (HHS) and the Short Form-12 (SF-12) questionnaire. RESULTS: From October 2015 and February 2021, 40 proximal femur non-unions were managed at our Institution. Fifteen patients out of forty (37.5%) met the inclusion criteria. The main data of the study are summarized in Table 1; patients' mean was 57 years old (range 19-83); 10 males and 5 females were included in the study. All the patients completely healed clinically and radiologically at an average of 6.1 months (range 4-13). All these patients returned to their pre-injury mobility status. During an average follow-up period of 25 months (range 8-60), the observed complications included wound dehiscence, which was treated with a superficial surgical debridement, a below-the-knee deep vein thrombosis, and a blade plate failure 3 months after the first revision surgery. CONCLUSIONS: This study shows the treatment of inter-and sub-trochanteric non-unions with a 95° blade plate, medial strut allograft, and bone autograft obtained with RIA system, together with a varus malalignment correction, leads to a high percentage of bone healing, with a low incidence of complications and good clinical outcome.

Management of bone defects using the Masquelet technique of induced membrane.

INTRODUCTION: Extensive bone defects are mostly caused by trauma, bone infection or bone tumours. The treatment is one of the most challenging issues of musculoskeletal surgery. Nowadays we have several techniques to deal with this problem. One of them is a two staged procedure described by Alain Masquelet, based on the principle of a foreign body-induced membrane with an autologous bone graft. The membrane, induced by a cement spacer, closes the defect and prevents resorption of the bone graft. In this paper we share our experience with this method. METHODS: We retrospectively analysed 10 patients who had been treated at our department between years 2012 and 2020. In addition to basic epidemiological data, we also collected data concerning the mechanism of injury, size of the defect, presence of infection and the infectious agent, type of spacer used and its retention, type of bone stabilization, time to full load, and time of full consolidation. RESULTS: We evaluated 10 patients with bone defects (5 of traumatic and 5 of infectious origin) operated at our department. Optimal healing free of complications was achieved in 50% of patients, and together with those where the 2nd phase had to be repeated, the success rate was 70%. Observed complications included two refractures, two cases of infection and one case of prolonged healing. CONCLUSIONS: Bone defect management using the modified Masquelet technique has an important place in reconstructive surgery. If the indication criteria and correct procedures are observed, combined with motivated and cooperating patients, the results are favourable, particularly in the group of non-infectious defects. Compared to other options of bone defect management, this technique is less demanding both for the patient and the surgeon.

Outcomes and complications of the reamer irrigator aspirator versus traditional iliac crest bone graft harvesting: a systematic review and meta-analysis.

BACKGROUND: The reamer irrigator aspirator (RIA) is a relatively recent device that is placed in the medullary canal of long bones to harvest a large volume of bone marrow, which is collected in a filtered canister. This study compares outcomes and complications of the RIA versus a traditional iliac crest bone graft (ICBG) for the treatment of bone defects. METHODS: This meta-analysis was conducted according to the PRISMA guidelines. The Embase, Google Scholar, PubMed, and Scopus databases were accessed in June 2021. All clinical trials comparing the RIA and ICBG with a minimum of 6 months follow-up were included. RESULTS: Data from 4819 patients were collected. The RIA group demonstrated lower site pain (P < 0.0001), fewer infections (P = 0.001), and a lower rate of adverse events (P < 0.0001). The ICBG group demonstrated a greater rate of bone union (P < 0.0001). There was no difference between groups in VAS (P = 0.09) and mean time to union (P = 0.06). CONCLUSION: The current evidence supports the use of the RIA, given its low morbidity and short learning curve.

High Nitrogen and Phosphorous Acquisition by Belowground Parts of Caulerpa prolifera (Chlorophyta) Contribute to the Species' Rapid Spread in Ria Formosa Lagoon, Southern Portugal.

Despite worldwide proliferation of the genus Caulerpa and subsequent effects on benthic communities, little is known about the nutritional physiology of the Caulerpales. Here, we investigated the uptake rates of ammonium, nitrate, amino acids, and phosphate through the fronds and rhizoids + stolon, the internal translocation of nitrogen, and developed a nitrogen budget for the rapidly spreading Caulerpa prolifera in Ria Formosa lagoon, southern Portugal. Caulerpa prolifera acquired nutrients by both aboveground and belowground parts at similar rates, except nitrate, for which fronds showed 2-fold higher uptake rates. Ammonium was the preferential nitrogen source (81% of the total nitrogen acquisition), and amino acids, which accounted for a significant fraction of total N acquisition (19%), were taken up at faster rates than nitrate. Basipetal translocation of 15 N incorporated as ammonium was nearly 3-fold higher than acropetal translocation, whereas 15 N translocation as nitrate and amino acids was smaller but equal in either direction. The estimated total nitrogen acquisition by C. prolifera was 689 µmol · m-2 · h-1 , whereas the total nitrogen requirement for growth was 672 µmol · m-2  · h-1 . The uptake of ammonium and amino acids by belowground parts accounted for the larger fraction of the total nitrogen acquisition of C. prolifera and is sufficient to satisfy the species nitrogen requirements for growth. This may be one reason explaining the fast spreading of the seaweed in the bare sediments of Ria Formosa where it does not have any macrophyte competitors and the concentration of nutrients is high.

Calcitonin levels by ECLIA correlate well with RIA values in higher range but are affected by sex, TgAb, and renal function in lower range.

Calcitonin (CT) is a marker for both initial diagnosis and monitoring of patients with residual or recurrent medullary thyroid carcinoma (MTC). In Japan, serum CT had been measured by radioimmunoassay (RIA) until recently. Electrochemiluminescence immunoassay (ECLIA) became commercially available in 2014, and this technique is now the only method used to examine CT concentration. The purposes of this study were to investigate the correlations between the CT concentration measured with ECLIA (ECLIA-CT) and RIA (RIA-CT) and to explore the clinical characteristics of patients with elevated ECLIA-CT. CT concentrations of 348 sera samples from 334 patients with various thyroid disorders including nine MTC were measured using both assays. The correlation analysis revealed an excellent correlation between ECLIA-CT and RIA-CT among the cases with CT level >150 pg/mL by both assays (rs = 0.991, p < 0.001). However, 63% of all samples exhibited undetectable ECLIA-CT, while their RIA-CTs were measured between 15 and 152 pg/mL. The ECLIA-CTs in all patients who underwent total thyroidectomy for non-MTC showed low concentrations. High ECLIA-CT was observed in patients with MTC or pancreas neuroendocrine tumor. ECLIA-CT was also increased in 14 other male patients with non-MTC, including four with renal failure. Multivariate logistic regression analysis showed that male sex, negative TgAb, and lower estimated glomerular filtration rate were independent factors to predict detectable ECLIA-CT (≥0.500 pg/mL). These results indicate that ECLIA-CT correlates well with RIA-CT in higher range and is affected by sex, TgAb, and renal function.

How do organisations implement research impact assessment (RIA) principles and good practice? A narrative review and exploratory study of four international research funding and administrative organisations.

BACKGROUND: Public research funding agencies and research organisations are increasingly accountable for the wider impacts of the research they support. While research impact assessment (RIA) frameworks and tools exist, little is known and shared of how these organisations implement RIA activities in practice. METHODS: We conducted a review of academic literature to search for research organisations' published experiences of RIAs. We followed this with semi-structured interviews from a convenience sample (n = 7) of representatives of four research organisations deploying strategies to support and assess research impact. RESULTS: We found only five studies reporting empirical evidence on how research organisations put RIA principles into practice. From our interviews, we observed a disconnect between published RIA frameworks and tools, and the realities of organisational practices, which tended not to be reported. We observed varying maturity and readiness with respect to organisations' structural set ups for conducting RIAs, particularly relating to leadership, skills for evaluation and automating RIA data collection. Key processes for RIA included efforts to engage researcher communities to articulate and plan for impact, using a diversity of methods, frameworks and indicators, and supporting a learning approach. We observed outcomes of RIAs as having supported a dialogue to orient research to impact, underpinned shared learning from analyses of research, and provided evidence of the value of research in different domains and to different audiences. CONCLUSIONS: Putting RIA principles and frameworks into practice is still in early stages for research organisations. We recommend that organisations (1) get set up by considering upfront the resources, time and leadership required to embed impact strategies throughout the organisation and wider research 'ecosystem', and develop methodical approaches to assessing impact; (2) work together by engaging researcher communities and wider stakeholders as a core part of impact pathway planning and subsequent assessment; and (3) recognise the benefits that RIA can bring about as a means to improve mutual understanding of the research process between different actors with an interest in research.

Development of in-house RIA kit for progesterone in cow's skim milk.

A radioimmunoassay (RIA) method for progesterone has been developed. It has been validated for the determination of progesterone in cow skim milk. The main reagents used in the development work were prepared and characterized in-house. The assay uses a radioiodinated tracer purified by gel filtration on Sephadex G-25, standards prepared in skim milk and coated tubes with specific antibodies as the solid phase (separation system). The radiochemical purity of the tracer was greater than 95%, the maximum binding using solid phase reached 43% and the nonspecific binding didn't exceed 5%. Series of progesterone standards using milk matrix with concentration ranging from 0 to 40ng/mL were prepared. Detection limit of the assay was 0.13ng/mL and the precision evaluation gives an intra and inter-assay coefficient variations between 4.94% and 12.66%. The recovery obtained with skim milk samples was 84% to 119% and the parallelism test indicated good linearity (R2 > 0.99). The clinical tests give high correlation coefficient (r = 0.998) between progesterone concentrations of cow's skim milk assayed using developed progesterone RIA kit and commercial progesterone RIA kit.

Bone Grafting in Total Wrist Arthrodesis With Large Bone Defects Using the Reamer-Irrigator-Aspirator: A Case Study of 2 Patients.

Total wrist arthrodesis, fusion of the carpus to the radius, is a well-established and commonly performed procedure for the management of pancarpal degenerative arthritis, trauma, and other severe diseases of the wrist. A well-known complication of this is radiocarpal nonunion, with a rate of 4.4%. Commonly, bone grafting is done at the time of the surgery using local bone, iliac crest bone graft, or cancellous allograft bone. The Synthes Reamer-Irrigator-Aspirator (RIA) is a flexible reamer originally designed as a single-pass intramedullary reamer that can be used in reaming of long bone fractures prior to insertion of an intramedullary nail. Recently, its use has been expanding to the harvesting of bone graft. We present a case study of 2 patients who had total wrist arthrodesis after severe trauma to the wrist resulting in severe bone loss, in whom we used an RIA to obtain bone graft, and both patients went on to union. The first patient is a 17-year-old male who sustained a blast injury from a firework resulting in a mangling injury to the wrist with soft tissue and severe bone loss of his carpus. After being treated with multiple debridements with an external fixator, he was definitively treated with a wrist fusion with RIA bone graft. He went on to osseous union by his 4-month follow-up. Our second patient is a 53-year-old woman who sustained a gunshot wound to the wrist resulting in massive bone loss of the distal radius. After being initially treated with multiple debridements and external fixation, she was treated with a wrist fusion with RIA bone graft and went on to osseous fusion by her 3-month follow-up. Neither patient had complications from the RIA procedure.

Meta-analysis across Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium provides evidence for an association of serum vitamin D with pulmonary function.

The role that vitamin D plays in pulmonary function remains uncertain. Epidemiological studies reported mixed findings for serum 25-hydroxyvitamin D (25(OH)D)-pulmonary function association. We conducted the largest cross-sectional meta-analysis of the 25(OH)D-pulmonary function association to date, based on nine European ancestry (EA) cohorts (n 22 838) and five African ancestry (AA) cohorts (n 4290) in the Cohorts for Heart and Aging Research in Genomic Epidemiology Consortium. Data were analysed using linear models by cohort and ancestry. Effect modification by smoking status (current/former/never) was tested. Results were combined using fixed-effects meta-analysis. Mean serum 25(OH)D was 68 (sd 29) nmol/l for EA and 49 (sd 21) nmol/l for AA. For each 1 nmol/l higher 25(OH)D, forced expiratory volume in the 1st second (FEV1) was higher by 1·1 ml in EA (95 % CI 0·9, 1·3; P<0·0001) and 1·8 ml (95 % CI 1·1, 2·5; P<0·0001) in AA (P race difference=0·06), and forced vital capacity (FVC) was higher by 1·3 ml in EA (95 % CI 1·0, 1·6; P<0·0001) and 1·5 ml (95 % CI 0·8, 2·3; P=0·0001) in AA (P race difference=0·56). Among EA, the 25(OH)D-FVC association was stronger in smokers: per 1 nmol/l higher 25(OH)D, FVC was higher by 1·7 ml (95 % CI 1·1, 2·3) for current smokers and 1·7 ml (95 % CI 1·2, 2·1) for former smokers, compared with 0·8 ml (95 % CI 0·4, 1·2) for never smokers. In summary, the 25(OH)D associations with FEV1 and FVC were positive in both ancestries. In EA, a stronger association was observed for smokers compared with never smokers, which supports the importance of vitamin D in vulnerable populations.

Direct comparison of LC-MS/MS and RIA methods for the pharmacokinetics assessment of human insulin in preclinical development.

Insulin is an effective therapeutic for diabetes, and the level of insulin in vivo is directly related to the health of diabetic patients. Traditionally, the concentrations of insulin in vivo are determined by the radioimmunoassay (RIA) method. In this study, we developed an LC-MS/MS method for the quantification of human insulin in dog plasma and directly compared the RIA and LC-MS/MS methods. Our LC-MS/MS method exhibited superior accuracy, efficiency and cost-effective for the pharmacokinetic (PK) assessment of human insulin. The LC-MS/MS method can quantitate human insulin and canine insulin simultaneously without cross-reactivity, making the analysis more efficient. The LLOQ of our LC-MS/MS method was 38.5 pg/mL, which was necessary to fully describe the PK profiles of endogenous and exogenous insulin in vivo. The direct comparison of PK data obtained from the two methods demonstrated that LC-MS/MS could be an alternative to the RIA method and should be widely used for the quantification of insulin drugs, especially in preclinical studies.

Correlation of two radioimmunoassay systems for measuring plasma pregnancy-associated glycoproteins concentrations during early pregnancy and postpartum periods in water buffalo.

This is the first time that PAG determination using two different antisera raised against PAG molecules purified from both caprine (RIA-706) and bubaline placentas (RIA-860) is reported in water buffalo. Ninety-eight buffalo cows, belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) programme, were enrolled in this study. Blood samples were taken on days 0 (AI), 23, 25, 28, 30 and 45. Pregnancy was confirmed by ultrasonography on days 28 and 45. The blood of 20 buffaloes that had calved was tested every five days from the day of calving until day 50 postcalving. Differences in PAG concentrations were observed between pregnant and nonpregnant buffaloes starting from day 23 post AI using both RIA-706 and RIA-860 (p < 0.001). However, estimated mean concentrations of PAG measured by RIA-706 were higher than RIA-860 (p < 0.001) and Bland-Altman analysis showed biases ranged from 0.0 ng/ml at day 23 to 0.79 ng/ml at day 28 post AI. Moreover, RIA-706 showed greater sensitivity and accuracy both at 23 and 25 days of pregnancy. RIA-706 and RIA-860 decreased below 1 ng/ml from 40 and 30 days postpartum, respectively, suggesting that PAG are better recognized by the antisera raised against the caprine PAG in the postpartum period also. This is essential when using PAG as an appropriate marker of early pregnancy after postpartum for detecting new pregnancies. The results of this study show that the ability of RIA systems to recognize early PAG could be improved using antisera raised against PAG molecules isolated from caprine placenta.

Insulin-Sensitizer Effects of Fenugreek Seeds in Parallel with Changes in Plasma MCH Levels in Healthy Volunteers.

In developed, developing and low-income countries alike, type 2 diabetes mellitus (T2DM) is one of the most common chronic diseases, the severity of which is substantially a consequence of multiple organ complications that occur due to long-term progression of the disease before diagnosis and treatment. Despite enormous investment into the characterization of the disease, its long-term management remains problematic, with those afflicted enduring significant degradation in quality-of-life. Current research efforts into the etiology and pathogenesis of T2DM, are focused on defining aberrations in cellular physiology that result in development of insulin resistance and strategies for increasing insulin sensitivity, along with downstream effects on T2DM pathogenesis. Ongoing use of plant-derived naturally occurring materials to delay the onset of the disease or alleviate symptoms is viewed by clinicians as particularly desirable due to well-established efficacy and minimal toxicity of such preparations, along with generally lower per-patient costs, in comparison to many modern pharmaceuticals. A particularly attractive candidate in this respect, is fenugreek, a plant that has been used as a flavouring in human diet through recorded history. The present study assessed the insulin-sensitizing effect of fenugreek seeds in a cohort of human volunteers, and tested a hypothesis that melanin-concentrating hormone (MCH) acts as a critical determinant of this effect. A test of the hypothesis was undertaken using a hyperinsulinemic euglycemic glucose clamp approach to assess insulin sensitivity in response to oral administration of a fenugreek seed preparation to healthy subjects. Outcomes of these evaluations demonstrated significant improvement in glucose tolerance, especially in patients with impaired glucose responses. Outcome data further suggested that fenugreek seed intake-mediated improvement in insulin sensitivity correlated with reduction in MCH levels.

Prevalence of HIV Infection and Risk Factors Among Female Sex Workers in a Southeast Province of Vietnam.

Female sex workers (FSWs) are at heightened risk of HIV infection. This research aims to determine the prevalence of HIV and relevant risk factors and related behavior among FSWs in Ba Ria - Vung Tau, a southeast province of Vietnam. 420 FSWs were interviewed using a structured questionnaire and biological samples tested for HIV. 2.6 % were found to be HIV positive. HIV infection was significantly higher in FSWs who had low income (≤AUD 200 per month), have had anal sex, have had sex with injecting drug users, and had a low level of HIV/AIDS-related knowledge. Improved employment opportunities and income are important to reduce the pressure for young women to engage in sex work for income purposes, but in public health terms, existing HIV treatment, prevention and intervention programs needs better targeting and improvements to reduce the risk of HIV infection.

Accurate analytical method for human plasma glucagon levels using liquid chromatography-high resolution mass spectrometry: comparison with commercially available immunoassays.

Accurate quantification of plasma glucagon levels in humans is necessary for understanding the physiological and pathological importance of glucagon. Although several immunoassays for glucagon are available, they provide inconsistent glucagon values owing to cross-reactivity of the antibodies with peptides other than glucagon. To overcome this limitation, we developed a novel method to measure glucagon levels by a liquid chromatography (LC)-high-resolution mass spectrometry (HRMS) assay via parallel reaction monitoring (PRM) without immunoaffinity enrichment. Using stable isotope-labeled glucagon as an internal standard and 200 µL of plasma, the lower limit of quantification was 0.5 pM. This method was applied to measure plasma glucagon levels during the oral glucose tolerance test (OGTT) and meal tolerance test (MTT) in healthy volunteers, and its results were compared with those of sandwich enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). During the OGTT, this method showed significant suppression of plasma glucagon levels, and similar patterns were observed with sandwich ELISA and RIA. In contrast, during the MTT, plasma glucagon levels were slightly elevated according to the LC-MS/MS and sandwich ELISA results and were reduced according to RIA results. Our newly developed LC-MS/MS method overcomes a lack of specificity among currently available immunoassays for glucagon and may contribute to a better understanding of the importance of glucagon. Graphical abstract Flowchart for the extraction and quantification of glucagon in human plasma, and plasma glucagon responses in healthy volunteers quantified by the present LC-MS/MS, sandwich ELISA, and RIA during OGTT and MTT.

Preparation and development of solid phase radioimmunoassay system for determination of cortisol.

The goal of this article was oriented to prepare stable polystyrene coated tubes for direct radioimmunoassay (RIA) of human cortisol. Coating process was performed using rabbit polyclonal antisera specific for cortisol. The stability study showed that these tubes could be stored for up to one year at 4ºC without any appreciable reduction in their binding. Labeling of cortisol was carried out using chloramin-T method and the 125I-cortisol tracer was purified using HPLC column. Preparation of polyclonal antibodies (anti-cortisol) was carried out in host rabbit animals against cortisol-21-hemisuccinate:Bovine serum albumin conjugate. The statistical analysis revealed good correlations between the results from the present study and the commercial IZOTOP RIA kit. The local may be extremely helpful in diagnosis and proper management of adrenal cortex disorders.

Standardization of Growth Hormone and Insulin-like Growth Factor-I Measurements.

Measurement of the levels of growth hormone (GH) and its related factor insulin-like growth factor I (IGF-I) is essential for the diagnosis and treatment of GH deficiency (GHD) and conditions related to excess GH such as acromegaly and pituitary gigantism. Measurement of GH levels is also used as an indicator of hypothalamic-pituitary function. Because of the marked variability in GH measurements among kits, the Study Committee for GH and Its Related Factors of The Foundation for Growth Science, Japan standardized GH values measured with various commercially available GH assay kits in Japan. The committee also established IGF-I reference values for Japanese subjects of all ages, from childhood to adulthood. Hopefully, international harmonization of GH measurements will be achieved in the future.