Abnormal Excretion Research Articles

Nephroprotective Effect of Methanolic Extract of Solanum nigrum (Mako) and its Fractions on Cisplatin-induced Nephrotoxicity in Albino Rats

Background: Renal failure is an emerging problem worldwide and it is defined as the presence of renal parenchymal damage. Its manifestations are abnormal albumin excretion or decreased kidney function. Solanum nigrum L. (Mako), ‘Black Nightshade’, belongs to the family Solanaceae known in Unani medicine for its nephroprotective activity, especially in renal disease, as the berries are used to treat many renal ailments including renal failure. Aim: The aim of this study was to evaluate the nephroprotective activity of the berries of Mako on cisplatin-induced nephrotoxicity in rats. Methods: Wistar rats were divided into 7 groups of six animals in each. Group I served as the control, group II the toxicant group (cisplatin only), group III included the methanolic extract of Mako, group IV petrol soluble fraction of ME, group V chloroform soluble fraction of ME, group VI water soluble fraction of ME, and group VII the water soluble fraction of ME. All groups except for control group received a single dose of cisplatin 7 mg/kg injection on the 4th day, and test drugs were co-administered from day 1 to day 7. On the 8th day, samples were collected for the analysis of blood urea nitrogen (BUN), serum creatinine and for histopathological examination of renal tissues. Results: Results were analyzed using the one-way analysis of variance followed by Dunnett’s post hoc test. It was revealed that the ME of Mako and its fractions significantly reduced the rise in BUN and serum creatinine levels caused by cisplatin. The histopathological analysis also showed the protective nature of Mako in the nephrotoxic modules. Conclusion: Hence, the biochemical parameters and histopathological results affirmed the nephroprotective effect of the berries of the Mako against cisplatin-induced renal damage.

Modern Approaches to the Management of Patients with Hyperkaliemia

Hyperkalemia is the most common electrolyte imbalance in clinical practice. Hyperkalemia can be caused by an increased intake of potassium into the body, the shift of potassium out of cells or an abnormal renal potassium excretion. This condition is associated with a high risk of death from arrhythmias; therefore, even a slight deviation of the serum potassium level from the norm requires immediate correction. Modern approaches to the treatment of hyperkalemia include the elimination of predictors and the potassium-lowering drugs. Although inhibitors of the renin-angiotensin-aldosterone system are currently the best cardionephroprotective drugs, their administration can lead to hyperkalemia too, especially in heart failure, chronic kidney disease and diabetes mellitus. The article discusses in detail the physiology of potassium metabolism, possible predictors, prevention and treatment of hyperkalemia.

Association between Inguinal Hernia and Arterial Disease: A Preliminary Report.

Simple SummaryWhile the association between venous disease and inguinal hernia has been well demonstrated, there is less evidence concerning the association between arterial diseases (AD), such as carotid stenosis, peripheral artery disease and abdominal aortic aneurysms, and inguinal hernia. We surprisingly found that the prevalence of AD is large, being higher than 40% in our study cohort. Moreover, patients with AD as compared to those without AD are characterized by additional other comorbidities such as greater albuminuria, higher frequency of a smoking habit and older age. Hence, we provided a characterization of patients with inguinal hernia with respect to concomitant presence of AD. Background: Inguinal hernia (IH) is a major problem in general surgery and its prevalence is increasing. The presence of hernias has been associated with a wide spectrum of venous diseases, with the involvement of imbalances in collagen and extracellular matrix deposition and metalloproteinases dysfunction. We aimed to evaluate whether the association between IH and vascular diseases is also present with respect to arterial diseases. Methods: We designed a cross-sectional observational study enrolling consecutive patients undergoing surgical repair of IH. Arterial diseases (AD) considered were carotid stenosis, peripheral artery disease and abdominal aortic aneurysms. Results: Study population consisted of 70 patients. Mean age was 63.2 ± 4.7 years. Prevalence of AD was 42.9% in the whole cohort. AD patients were older (p = 0.015), and more frequently had hypertension (p = 0.001) and active smoking habits (p = 0.001) than the no-AD group. Albumin-to-creatinine ratio (ACR) was higher in AD than in no-AD patients (p < 0.001). At multivariable analysis, increased ACR (odds ratio, OR: 1.14, p < 0.001), old age (OR: 1.25, p = 0.001) and a smoking habit (OR: 3.20, p = 0.001) were significant correlates for the presence of AD. Conclusions: Prevalence of AD in patients with IH is non-negligible. Old age, a smoking habit and an abnormal excretion of urine albumin are associated with the presence of AD in these patients. Future studies are needed to gain more insights into the pathogenic mechanisms underlying this association, exploring also the specific role of metalloproteinases.

Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) detection as a rapid and convenient screening test for cystinuria

BackgroundCystinuria is an inborn congenital disorder characterised by a defective cystine metabolism resulting in the formation of cystine stones.The Brand’s test, used for diagnosis, requires dangerous substances, so has been replaced with high-performance liquid chromatography with fluorimetric detection (HPLC-FL). However, this technique requires the use of complex equipment.Infrared spectroscopy, universally used for stone analysis, recently was employed to detect insoluble cystine in urine. The aim of this study is to evaluate Infrared Spectroscopy combined with chemometric analysis as screening method to identify those patients requiring confirmation by HPLC-FL analysis. MethodsWe examined 24 h urine specimens from 57 patients. The quantitative analysis was performed by HPLC-FL. The infrared spectroscopic urine sediment analysis was performed with an ATR accessory (ATR-FTIR). Urine is centrifuged, the supernatant is discarded, and the sediment is dried on to the ATR prism surface.Statistical analysis was performed using a custom-made software developed in MATLAB environment. ResultsThe HPLC-FL determination showed a normal excretion of cystine in 49 samples and an abnormal excretion in the remaining 8 samples.The ATR-FTIR analysis combined with a statistical approach gives a sensitivity of 1.0 and a specificity of 0.82 were obtained. ConclusionsThe introduction of the ATR-FTIR technique in our clinical laboratory setting may reduce time and cost analysis for diagnosis of cystinuria.

Anesthetic management for mastectomy and total hysterectomy in a 49-year-old woman with type 1 sialidosis: a case report

BackgroundSialidosis is an autosomal recessive glycoprotein storage disorder, caused by neuraminidase deficiency which leads to abnormal intracellular accumulation and urinary excretion of sialylated oligosaccharides, resulting in various morphological and functional disorders. Only a few reports have described the anesthetic managements of patients with sialidosis.Case presentationA 49-year-old woman with type 1 sialidosis suffered from all limb contractures, an ocular cherry-red spot, and myoclonic seizures of the limbs. She had been cognitively normal. She was separately scheduled for mastectomy under total intravenous anesthesia and total hysterectomy under combined general and epidural anesthesia uneventfully.ConclusionsOur patient with type 1 sialidosis received both general and epidural anesthesia uneventfully. Anesthesiologists should carefully assess patients with sialidosis and give careful consideration to individually tailored anesthetic managements.

The effect of cranioskeletal trauma complicated by blood loss on the functional state of the liver in the early period of traumatic disease in rats with different resistance to hypoxia and their correction

Introduction. Traumatic events are considered to be one of the current problems in modern urban society. Beyond being the immediate cause of the death of the injured, the development of multiple organ failure syndrome is a perilous complication of severe multiple and combined trauma. An experimental cranioskeletal trauma is known to be accompanied by internal organ dysfunction. Nonetheless, the development of organ dysfunction in terms of hypoxia resistance in the presence of cranioskeletal trauma remains insufficiently studied.The objective of research: to establish the dynamics of the functional state of the liver in the presence of cranioskeletal trauma, complicated by blood loss in rats with different hypoxia resistance in the early period of traumatic disease and evaluate the efficacy of Thiocetam in the correction of identified abnormalities.Material and methods: The experimental studies were conducted on 196 white non-linear male rats weighing 180-200g. Initially, an individual resistance to hypoxia of the rats was estimated, and the animals demonstrated high- and low- resistance value to hypoxia (HR and LR, respectively) were selected for the further study. The HR- and LR-rats were separately divided into 4 groups: control and three experimental once. In the first experimental groups the HR-and LR-animals were induced the cranioskeletal trauma under thiopental sodium anesthesia (40 mg·kg-1); the animals of the second experimental groups were inflicted the acute blood loss of 20-22% of circulating blood volume; in the third experimental groups, the HR and LR animals were subjected to an acute blood loss following the cranioskeletal trauma and administered the intraperitoneal injection of the Thiocetam at a dose of 250 mg·кg-1 of body weight once a day for correction. The bile excretion function of the liver was analyzed in the controls and experimental groups of animals 1, 3 and 7 days after trauma.The results and discussion. The conducted studies indicated the higher rate of bile excretion in the control group of LR-rats compared to the control of HR-rats. The rate of bile excretion was occurred decreased in both HR- and LR-rats under the influence of the cranioskeletal trauma. However, the degree of decrease in the studied parameter of the LR-rats was considerably greater than the corresponding value of the HR-rats. An additional blood loss model resulted in more marked abnormalities, particularly in the experimental group of LR-rats. The 7-day administration of Thiocetam led to the significant decrease in the abnormal bile excretion rate in the experimental groups of HR- and LR-rats compared to the animals without corrective medication. The analysis of deviation degree of the studied parameter in the animals administered Thiocetam in relation to the animals without correction evidenced better efficacy of the medication in the experimental group consisted of LR-rats compared to the group of HR-rats, especially under the conditions of additional acute blood loss model. Consequently, Thiocetam is able to compensate less-developed defense mechanisms of LR-rats compared to the HR-rats, which should be taken into consideration in real-life clinical practice settings in the comprehensive treatment of the injured with cranioskeletal trauma complicated by blood loss considering the possibility to determine their resistance to hypoxia.Conclusions. The rate of bile excretion as a basic indicator of the functional state of the liver in the intact LR-rats is found to be substantially higher than in HR-rats. The degree of decrease in the rate of bile excretion is significantly greater in LR-rats after 7 days of the post-traumatic period under the influence of cranioskeletal trauma complicated by acute blood loss. The administration of Thiocetam is accompanied by a marked positive effect on the rate of bile excretion in the HR- and LR-rats, but the degree of increase in the studied parameter is considerably greater in the experimental group of LR-rats.

Renal Tubular Acidosis and Management Strategies: A Narrative Review

Renal tubular acidosis (RTA) occurs when the kidneys are unable to maintain normal acid−base homeostasis because of tubular defects in acid excretion or bicarbonate ion reabsorption. Using illustrative clinical cases, this review describes the main types of RTA observed in clinical practice and provides an overview of their diagnosis and treatment. The three major forms of RTA are distal RTA (type 1; characterized by impaired acid excretion), proximal RTA (type 2; caused by defects in reabsorption of filtered bicarbonate), and hyperkalemic RTA (type 4; caused by abnormal excretion of acid and potassium in the collecting duct). Type 3 RTA is a rare form of the disease with features of both distal and proximal RTA. Accurate diagnosis of RTA plays an important role in optimal patient management. The diagnosis of distal versus proximal RTA involves assessment of urinary acid and bicarbonate secretion, while in hyperkalemic RTA, selective aldosterone deficiency or resistance to its effects is confirmed after exclusion of other causes of hyperkalemia. Treatment options include alkali therapy in patients with distal or proximal RTA and lowering of serum potassium concentrations through dietary modification and potential new pharmacotherapies in patients with hyperkalemic RTA including newer potassium binders.

Statistical Optimization of Urinary Organic Acids Analysis by a Multi-Factorial Design of Experiment

The analysis of urinary organic acids is useful for patients suspected to have inborn errors of metabolism known as organic acidurias. These diseases cause an accumulation of organic acids in body fluids and their abnormal excretion in urines. By means of chemometrics tools, such as principal component analysis and multiple linear regression, it was concluded that the conditions used in our laboratory are really the most suitable to achieve high yields of analytes.

Ouabain Protects Nephrogenesis in Rats Experiencing Intrauterine Growth Restriction and Partially Restores Renal Function in Adulthood.

Intrauterine growth restriction (IUGR) is, in general, accompanied by a reduction of the nephron number, which increases the risk of hypertension and renal dysfunction. Studies have revealed that ouabain can partially restore the number of nephrons during IUGR. However, there is limited information regarding the melioration of nephric structure and function. We used maternal malnutrition to induce an IUGR model in rats. Subsequently, we used a mini-pump to administer ouabain to IUGR rats during pregnancy. Male offspring were divided randomly into two groups. One group was fed a normal diet, whereas the other was fed an isocaloric 8% high-salt diet. Maternal malnutrition led to a reduction in the birth weight and number of nephrons in offspring. At the end of a 40-week follow-up period, offspring from the IUGR group had high blood pressure and abnormal excretion of urinary protein; these parameters were exacerbated in offspring fed a high-salt diet. However, ouabain administration during pregnancy could partially restore the number of nephrons in IUGR offspring, normalize blood pressure, and reduce urinary protein excretion, even when challenged with a high-salt diet. Pathology findings revealed that IUGR, particularly following feeding of a high-salt diet, damaged the ultrastructure of glomeruli, but these harmful effects were ameliorated in offspring treated with ouabain. Collectively, our data suggest that ouabain could rescue nephrogenesis in IUGR newborns and protect (at least in part) the structure and function of the kidney during adulthood even when encountering unfavorable environmental challenges in subsequent life.

The Efficacy of Puromycin and Adriamycin for Induction of Glomerular Failure in Larval Zebrafish Validated by an Assay of Glomerular Permeability Dynamics.

Defects in the glomerular filtration barrier (GFB) play a major role in the onset of human renal diseases. Highly ramified glomerular cells named podocytes are a critical component of the GFB. Injury to podocytes results in abnormal excretion of plasma proteins, which can lead to chronic kidney disease. The conserved paired nephron of larval zebrafish is an excellent model for assessing glomerular function and injury. The efficacy of two known podocyte toxins was tested to refine models of acute podocyte injury in larval zebrafish. The validated compound was then used to test a novel assay of the dynamics of abnormal protein excretion. Injected adriamycin was found to be unsuitable for induction of glomerular injury due to off-target cardiovascular toxicity. In contrast, puromycin treatment resulted in a loss of discriminative filtration, measured by excretion of 70 kDa dextran, and podocyte effacement confirmed by electron microscopy. The dynamics of dextran excretion during puromycin injury modeled the onset of glomerular damage within 24 hours postinjection. These data validate puromycin for induction of acute podocyte injury in zebrafish larvae and describe a semihigh-throughput assay for quantifying the dynamics of abnormal protein excretion.

The urinary cortisol metabolome in patients with adrenal insufficiency: Dual-release hydrocortisone is less deleterious than conventional hydrocortisone therapy

Introduction Oral once-daily dual-release hydrocortisone (DR-HC) replacement therapy provides a more physiological cortisol profile than conventional thrice-daily (TID) therapy and has demonstrated improved metabolic profile among patients with adrenal insufficiency (AI). The mechanisms by which this metabolic improvement occurs may be due to less total exposure, changed cortisol time exposure profile, but also modified metabolism of cortisol. Objective To study cortisol metabolism during DR-HC and TID. Methods Patients with primary AI received DR-HC or an equal total daily dose of TID in a crossover multi-center study. Cortisol metabolites were measured by gas chromatography/mass spectrometry on 24 h urinary collections after both treatments and in controls. Results Fifty patients (22 females, mean age 47 years) and 124 healthy controls (73 females, mean age 48 years) were included in the study. Total cortisol metabolites were significantly decreased during DR-HC [median: 6380 μg/24 h] compared to TID [8825 μg/24 h]; P Conclusion The urinary cortisol metabolome may be a more sensitive marker of “optimal cortisol replacement”. Abnormal cortisol metabolites excretion and cortisol metabolism are observed in patients receiving TID. This abnormal profile improves with DR-HC. Especially, reduced 11βHSD1 activity during DR-HC may mediate some of the beneficial metabolic effects previously observed with this treatment.

The urinary cortisol metabolome in patients with adrenal insufficiency: dual-release hydrocortisone is less deleterious than conventional hydrocortisone therapy

Introduction Oral once-daily dual-release hydrocortisone (DR-HC) replacement therapy provides a more physiological cortisol profile than conventional thrice-daily (TID) therapy and has demonstrated improved metabolic profile among patients with adrenal insufficiency (AI). The mechanisms by which this metabolic improvement occurs may be due to less total exposure, changed cortisol time exposure profile, but also modified metabolism of cortisol. Objective To study cortisol metabolism during DR-HC and TID. Methods Patients with primary AI received DR-HC or an equal total daily dose of TID in a crossover multi-center study. Cortisol metabolites were measured by gas chromatography/mass spectrometry on 24 h urinary collections after both treatments and in controls. Results Fifty patients (22 females, mean age 47 years) and 124 healthy controls (73 females, mean age 48 years) were included in the study. Total cortisol metabolites were significantly decreased during DR-HC [median: 6380 μg/24 h] compared to TID [8825 μg/24 h]; P Conclusion The urinary cortisol metabolome may be a more sensitive marker of “optimal cortisol replacement”. Abnormal cortisol metabolites excretion and cortisol metabolism are observed in patients receiving TID. This abnormal profile improves with DR-HC. Especially, reduced 11βHSD1 activity during DR-HC may mediate some of the beneficial metabolic effects previously observed with this treatment.

Long-term intra-individual variability of albuminuria in type 2 diabetes mellitus: implications for categorization of albumin excretion rate

BackgroundDiabetic kidney disease (DKD) is the leading cause of end-stage renal disease in the Western world. Early and accurate identification of DKD offers the best chance of slowing the progression of kidney disease. An important method for evaluating risk of progressive DKD is abnormal albumin excretion rate (AER).Due to the high variability in AER, most guidelines recommend the use of more than or equal to two out of three AER measurements within a 3- to 6-month period to categorise AER. There are recognised limitations of using AER as a marker of DKD because one quarter of patients with type 2 diabetes may develop kidney disease without an increase in albuminuria and spontaneous regression of albuminuria occurs frequently. Nevertheless, it is important to investigate the long-term intra-individual variability of AER in participants with type 2 diabetes.MethodsConsecutive AER measurements (median 19 per subject) were performed in 497 participants with type 2 diabetes from 1999 to 2012 (mean follow-up 7.9 ± 3 years). Baseline clinical characteristics were collected to determine associations with AER variability. Participants were categorised as having normo-, micro- or macroalbuminuria according to their initial three AER measurements. Participants were then categorised into four patterns of AER trajectories: persistent, intermittent, progressing and regressing. Coefficients of variation were used to measure intra-individual AER variability.ResultsThe median coefficient of variation of AER was 53.3%, 76.0% and 67.0% for subjects with normo-, micro- or macroalbuminuria at baseline. The coefficient of variation of AER was 37.7%, 66% and 94.8% for subjects with persistent, intermittent and progressing normoalbuminuria; 43%, 70.6%, 86.1% and 82.3% for subjects with persistent, intermittent, progressing and regressing microalbuminuria; and 55.2%, 67% and 82.4% for subjects with persistent, intermittent and regressing macroalbuminuria, respectively.ConclusionHigh long-term variability of AER suggests that two out of three AER measurements may not always be adequate for the optimal categorisation and prediction of AER.

A Comprehensive review on Biomarkers for assessing Diabetic Kidney Disease

Diabetic nephropathy affects the body’s ability to grab the energy found in food . Futher it leads to major complications, where Diabetic nephropathy is seen as one of the major , and dangerous complication of this disorder. Effective and early diagnosis is of critically helpful .Diagnostic tests must be developed which must be more convenient and reliable than those currently used, would be highly beneficial. Urine is used as diagnostic medium for easy and non-invasive detection of biomarkers. This review briefly explains few of most relevant urinary biomarkers that are used to monitor the development and progression of Diabetes with its complications. The mortality and morbidity ratio are increasing in greater proportion, the consequences are expected to be at higher risk in near future. In particular, biomarkers of renal dysfunction such as transferrin, type IV collagen and N-acetyl-s-D-glucosamide are more sensitive than urinary albumin, in the detection of development nephropathy and risk regarding of cardiovascular disease. Inflammatory mediators including tumour necrosis factor-a, transforming growth factor-s, vascular endothelial growth factor and Monocyte chemo-attractant protein-1(MCP-1),Oxidative Stress Markers such as 8-oxo-7,8-dihydro-2'-deoxyguanosine and Immunoglobulin M, Podocyte, Cystatin C, Ceruloplasmin are useful biomarkers for diagnosis or monitoring of diabetic complications, particularly kidney disease. Abnormal excretion of 8-dihydro-2'-deoxyguanosine results in the indication of increased systemic level of DNA damage in non –insulin dependent diabetic patients. A comprehensive literature search has been carried out in PubMed, Google scholar and articles pertaining to urinary biomarkers assessing diabetic kidney disease were selected for review.

Assessment of the validity of protein - osmolality ratio in a randomly collected urine specimen, in the estimation of proteinuria in children

Background Proteinuria is an important marker of kidney disease. Protein-creatinine ratio and protein-osmolality ratio in a spot urine sample have been proposed as alternatives to 24-hour urinary protein excretion, to simplify sample collection and minimize errors due to incorrect sample collection. Objective Study was to compare validity of urine protein-osmolality ratio and urine protein-creatinine ratio in a random urine sample against 24-hour urinary protein excretion, to detect proteinuria in a paediatric population. Methods A cross sectional descriptive study was conducted by recruiting 85 children with kidney disease from medical wards and nephrology clinic at Lady Ridgeway Hospital, as well as, 56 healthy children aged 3-12 years. Twenty four-hour urine samples and spot urine samples were collected from each participant. Urine protein-osmolality ratio and urine protein-creatinine ratio in the spot urine sample were determined and compared with 24-hour urinary protein excretion. Data was analysed using SPSS statistical package. Standard descriptive methods (mean, median, standard deviation etc.) were used to describe the measured parameters. Receiver Operator Characteristic (ROC) curves were created using protein-osmolality ratio and protein-creatinine ratio as test variables, and 24-hour urinary protein excretion as the ‘gold standard’ variable. Optimum cut of values, maximizing sensitivity and specificity, for protein-osmolality ratio and protein-creatinine ratio was determined using ROC curves. Results The optimal value discriminating normal from abnormal protein excretion was determined to be a protein-osmolality ratio of 0.38 mg/L: mOsm/kgH 2 O (sensitivity 85.7%, specificity 100%) and a protein-creatinine ratio of 28 mg/mmol (sensitivity 100%, specificity 94%). The cutoff value for discriminating mild from nephrotic proteinuria was to be a protein-osmolality ratio of 2.00 mg/L: mOsm/kgH 2 O (sensitivity 91.5%, specificity 100%) and a protein-creatinine ratio of 186 mg/mmol (sensitivity 93%, specificity 98.5%). Conclusions Both urine protein-creatinine ratio and urine protein-osmolality ratio can be used to determine nephrotic proteinuria. Urine protein-creatinine ratio was more sensitive than urine protein-osmolality ratio in detecting mild proteinuria from normal proteinuria.

"Classical organic acidurias": diagnosis and pathogenesis.

Organic acidurias are inherited metabolic diseases due to the deficiency of an enzyme or a transport protein involved in one of the several cellular metabolic pathways devoted to the catabolism of amino acids, carbohydrates or lipids. These deficiencies result in abnormal accumulation of organic acids in the body and their abnormal excretion in urine. More than 65 organic acidurias have been described; the incidence varies, individually, from 1 out of 10,000 to >1 out of 1000,000 live births. Collectively, their incidence approximates 1 out of 3000 live births. Among these disorders, methyl malonic aciduria, propionic aciduria, maple syrup urine disease and isovaleric aciduria are sometimes referred to as classical organic acidurias. In this review, we focused on the basic GC-MS-based methodologies employed in the diagnosis of classical organic acidurias and provided updated reference values for the most common involved organic acids. We also attempted to provide the most recent updates on the pathogenetic bases of these diseases.

Enantioseparation of d,l-2-hydroxyglutaric acid by capillary electrophoresis with tandem mass spectrometry—Fast and efficient tool for d- and l-2-hydroxyglutaracidurias diagnosis

A novel capillary electrophoresis–tandem mass spectrometry method for the enantioseparation and identification of 2-hydroxyglutaric acid enantiomers without derivatization for clinical purposes was described. Vancomycin chloride was used as an efficient chiral selector for the discrimination of 2-hydroxyglutaric acid enantiomers by capillary electrophoresis employed complete capillary filling method. The obtained resolution was 2.05. Hyphenation of CE with tandem mass spectrometry allows a reliable identification of separated enantiomers as well as their quantification. The method was validated and applied for the separation, identification and determination of 2-hydroxyglutaric enantiomers in urine samples obtained from healthy patients and two urine samples obtained from child patients suffering from high urine excretion of 2-hydroxyglutaric acid. Abnormal excretion of d-hydroxyglutaric acid was found in both child urine samples (104.5±2.1 and 2200.0±12.6mmol/mol of creatinine, respectively). The limits of detection for d- and l-hydroxyglutaric acid were 31 and 38nmol/L, respectively.

P168 – 2679: Intrafamilial variability in Ehlers Danlos syndrome type VI

Objective The kyphoscoliotic type of the Ehlers-Danlos (EDSVIA) is a rare connective tissue disorder characterized by kyphoscoliosis, muscular hypotonia prominent in the neonatal period, joint hypermobility, easily bruisable skin, and occasionally rupture of the eye globe and vessels. Patients generally present with motor developmental delay. The clinical presentation among individuals affected with EDS VIA is variable, and genotype-phenotype correlation has not been established yet. Mutations in PLOD1 leading to deficiency of the enzyme lysyl hydroxylase 1 (LH1) cause underhydroxylation of collagen lysyl residues and, hence, abnormal formation of collagen cross-links. This results in an abnormal urinary excretion pattern of lysyl-pyridinolines (LP) and hydroxylysyl-pyridinolines (HP) cross-links. Therefore, the diagnosis of EDS VIA relies on the demonstration of a markedly increased ratio of total LP to HP (LP/HP), and may then be confirmed by molecular genetic analysis of PLOD1. Patients A 8.5-year-old girl, and her 11-year-old brother from a consanguineous family was evaluated because of developmental delay, scoliosis and laxity. Diagnosis at referral was Ullrich congenital muscular dystrophy. Skin lesions, joint hyper laxity, hyperelastic and fragile skin were additional clinical clues for EDSVIA. Conclusions We present a family with two affected siblings both showing increased urinary LP/HP ratio, harboring a homozygous large duplication (c.975+975_1755+? dup; p.Glu326_Lys585dup) between exon 10 and 16 of PLOD1, and showing broad intra-familial clinical variability especially in the degree of severity of kyphoscoliosis and gross-motor involvement.

P76 – 2450: Aspartylglucosaminuria: A rare cause of intellectual disability in children

Objective Aspartylglucosaminuria is a rare autosomal recessive lysosomal storage disorder characterized by the deficinecy of the enzyme aspartylglucosaminidase (AGA) leading early to a progressive intellectual disability. Aspartylglucosaminuria presents with progressive intellectual disability within the first years of life following a normal early infantile development. Aspartylglucosaminuria belongs to the group of disorders commonly referred to as the Finnish disease heritage, but has rarely been reported in Turkish patients. Case We report a 12 year-old boy who presented with psychomotor retardation and behavioral abnormalities. He was the third child of a couple who were first cousins. This 12 year old boy was the second child of a couple who were first cousins. He was born at term by spontaneous normal delivery. The parents were healthy. However, his mother had the history of 2 miscarriages in the first trimester. Though apparently normal at birth, their development milestones were slow. The boy showed mental retardation and mild motor delay. His speech was limited simple words (i.e. mum, dad). On physical examination, the boy appeared to have coarse facies, frontal bossing, prominent nose. The liver and the spleen were not palpable. Complementary test included: Karyotype, X-fragile: normal; blood and urine metabolic screening (lactate, pyruvate, ammonia, amino acids, organic acid, mucopolysacharides) normal; Urinary oligosaccharide analysis detected an abnormal excretion of aspartylglucosamine. Brain MRI showed J-shaped sella turcica, mildly widening of the lateral ventricles with thinning of the corpus callosum and enlarged perivascular spaces in the bilateral parietal region. Assay for AGA enzyme in fibroblasts reveal a very low activity, establishing a definitive diagnosis of AGU. Conclusions Clinical diagnosis of AGU is difficult; the initial presentation may be subtle and signs may be variable, resulting in frequent delays in diagnosis. Screening for aspartylglucosaminuria should be done in all patients with unexplained psychomotor retardation.

Effectiveness of GH isoform differential immunoassay for detecting rhGH doping on application of various growth factors

The analytical method for detecting growth hormone (GH) doping, the so-called GH isoform differential immunoassay, is currently approved by the World Anti-Doping Agency (WADA). Anti-doping laboratories often face challenges by athletes' lawyers and need to have various types of scientific evidence against the claim that the adverse analytical finding (AAF) result was caused by excess ectopic or abnormal excretion. In this work, a population study of Japanese athletes (255 male and 256 female) and administration studies of recombinant human GH (rhGH) in Japanese females were conducted to confirm the applicability of GH isoform differential immunoassay. The present paper describes the effectiveness of the GH isoform differential immunoassay under abnormal excretion of endogenous GH as determined by administration studies of GH releasing hormone (GHRH(1-44)) and insulin-like growth factor-1 (IGF-1). No false positive findings were found in Japanese athletes. The GH isoform differential immunoassays could detect application of rhGH for approximately 12-24 h. The administration of GHRH(1-44) and IGF-1 as well as ghrelin receptor agonists did not affect the isoform ratio (no false positives). We conclude that the GH isoform differential immunoassay is a highly specific method for detecting rhGH doping. Subject-based profiling (i.e. athlete biological passport) very likely will represent a highly sensitive approach for detecting rhGH doping.