First report of carglumic acid in a patient with citrullinemia type 1 (argininosuccinate synthetase deficiency).

WHAT IS KNOWN AND OBJECTIVE: Carglumic acid is a structural analogue of human N-acetylglutamate, which has become an alternative therapeutic option for hyperammonaemia in organic acidaemias such as isovaleric acidaemia, methylmalonic acidaemia and propionic acidaemia, and it has been suggested in other urea cycle disorders such as ornithine transcarbamylase deficiency and carbamoyl phosphate synthetase 1 deficiency. CASE DESCRIPTION: A male newborn was diagnosed with citrullinemia after serum amino acid analyses revealed markedly elevated citrulline concentration together with homozygous p.Gly390Arg mutation in ASS1 gene. The ammonia concentration decreased and blood gas analysis normalized after peritoneal dialysis was performed for three days. Also, sodium benzoate, L-arginine and parenteral nutrition with glucose and lipid therapy were initiated. Until 1 year of age, low adherence to sodium benzoate therapy due to unpleasant taste caused hyperammonaemic episodes and obligated us to initiate carglumic acid (100 mg/kg/day) therapy. During treatment with carglumic acid, the median ammonia level was 45.6 µmol/L. The patient's treatment was switched from carglumic acid to sodium phenylbutyrate when he was 4.5 years old. Currently, the patient is 6.5 years old and remains under follow-up with sodium phenylbutyrate, L-arginine and protein-restricted diet. Plasma ornithine level was found to be significantly lower during the carglumic acid treatment compared to other treatments (P=.039). Also, glutamic acid was found to be higher during the sodium benzoate treatment period compared to other treatment periods (P=.024). WHAT IS NEW AND CONCLUSION: To the best of our knowledge, this is the first report describing the long-term use of carglumic acid in a patient with argininosuccinate synthetase deficiency.

Evaluation of prehypertension and masked hypertension rate among clinically normotensive patients.

BACKGROUND: The present cross-sectional study was aimed to identify pre-hypertension and masked hypertension rate in clinically normotensive adults in relation to socio-demographic, clinical and laboratory parameters. METHODS: A total of 161 clinically normotensive adults with office blood pressure (OBP) <140/90 mmHg without medication were included in this single-center cross-sectional study. OBP, home BP (HBP) recordings and ambulatory BP monitoring (ABPM) were used to identify rates of true normotensives, true pre-hypertensives and masked hypertensives. Data on sociodemographic and clinical characteristics were collected in each subject and evaluated with respect to true normotensive vs. pre-hypertensive patients with masked hypertension or true pre-hypertensive. Target organ damage (TOD) was evaluated in masked hypertensives based on laboratory investigation. RESULTS: Masked hypertension was identified in 8.7% of clinically normotensives. Alcohol consumption was significantly more common in masked hypertension than in true pre-hypertension (28.6 vs. 0.0%, p = 0.020) with risk ratio of 2.7 (95% CI 1.7-4.4). Patients with true pre-hypertension and masked hypertension had significantly higher values for body mass index, waist circumference, systolic and diastolic OBP and HBP (p < 0.05 for each) compared to true normotensive subjects. ABPM revealed significantly higher values for day-time and night-time systolic and diastolic BP (p = 0.002 for night-time diastolic BP, p < 0.001 for others) in masked hypertension than true pre-hypertension. CONCLUSIONS: Given that the associations of pre-hypertension with TOD might be attributable to the high prevalence of insidious presentation of masked hypertension among pre-hypertensive individuals, ABPM seems helpful in early identification and management of masked hypertension in the pre-hypertensive population.

Could immune cells be associated with nephropathy in Fabry disease patients?

BACKGROUND: In Fabry Disease (FD), although the primary factor initiating kidney damage is glycosphingolipid accumulation, secondary conditions such as increased inflammation and fibrosis may cause this damage to progress. These processes may be induced by immune cells. Therefore, we aimed to investigate the peripheral lymphocyte subgroup analysis of the patients with FD and compare these results with healthy individuals. In addition, we performed T, B, NK, and plasma cell analyses in kidney biopsy materials and compared these kidney biopsy results with the biopsy results of patients whose kidney functions were impaired after 4 years of regular ERT. MATERIALS AND METHODS: 18 FD and 16 healthy individuals were included in the study. T-B lymphocyte and NK-cell populations were determined. We performed kidney biopsies (KBx) on 13 patients with FD prior to ERT. Of these, 4 patients had rebiopsy after 4 years of regular ERT. Immunohistochemical staining was performed to define immune cell infiltration. RESULTS: There was no statistically significant difference in terms of total, helper and cytotoxic T-lymphocyte and CD3-CD16+CD56+ natural killer (NK)-cell count (p = 0.20; p = 0.12; p = 0.76; p = 0.75, respectively).According to KBx findings prior to ERT, all patients had interstitial fibrosis (IF), podocyte vacuoles (PV), and podocyte inclusion (PI), CD3, CD4, CD8, CD16, and CD56 positivity at different levels. None of the patients had CD19, CD20, and CD138 positivity at the first biopsies. When we compared the first and the second KBx results of the two progressors, we also demonstrated that CD3+4+T-cells infiltration remained the same, whereas CD8+T cells, CD16+ and 56+NK-cells infiltration were significantly decreased. In contrast, CD20+B cells and CD138+plasma cell infiltration were significantly increased despite 4 years of ERT (15 fold and sixfold, respectively). The CD20+B and CD138+ plasma cells and IF were positively correlated with proteinuria. CONCLUSIONS: The progression of FD nephropathy and proteinuria is increased despite a long-term ERT. Immune cells, primarily B and plasma cells, might cause these unwanted consequences.

Combined liver-kidney transplantation and follow-up in primary hyperoxaluria treatment: report of three cases.

INTRODUCTION: Primary hyperoxaluria type-1 (PH1) is an autosomal recessive disorder caused by impaired activity of the hepatic peroxisomal alanine-glyoxilate aminotransferase, which leads to end-stage renal disease (ESRD) and requires combined liver-kidney transplantation (CLKT). Herein, we have reported 3 children diagnosed with PH1 who received CLKT. CASE 1: A 4.5-year-old boy with an elder brother diagnosed with PH1 was diagnosed during family screening when the sonography showed multiple calculi. Within 5 years he experienced flank pain, hematuria attacks, and anuric phases due to obstruction and received hemodialysis (HD) when ESRD appeared. CLKT was performed from his full-match sister at the age of 9.5. He is doing well at 5.5 years. CASE 2: A 7-year-old boy was admitted with polyuria, polydypsia, and stomach pain with renal stones on sonography. PD was instituted when serum creatinine and BUN levels were measured as high values. At the age of 10, CKLT was performed from his mother. His liver and renal function tests are well at 14 months after CKLT. CASE 3: A 2.5-year-old girl had attacks of dark urine without any pain; renal stones were imaged on sonography. She was diagnosed with PH1 and operated on several times due to obstruction. She received peritoneal dialysis and a cadaveric CLKT was performed when she was 9 years old. At the age of 16, she experienced chronic allograft nephropathy requiring HD and subsequent cadaveric donor renal transplantation at 1.5 years after initiation of HD. CONCLUSION: Herein, we have presented the favorable clinical outcomes of patients with CKLT to indicate the validity of this treatment choice for PH1.

Multiorgan involvement in a pediatric patient with hydatid disease.

Human Echinococcus infection still remains an important health problem in endemic regions. Herein, we report a 5-year-old boy with hydatid disease who has spleen, lung, kidney and liver involvement simultaneously. To our knowledge, there is no pediatric case with hydatid disease in the literature reporting simultaneous involvement of spleen, kidney, liver and lungs as in our case.

Haemostatic actions of the folkloric medicinal plant extract Ankaferd Blood Stopper.

Ankaferd Blood Stopper (ABS), a standardized mixture of five plants, has been used historically as a haemostatic agent but its mechanism of action remains unknown. This study investigated the in vitro effects of ABS on haemostatic parameters. When added to plasma or serum, ABS induced the very rapid formation of a protein network and erythrocyte aggregation. The levels of coagulation factors II, V, VII, VIII, IX, X, XI, and XIII were not affected by ABS. Plasma fibrinogen activity and antigen levels were decreased following the addition of ABS, in parallel with the prolonged thrombin time. Total protein, albumin, and globulin levels decreased after the addition of ABS. Our findings suggest that ABS stimulates the formation of an encapsulated protein network that provides focal points for erythrocyte aggregation. ABS has the therapeutic potential to be used for the management of haemorrhage and this agent should be investigated further in clinical trials.

Phytochemical and antioxidant analysis of eight Hypericum taxa from Central Italy.

Eight taxa of the Hypericum spp. growing in Central Italy (Appennino Umbro-Marchigiano) were analyzed by HPLC-DAD for constituents quantitation, for antioxidant and free radical scavenging activities. H. perforatum subsp. veronense was the richest in phenolic compounds and hyperforin was detected for the first time in H. hircinum subsp. majus. Significant values of antioxidant activity were found in the investigated Hypericum taxa.

Evaluation of topical Dexmedetomidine administration in postlaminectomy epidural fibrosis rat model.

Epidural fibrosis is a challenging topic in spinal surgery. Numerous clinical and experimental studies have been focused on this issue to clarify problems faced in spinal procedures for the patient as well as the surgeon and find out new methodologies. Dense cytokines and growth factors which are released from inflammatory cells have been suggested to play a major role in the inception and progression of fibrosis. One of the most investigated and important actor in epidural fibrosis is assumed to be the transforming growth factor-1ß (TGF-1ß) formation. Studies showed that Dexmedetomidine (DEX) downregulates TGF-ß pathway with its anti-inflammatory and antioxidant effects. From this point of view, for the first time in the literature we try to observe if there will be an effect of topical DEX administration over epidural fibrosis in a rat model. We hypothesized that DEX might have preventive effects on epidural fibrosis via anti-inflammatory and antioxidant effects. Twenty-four adult male Wistar albino rats were randomly assigned to three groups (Topical DEX, Spongostan, Laminectomy). A total laminectomy was performed at the L3-L5 level and then the ligamentum flavum and epidural fat tissue were cleared away from the surgical site. Histopathological assessment was performed postoperatively after 4 weeks. Our study revealed that topical DEX administration may have effects on reducing epidural fibrosis. Topical DEX administration may be helpful in preventing epidural fibrosis after laminectomy in rats through multiple anti-inflammatory and antioxidant mechanisms as well as through TGF -1ß pathway.

Note: 3D printed spheroid for uniform magnetic field generation.

This article is focused on a novel and practical production method for a uniform magnetic field generator. The method involves building of a surface coil template using a desktop 3D printer and winding of a conducting wire onto the structure using surface grooves as a guide. Groove pattern was based on the parametric spheroidal helical coil formula. The coil was driven by a current source and the magnetic field inside was measured using a Hall probe placed into the holes on the printed structure. The measurements are found to be in good agreement with our finite element analysis results and indicate a fairly uniform field inside.

Effect of short and long term streptozotocin diabetes on smooth muscle calmodulin levels in the rat.

Diabetes mellitus is a metabolic disease associated with certain complications which have also been demonstrated in the experimental models of this disease. Altered responses to several agonists have been reported in various smooth muscles from alloxan or streptozotocin diabetic animals. Since these reports revealed a defect in the contractile process of smooth muscles from experimentally-induced diabetes, short and long term effects of diabetes on calmodulin levels in the smooth muscles of aorta, trachea, vas deferens and duodenum were investigated using streptozotocin diabetic rats. In spite of the fact that most of the reports have demonstrated the defective contractions in long term diabetic rats, short term effect (for 1 week) of diabetes on calmodulin levels in the smooth muscles of aorta, trachea, vas deferens and duodenum was also investigated in the present study using streptozotocin diabetic rats to understand whether the changes in calmodulin dependent contractile process begin at an earlier stage of the disease. Tissue calmodulin levels of the smooth muscles were measured by the radioimmunoassay technique using a [125I]-labeled kit. Although rats injected with streptozotocin exerted the characteristics of diabetes such as polyuria, polydipsy, polyphagy and elevated blood glucose levels, unchanged calmodulin levels were found in the rats with short term streptozotocin diabetes. In contrast, long term streptozotocin diabetes (for 8 weeks) was found to cause a significant decrease in tissue calmodulin levels of these four smooth muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Kinin receptors and their antagonists as novel therapeutic agents.

Plasma kinins are a family of peptides which play an important role in a variety of pathophysiological conditions. As a consequence of the progress in the area of kinin receptors, specific kinin receptor antagonists will be available in near future in order to provide more selective therapeutic modalities for the treatment of diseases such as asthma, cardiovascular diseases, inflammation, pain, etc. Although the initial observation leading to discovery of kinins has been performed in 1909, important progress on the development of specific kinin receptor antagonists has been made in last two decades. This review emphasizes the physiological functions of kinins along with their receptor subtypes and post-receptor events in the cellular signaling. In this article, the latest developments in the kinin receptor antagonists including nonpeptide ones have been reviewed.

Effect of insulin treatment on smooth muscle calmodulin levels in rats with long-term streptozotocin-diabetes.

Altered responses to several agonists have been reported in various smooth muscles from experimentally-diabetic animals suggesting a defective contractile process of smooth muscle. Recently, decreased smooth muscle calmodulin levels have been reported in streptozotocin-diabetic rats. However, the effectiveness of insulin on the decreased calmodulin levels in diabetic rats has not been questioned. Therefore, the present study was designed to examine the effect of insulin on smooth muscle calmodulin levels from streptozotocin-diabetic rats. Calmodulin levels of the smooth muscle were measured by a radioimmunoassay technique. Streptozotocin diabetes caused a significant decrease in tissue calmodulin levels of smooth muscles. Insulin therapy for 20 days did not correct the changes in calmodulin levels of rat smooth muscles, although it normalised blood glucose in streptozotocin-diabetic rats. These findings suggest that the altered smooth muscle calmodulin may contribute the defective contractile responses in diabetes and these changes may be resistant to insulin therapy.

Bradykinin receptors in isolated rat duodenum.

Pharmacological properties of the bradykinin receptors in the isolated rat duodenum were investigated by examining the relaxant and contractile responses to bradykinin and [des-Arg9]-bradykinin, an agonist of B1 receptors. A specific desensitization and de novo formation for B1 receptors were observed. Changes in medium pH caused a decrease in the responses to bradykinin and [des-Arg9]-bradykinin of rat duodenum. Urea incubation in test tube inhibited the responses to bradykinin and [des-Arg9]-bradykinin of rat duodenum, while urea in bathing medium was ineffective. These findings strongly suggested that (a) ionic bonds are important in the interaction between bradykinin and its receptors, and (b) B2 receptors in rat duodenum are different from those in guinea pig ileum.

Calmodulin content and in vitro contractility of duodenum from streptozotocin-induced diabetic rats: effects of insulin therapy and calmodulin antagonism.

The effects of experimental diabetes and insulin treatment on the decreased reactivity of isolated rat duodenum to KCl and calmidazolium, a specific calmodulin antagonist, were examined. After 8 weeks of streptozotocin diabetes, the contractile effect of KCl and the non-competitive antagonistic effect of calmidazolium against KCl on isolated rat duodenum were decreased. Calmodulin levels, as measured by radioimmunoassay, were also found to the decreased in duodenum from streptozotocin-diabetic rats. Neither impaired reactivity to KCl nor decreased calmodulin levels in diabetic rat duodenum were corrected by treatment with insulin (10 IU/kg for 20 days). Following insulin treatment, there was only a partial correction in the antagonistic effect of calmidazolium as shown by the increase in non-competitive antagonist affinity constant.

Acalculous cholecystitis: ascariasis as an unusual cause.

Migration of roundworms into the biliary tree is a well-known complication of Ascaris lumbricoides infestation of the intestine. Massive infestation of the hepato-biliary tract is uncommon but can lead to complications if not treated. Here, we report two cases of acalculous cholecystitis caused by ascariasis.

Decreased beta-adrenergic responses of rat small intestine due to non-insulin-dependent diabetes.

Relaxing responses to salbutamol of duodenum, jejunum and ileum isolated from alloxan-induced non-insulin-dependent diabetic rats were investigated by means of pharmacodynamic analysis using pD2 (apparent receptor affinity) and alpha E (intrinsic activity) values. beta-Adrenergic responses of these three intestinal parts of diabetic rats were found to be significantly decreased when compared to controls. The decreased responses may be attributable to a decrease in the number of beta-adrenergic receptors. The gastro-intestinal manifestations reported in diabetic patients are possibly linked to the occurrence of a decrease in beta-adrenergic receptors. Furthermore, this model seems to be useful for the investigations of gastro-intestinal complications resulting from non-insulin-dependent diabetes.

The effect of vanadate on alloxan-diabetic rat atria.

In this study, we examined the effect of vanadate treatment on cardiac changes recognized in diabetic rats. In addition, the possible contribution of thyroid hormones to vanadate's effect on alloxan-diabetic atria was also investigated. Administration of alloxan to rats, as expected, resulted in hyperglycemia; hypoinsulinemia reduced thyroid hormone levels, decreased body weight and depressed cardiac function. Vanadate treatment of diabetic rats normalized blood glucose and serum thyroid hormone levels, neither were serum insulin levels of diabetic animals corrected after vanadate treatment. Vanadate treatment, however, did not affect the body weights of diabetic rats. Spontaneously-beating atria from diabetic rats were found to have decreased rates but increased forces of contractions compared with those from controls. On the other hand, the responsiveness of diabetic atria to both inotropic and chronotropic effects of isoprenaline was found to be decreased. Vanadate treatment resulted in the normalization of these alterations observed in diabetic atria. These results thus indicate that the normalizing effect of vanadate on diabetes-induced hypothyroidism may contribute to its effect in preventing cardiac changes observed at the early stage of diabetes.

Duration-dependent changes in calcium responsiveness in the alloxan-diabetic rat intestine.

Disturbances of the gastrointestinal tract that are common in diabetes mellitus seem to be related to intestinal motility. In experimental models of diabetes, decreased calcium sensitivity has been demonstrated in various smooth muscles including those in the gastrointestinal tract. The main purpose of the present study was to examine further the calcium sensitivity in diabetic rat intestine and to understand if changes in the calcium sensitivity occur at an earlier stage of the disease. For this purpose, the effects of potassium and calcium were evaluated on nondepolarized and depolarized duodenum from rats with alloxan diabetes for 1 and 8 weeks and their age-matched controls. To evaluate the calcium sensitivity in rat duodenum, apparent affinity constants (pD2 values) and intrinsic activities (alpha E values) were calculated for every experimental conditions examined in this study. Both values (pD2 and alpha E) for the effects of potassium and calcium on the nondepolarized and depolarized duodenum, respectively, were not changed in 1-week diabetic rats. In contrast, intrinsic activities for the effects of potassium and calcium were found to be significantly decreased (p < 0.001) in the nondepolarized and depolarized duodenum from rats with alloxan diabetes for 8 weeks, whereas apparent affinity constants were not altered in this case. Taking into consideration all these experimental findings, the decreased calcium sensitivity in gastrointestinal tract seems to be closely related to decreased calmodulin levels and may occur at a later stage of diabetes as a linkage to long-term gastrointestinal complications.

Evaluation of contraction time and recovery period as a parameter in the calcium antagonistic action on the K(+)-depolarized rat duodenum.

Verapamil, nifedipine, phentolamine, tolazoline, gentamicin and neomycin inhibited calcium-induced contractions of K(+)-depolarized duodenum of rat by shifting the concentration-response curves to the right. Non-competitive inhibitions were observed with trifluoperazine, lidoflazine, procaine and tetracaine. Lanthanum behaved as a partial agonist in this preparation, while nitroprusside was ineffective. Contraction times in the presence of the antagonists and recovery time of the Ca2+ responses after the removal of the antagonists from the bathing medium were evaluated. From the findings, it is suggested that the contraction time and the time required for tissue recovery after removal of a Ca2+ antagonist are parameters making K(+)-depolarized rat duodenum a potential tool for the evaluation of the pharmacological effects of Ca2+ antagonists.

Nepetalactone: a new opioid analgesic from Nepeta caesarea Boiss.

The essential oils of Nepeta species including Nepeta phyllochlamys P. H. Davis, N. nuda L. ssp. nuda, and N. caesarea Boiss. have been screened by use of the tail-flick and tail immersion (52.5 degrees C) methods. Of the species studied, only N. caesarea showed significant analgesic activity, besides marked sedation, which was also blocked by naloxone, indicating involvement of opioid receptors. Moreover, it was only active on mechanical, not thermal, algesic response which suggests specificity for specific opioid receptor subtypes, excluding mu-opioid receptors. Because 4a alpha,7alpha,7a alpha-nepetalactone is the main component of the essential oil of N. caesarea, and is present at very high levels (92-95%), it is concluded that 4a alpha,7alpha,7a alpha-nepetalactone is the active principle and has a specific opioid receptor subtype agonistic activity.