Polymorphisms of CSF1 and TM7SF4 genes in a case of mild juvenile Paget’s disease found using next-generation sequencing

Juvenile Paget’s disease (JPD) is a rare autosomal-recessive condition. It is diagnosed in young children and characterized by a generalized increase in bone turnover, bone pain, and skeletal deformity. Our patient was diagnosed after a pathological fracture when she was 11 years old. When we first examined her at the age of 30 she had bone pain and deformity in both the femur and tibia. Serum alkaline phosphatase (ALP) level, radiology, bone scintigraphy, and densitometry were monitored. Next generation sequencing (NGS) technology, namely semiconductor sequencing, was used to determine the genetic background of JPD. Seven target genes and regions were selected and analyzed after literature review (TM7SF4, SQSTM1, TNFRSF11A, TNFRSF11B, OPTN, CSF1, VCP). No clear pathogenic mutation was found, but we detected missense polymorphisms in CSF1 and TM7SF4 genes. After treatment with zoledronic acid, infusion bone pain and ALP level decreased. We can conclude that intravenous zoledronic acid therapy is effective and safe for suppressing bone turnover and improving symptoms in JPD, but the long-term effects on clinical outcomes are unclear. Our findings also suggest that NGS may help explore the pathogenesis and aid the diagnosis of JPD.Juvenile Paget’s disease (JPD) is a rare genetic bone disease, with approximately 60 cases reported in total worldwide (1). In JPD, the function of the key proteins regulating osteoclast differentiation or function is affected. but like in Paget’s disease of bone (PDB), the exact pathomechanism is not understood. The genetic predisposition influences the function of bone cells. Though the gene candidates of the disease have not been identified yet, it is transmitted as an autosomal recessive trait (2,3). In most cases, there is a deficiency of the protein osteoprotegerin, leading to the clinical manifestations of the disease (3). No other affected genes have been identified yet.Some patients are asymptomatic, whereas others develop complications (4). JPD manifests in infancy or childhood, characterized by greatly accelerated and disorganized bone turnover (typically at focal areas), manifesting in bone deformities, pain secondary to fractures, osteopenia of the long bones, corticomedullary indistinctness, coarsening of the trabecular bone, growth retardation, deafness due to cochlear involvement, and nerve compression syndromes (2-4). Although clinically JPD has some similarities to PDB, the early age at onset and marked bone deformities can result in different pathomechanisms.We present a rare case of mild form of JPD with a genetic analysis – using a next generation sequencing technique – of seven target genes and regions of PDB. With the appearance of next-generation sequencing (NGS) machines, molecular biology reached a new revolutionary phase. This new technology combines high performance with much less expensive operation costs. Furthermore, we designed a novel approach to genetic testing in which we simultaneously sequenced the whole coding regions of the affected genes at the same time. Our approach was based on using an IonTorrent PGM from Life Technologies (Carlsbad, CA, USA). This benchtop sequencer belongs to the semiconductor sequencer family. It acquires the DNA sequence by detecting electric impulses created by the release of H⁺ -ions in its microchip. The solution that contains the H⁺-ions serves as a gate electrode of a transistor, a so called ion sensitive field electricity transistor (ISFET). This signal combined with cyclic addition of dNTP-s can be processed by the sequenator’s software as the incorporation of a nucleic acid, and several cycles will provide the sequence of the DNA in question. It is possible to barcode the samples with a sequence by which the software can differentiate between them. This is a great tool to sequence several samples at the same time, reducing the sequencing costs.We used the IonTorrent PGM (Life Technologies) to examine all genes known to play a role in the development of the different subtypes of Paget’s disease. Also, we treated our patient with zoledronate therapy, which makes this the second study to describe administration of such a medication in juvenile Paget’s disease (1).     

Multiple ABC Transporters Efflux Baicalin

Baicalein, the aglycone formed by hydrolysis of baicalin in the intestine, is well absorbed by passive diffusion but subjected to extensive intestinal glucuronidation. Efflux of baicalin, the low passive permeability glucuronide of baicalein from enterocytes, likely depends on a carrier‐mediated transport. The present study was designed to explore potential drug–herb interaction by investigating the inhibitory effect of baicalin on the transport of reporter substrates by transporters and to identify the transporters responsible for the efflux of baicalin from enterocytes and hepatocytes. The interaction of baicalin with specific ABC transporters was studied using membranes from cells overexpressing human BCRP, MDR1, MRP2, MRP3 and MRP4. Baicalin was tested for its potential to inhibit vesicular transport by these transporters. The transport of baicalin by the selected transporters was also investigated. Transport by BCRP, MRP3 and MRP4 was inhibited by baicalin with an IC₅₀ of 3.41 ± 1.83 μM, 14.01 ± 2.51 μM and 14.39 ± 5.69 μM respectively. Inhibition of MDR1 (IC₅₀ = 94.84 ± 31.10 μM) and MRP2 (IC₅₀ = 210.13 ± 110.49 μM) was less potent. MRP2 and BCRP are the apical transporters of baicalin that may mediate luminal efflux in enterocytes and biliary efflux in hepatocytes. The basolateral efflux of baicalin is likely mediated by MRP3 and MRP4 both in enterocytes and hepatocytes. Via inhibition of transport by ABC transporters, baicalin could interfere with the absorption and disposition of drugs. Copyright © 2015 John Wiley & Sons, Ltd.     

The 8.1 ancestral MHC haplotype is strongly associated with colorectal cancer risk

Many recent data indicate that some alleles encoded in the central major histocompatibility complex (MHC) region (Class III) of short arm of chromosome 6 may modify the risk of cancer development. Therefore we determined 4 single nucleotide polymorphisms (SNPs) of this region (TNF-alpha -308 G > A, RAGE -429 T > C, HSP70-2 -1267 A > G, LTA 252 A > G) in genomic DNA samples from 183 Hungarian patients with colorectal cancer and 141 age matched control subjects representing the Hungarian population of the same age and gender. No significant differences were found in either SNP tested. When, however, three- or four-locus haplotypes consisting of known constituents of the so-called 8.1 ancestral haplotype (8.1AH) were considered, marked differences were observed. Frequency of TNF-alpha -308A, RAGE -429C, HSP70-2 -1267G, LTA 252G (8.1AH) haplotype was significantly (p = 0.006) more frequent (19.1%) among patients than in the controls (7.7%). Age- and gender-adjusted ratio of the 8.1AH carriers vs. non-carriers to have colorectal cancer was 2.514 (1.130-5.594). This risk was higher in 相似文献   

Apolipoprotein A5 T-1131C variant confers risk for metabolic syndrome

The −1131C is a naturally occurring variant of the apolipoprotein A5 (ApoA5) gene, which has been shown to associate with increased triglyceride levels. This variant has also been shown to confer risk for development of ischemic heart disease and stroke. The gene is in linkage disequilibrium with factors known to correlate with impaired glucose homeostasis. These observations prompted us to study the prevalence of the ApoA5 –1131C allele in patients with metabolic syndrome. A total of 201 metabolic syndrome patients and 210 controls were studied. In both groups the triglyceride levels of patients with −1131C allele were significantly increased compared to the subjects with −1131T allele (3.22 ±0.43 mmol/1 vs. 2.24 ±0.12 mmol/1, p<0.01 in the metabolic syndrome patients; 2.10 ±0.19 mmol/1 vs. 1.22 ±0.05 mmol/1, p<0.01 in the controls). In metabolic syndrome patients the prevalence of the ApoA5 –1131C variant was increased compared to the healthy controls (11% vs. 6.20%). Multiplex regression analysis model adjusted for age, gender, serum total cholesterol levels, acute myocardial infarction and stroke events revealed that the examined ApoA5 variant confers risk for the development of metabolic syndrome: the odds ratio at 95% confidence interval was 3.622 (1.200–10.936), p=0.02. Our findings strongly suggest that this variant is a risk factor for the development of hypertriglyceridemia and metabolic syndrome.     

Insulin in flavin-containing monooxygenase regulation. Flavin-containing monooxygenase and cytochrome P450 activities in experimental diabetes.

Microsomal monooxygenases - cytochrome P450 (CYP, EC 1.14.14.1) and flavin-containing monooxygenase (FMO, EC 1.14.13.8) - have profound roles in drug metabolism. While the induction of some metabolic enzymes such as hepatic FMO and intestinal CYP1A, CYP2B is recognized in experimental diabetes, the effect of insulin treatment on FMO and intestinal CYP3A in diabetic animals has not been reported before. Changes in abundance and activity of hepatic and intestinal microsomal CYPs and FMO were studied in streptozotocin-induced diabetic rats either treated or not with insulin. Hepatic FMO1 activity increased in diabetic rats, but it was restored to control value on insulin treatment. Insulin itself had no effect on FMO1 activity in non-diabetic animals. A remarkable increase of total CYP content was accompanied by a reduced CYP3A specific enzyme activity in the small intestine of diabetic animals. The extent of these changes decreased on insulin treatment. Both, hepatic FMO1 and intestinal CYP3A activity correlated with average blood glucose concentration in untreated diabetic rats. These results indicate that insulin is involved in the regulation of hepatic FMO1 and intestinal CYP3A in rats. Blood glucose level is a good marker for FMO induction. The marked reduction of intestinal CYP3A capacity suggests that diabetes exerts a substantial effect on the activity of most determining intestinal CYP enzyme.     

Early recognition and successful treatment of an infant with severe combined immune deficiency

A male infant of three month presented with recurrent mucosal and fungal infections, diarrhoea and failure to thrive from the age of three weeks. Laboratory test revealed T-B-NK + severe combined immunodeficiency (SCID). Family history and immunolaboratory findings suggested autosomal recessive form of the disease. Haploidentical maternal bone marrow transplantation (BMT) was carried out at five and half months of age. Over the two years after BMT, the patient's somatomotoric and mental development is normal. Cellular immune responses and the substantial immunoglobulin production suggest immunoreconstruction in the child born with complete lack of adaptive immunity. According to the author's knowledge, this is the first T-B-NK + patient in Hungary, whose disease was diagnosed and adequately treated in infancy.     

Value of rilmenidine therapy and its combination with perindopril on blood pressure and left ventricular hypertrophy in patients with essential hypertension (VERITAS)

OBJECTIVES: The primary objective was to assess the effects of rilmenidine monotherapy and in combination with perindopril on blood pressure (BP) in patients assessed with grade 1 or 2 essential hypertension. The study also examined the effects of 2-year rilmenidine monotherapy on left ventricular hypertrophy (LVH) and on diastolic function of the left ventricle, along with the effects of rilmenidine on left ventricular mass index in hypertensive patients with no LVH, and the relationship between BP reduction and any change in LVH. RESEARCH DESIGN AND METHODS: Mild-to-moderate hypertensive patients (n = 500) were enrolled in a multicentre 2-year open study and treated with rilmenidine (1-2 mg per day) monotherapy or rilmenidine plus perindopril (2, 4 or 8 mg per day) if control of hypertension was not achieved with rilmenidine monotherapy within 12 weeks. Blood pressure was recorded at regular intervals by the investigators and LVH measured by centralised single-blind echocardiographic reading. RESULTS: Rilmenidine monotherapy (average dose 1.42 mg) produced a significant decrease in BP from the baseline of 163 +/- 10/100 +/- 5 mmHg to 134 +/- 10/86 +/- 7 mmHg at 1 year and to 136 +/- 10/84 +/- 7 mmHg at 2 years (p < 0.001 for both). In 188 patients with LVH, the left ventricular mass index was significantly reduced from 161.4 +/- 30.5 to 131.3 +/- 26.5 at 1 year and to 134.1 +/- 26.0 g/m(2) at 2 years (p < 0.001 for both). Addition of perindopril to those patients whose BP was not normalised by rilmenidine monotherapy after 12 weeks further decreased BP significantly from 150 +/- 13/93 +/- 8 mmHg to 142 +/- 14/89 +/- 7 mmHg at the end of the 2nd year. CONCLUSIONS: Long-term rilmenidine monotherapy was shown to be efficient in controlling BP and in reducing LVH. The addition of perindopril to rilmenidine monotherapy proved to be effective and well tolerated in those patients who did not respond to rilmenidine alone.     

Effect of sustained low efficient dialysis versus continuous renal replacement therapy on renal recovery after acute kidney injury in the intensive care unit: A systematic review and meta‐analysis

Critically ill adults with acute kidney injury (AKI) experience considerable morbidity and mortality. Controversy remains regarding the optimal renal replacement intervention for these patients. Our systematic review aimed to determine the effect(s) of sustained low‐efficiency dialysis (SLED) compared with continuous renal replacement (CRRT) therapy on relevant patient outcomes. A systematic search of Medline, Embase, CINAHL and the Cochrane Library was conducted. Identified citations were screened independently in duplicate for relevance, and the methodological quality of included studies was evaluated. Data were extracted on study, patient and intervention characteristics and relevant clinical outcomes. Results were pooled using inverse variance fixed and random effects meta‐analysis. A total of 1564 patients from 18 studies were included. Meta‐analysis results indicated no statistically significant difference in our primary outcome, overall proportion of renal recovery (risk ratio (RR) 0.87, 95% confidence interval (CI) 0.63–1.20, I2 = 66%). No significant difference was observed for the secondary outcome of time to renal recovery (mean difference 1.33, 95% CI 0.23–2.88, I2 = 0%). Statistically, SLED was marginally favoured over CRRT for the secondary outcome of mortality (RR 1.21, 95% CI 1.02–1.43, I2 = 47%); however, this diminished when sensitivity analysis of only randomized controlled trials was conducted (RR 1.25, 95% CI 1.00–1.57, I2 = 0%). There appears to be no clear for advantage continuous renal replacement in the hemodynamically unstable patient. Currently, both modalities are safe and effective means of treating AKI in the critically ill adult.