Osteonecrosis of the jaw among cancer patients in Denmark: risk and prognosis.

Osteonecrosis of the jaw (ONJ) is a serious complication of anti-resorptive therapy used in the treatment of multiple myeloma and cancerous bone metastases. In this study, patients with either multiple myeloma or solid tumours with a simultaneous or subsequent record of anti-resorptive treatment or bone metastases were identified using population-based medical registries. These patients were followed for the outcome of ONJ. Considering death as a competing risk, the cumulative incidence of ONJ was estimated, overall and by cancer site. Patients who developed ONJ were followed for the outcome of death overall and by several risk factors for ONJ. A total of 33,975 cancer patients fulfilling the inclusion criteria were identified; 233 incidents of ONJ and a cumulative incidence of 1.9% (95% confidence interval 1.6-2.3%) over a maximum follow-up time of 7.5 years were observed. The 5-year cumulative incidence was 1.3% (95% confidence interval 1.2-1.6%) and varied by cancer site. There were 126 deaths among cancer patients with ONJ over a maximum follow-up time of 6.4 years, resulting in a 5-year mortality of 91% (95% confidence interval 81-97%). Mortality among patients with ONJ varied by cancer site, osteonecrosis stage, and by history of trauma to the mucosa.

Genetically determined high activity of IL-12 and IL-18 in ulcerative colitis and TLR5 in Crohns disease were associated with non-response to anti-TNF therapy.

Anti-tumour necrosis factor-α (TNF-α) is used for treatment of severe cases of inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC). However, one-third of the patients do not respond to the treatment. A recent study indicated that genetically determined high activity of pro-inflammatory cytokines, including interleukin-1ß (IL-1ß), IL-6 and interferon gamma (IFN-γ), are associated with non-response to anti-TNF therapy. Using a candidate gene approach, 21 functional single-nucleotide polymorphisms (SNPs) in 14 genes in the Toll-like receptors, the inflammasome and the IFNG pathways were assessed in 482 and 256 prior anti-TNF naïve Danish patients with CD and UC, respectively. The results were analysed using logistic regression (adjusted for age and gender). Eight functional SNPs were associated with anti-TNF response either among patients with CD (TLR5 (rs5744174) and IFNGR2 (rs8126756)), UC (IL12B (rs3212217), IL18 (rs1946518), IFNGR1 (rs2234711), TBX21 (rs17250932) and JAK2 (rs12343867)) or in the combined cohort of patient with CD and UC (IBD) (NLRP3 (rs10754558), IL12B (rs3212217) and IFNGR1 (rs2234711)) (P<0.05). Only the association with heterozygous genotype of IL12B (rs3212217) (OR: 0.24, 95% CI: 0.11-0.53, P=0.008) among patients with UC withstood Bonferroni correction for multiple testing. In conclusion, Our results suggest that SNPs associated with genetically determined high activity of TLR5 among patients with CD and genetically determined high IL-12 and IL-18 levels among patients with UC were associated with non-response. Further studies will evaluate whether these genes may help stratifying patients according to the expected response to anti-TNF treatment.

Complete sequence-based pathway analysis by differential on-chip DNA and RNA extraction from a single cell.

We demonstrate on-chip, differential DNA and RNA extraction from a single cell using a microfluidic chip and a two-stage lysis protocol. This method enables direct use of the whole extract, without additional washing steps, reducing sample loss. Using this method, the tumor driving pathway in individual cells from a colorectal cancer cell line was determined by applying a Bayesian computational pathway model to sequences obtained from the RNA fraction of a single cell and, the mutations driving the pathway were determined by analyzing sequences obtained from the DNA fraction of the same single cell. This combined functional and mutational pathway assessment of a single cell could be of significant value for dissecting cellular heterogeneity in tumors and analyzing single circulating tumor cells.

Associations between functional polymorphisms in the NFκB signaling pathway and response to anti-TNF treatment in Danish patients with inflammatory bowel disease.

Antitumor necrosis factor-α (TNF-α) is used for treatment of severe cases of inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC). However, one-third of the patients do not respond to the treatment. Genetic markers may predict individual response to anti-TNF therapy. Using a candidate gene approach, 39 mainly functional single nucleotide polymorphisms (SNPs) in 26 genes regulating inflammation were assessed in 738 prior anti-TNF-naive Danish patients with IBD. The results were analyzed using logistic regression (crude and adjusted for age, gender and smoking status). Nineteen functional polymorphisms that alter the NFκB-mediated inflammatory response (TLR2 (rs3804099, rs11938228, rs1816702, rs4696480), TLR4 (rs5030728, rs1554973), TLR9 (rs187084, rs352139), LY96 (MD-2) (rs11465996), CD14 (rs2569190), MAP3K14 (NIK) (rs7222094)), TNF-α signaling (TNFA (TNF-α) (rs361525), TNFRSF1A (TNFR1) (rs4149570), TNFAIP3(A20) (rs6927172)) and other cytokines regulated by NFκB (IL1B (rs4848306), IL1RN (rs4251961), IL6 (rs10499563), IL17A (rs2275913), IFNG (rs2430561)) were associated with response to anti-TNF therapy among patients with CD, UC or both CD and UC (P ⩽ 0.05). In conclusion, the results suggest that polymorphisms in genes involved in activating NFκB through the Toll-like receptor (TLR) pathways, genes regulating TNF-α signaling and cytokines regulated by NFκB are important predictors for the response to anti-TNF therapy among patients with IBD. Genetically strong TNF-mediated inflammatory response was associated with beneficial response. In addition, the cytokines IL-1ß, IL-6 and IFN-γ may be potential targets for treating patients with IBD who do not respond to anti-TNF therapy. These findings should be examined in independent cohorts before these results are applied in a clinical setting.

Efficacy and safety of androgen deprivation therapy after switching from monthly leuprolide to monthly degarelix in patients with prostate cancer.

OBJECTIVES: To evaluate whether switching prostate cancer (PCa) patients from leuprolide to degarelix is associated with any change in the efficacy of testosterone suppression or safety profile during the first 3 months. METHODS: Participants were 134 patients with histologically confirmed PCa who had completed 1 year of treatment with leuprolide 7.5 mg monthly before being switched to degarelix. These patients were re-randomised for the extension trial to receive a starting dose of 240 mg degarelix followed by monthly maintenance doses of either 80 (n = 69) or 160 mg (n = 65). For efficacy assessment, serum testosterone, prostate-specific antigen (PSA), luteinising hormone (LH) and follicle-stimulating hormone (FSH) levels measured at days 3, 7, 14, 28, 56 and 84 assessed whether treatment efficacy is sustained. Safety and tolerability assessments included adverse events (AEs), physical examinations, electrocardiograms and clinically significant changes in laboratory safety parameters. RESULTS: Serum testosterone, LH, and PSA levels were all sustained in both treatment arms during the observation period. Interestingly, FSH levels were further decreased by 30% following the switch to degarelix. With the exception of injection site reactions, the overall prevalence and pattern of AEs during the first 3 months after the switch was comparable to that during the last 3 months leuprolide treatment in the main trial. There were five (4%) patients discontinued to treatment-related AEs including injection site pain (n = 3) and fatigue (n = 2). CONCLUSIONS: This 3-month analysis indicates that patients with prostate cancer can be safely switched from leuprolide to degarelix treatment with sustained efficacy as measured by biochemical markers.

Degradation of 4-nonylphenol in homogeneous and nonhomogeneous mixtures of soil and sewage sludge.

Sewage sludge is frequently applied as fertilizers to cultivated land. However, municipal sewage sludge often contains organic contaminants including nonylphenol (NP), an intermediate from nonionic surfactant degradation. Knowledge about NP degradation in sludge-amended soil is an important prerequisite for adequate risk assessments. In this study, mineralization of 14C-labeled NP in homogenized and nonhomogenized sludge-soil mixtures was investigated. NP was degraded within 38 days in aerobic homogenized mixtures. In nonhomogeneous mixtures containing sludge aggregates, the degradation of NP was retarded and was generally not completed within 3 months (119-126 days). No detectable amounts of NP were transported from the sludge aggregates to the surrounding soil (detection limit: <0.04 mg of NP/kg dw of soil). Oxygen penetration into sludge aggregates was monitored for 50 days with an oxygen microelectrode. An extrapolation of the oxygen data suggested that more than 1 year was required to obtain fully aerobic conditions in a 2-cm sludge aggregate. Since NP is considered persistent in the absence of oxygen, residual amounts of NP may be present in the anaerobic center of aggregates for prolonged periods. The results demonstrate that sludge aggregate size and thus oxygen availability will be a major controlling factor for NP degradation in soil amended with sewage sludge and that the mobility of NP from sludge aggregates to the surrounding soil is negligible.

[Spontaneous rectal hematoma]. / Spontant rectushaematom.

Five cases of spontaneous rectus sheath haematomas observed during one year are reported. The female to male ratio was four to one. The age range was 26-75 years. Precipitating factors were cough in three patients, physical exercise in one patient, and none in one patient. In three cases there were possible predisposing factors: anticoagulant therapy, Addison's disease with steroid treatment and abdominal scar. The localisations were three lower right quadrant, one lower left and one upper right. In three cases correct diagnoses were made based on clinical findings, confirmed by ultrasonography and treated conservatively. In two cases the diagnosis was not suspected clinically, and by ultrasonography the haematoma was overlooked in one case, and misinterpreted as an intraperitoneal tumour in the other; both cases underwent surgery. The incidence rate was estimated to 3.5 per 100,000 per year. The frequency was approximately 0.9% in patients admitted with "acute abdomen".

[Profile of pediatric diseases and symptoms in Southern Greenland. Experiences of a consultant visiting in 1992]. / Paediatrisk sygdoms- og symptomprofil i Sydgrønland. Erfaringer fra konsulentbesøg i 1992.

During a scheduled visit to southern Greenland in 1992, the pediatric consultant registered signs and symptoms that had led to the children being brought for examination, and simultaneously registered the actual frequency of asthma, epilepsia and cerebral palsy. A total of 211 children or 8% of the background population under the age of fifteen were examined. The point prevalence of asthma was 1.1% which was considered to be an absolute minimum value due to selection of patients. The point of epilepsy was 0.6% and cerebral palsy 0.3%, a surprisingly high prevalence compared to prior investigations and actual patient selection. The consultant was met as well by a broad spectrum of pediatric problems, many of the social pediatric type. In conclusion, the importance of yearly visits is maintained along with a need of instruction of the local local health personnel. A systematic registration of cerebral palsy cases is required in order to implement necessary prophylactic intervention. Contact between the Greenlandic and the Danish health services could be intensified by the use of electronic media.

Physiological responses of Atherosperma moschatum to day length, night length and photosynthetic photon fluence rate.

Young, non-reproductive plants of Atherosperma moschatum Labill, were conditioned to a range of day lengths, night lengths and photosynthetic photon fluence rates (PPFRs). The plants conditioned to the shorter of the two day lengths grew more slowly. On protein specific bases, the leaves of these plants had greater PPFR-saturated CO2 exchange rates (CERs), steeper initial slopes to the CER-PPFR plots, and greater chlorophyll concentrations than did the leaves of the long day length plants. The rates of leaf respiration, and the rates of leaf carbon export and /or incorporation into structural leaf products (E+ I) were similar under the two day lengths. The plants conditioned to the shorter of the two night lengths grew more quickly. The leaves ot these plants had smaller PPFR-saturated CERs, shallower initial slopes to the CER-PPFR plots and smaller chlorophyll concentrations than did the leaves of the long night length plants, but greater rates of leaf respiration and E + I. In a separate experiment A. moschatum was shown to be vegetatively sensitive to night breaks of far-red irradiation. The plants conditioned to the higher of the two PPFRs grew more quickly. The leaves of these plants had greater PPFR-saturated CERs, greater rates of leaf respiration, and greater rates of E+I than did the leaves of the lower PPFR plants, but shallower initial slopes to the CER-PPFR plots, and smaller chlorophyll concentrations. Acclimation to photosynthetic period (day length/night length) can be as marked as that to PPFR, and is quite different in character.

A common response element mediates differential effects of phorbol esters and forskolin on type-1 plasminogen activator inhibitor gene expression in human breast carcinoma cells.

We have characterized regulation of type-1 plasminogen activator inhibitor (PAI-1) gene expression by phorbol 12-myristate 13-acetate (PMA) and the cAMP-inducing agent forskolin in the human breast carcinoma cell line MCF-7. PMA caused a strong induction of PAI-1, while forskolin suppressed the PMA response. Transfection experiments with fusion genes showed that sequences mediating PMA induction as well as forskolin suppression were present between base pairs -100 and -30 of the 5'-flanking region of the PAI-1 gene. The region was found to contain two Sp1 binding sites. A proximal sequence in the region, TGAGTTCA (P box), with sequence similarity to phorbol ester response elements (TRE) as well as to cAMP response elements (CRE), bound a low-abundance, as yet unidentified nuclear protein in MCF-7 cells. This sequence had a higher affinity to purified c-jun homodimer than to c-jun/c-fos heterodimer in MCF-7 nuclear extracts; it had no affinity to the proteins binding to CRE consensus sequences in these extracts. A distal TRE-like sequence, TGAGTGG (D box), had a weak affinity to c-jun/c-fos heterodimer and c-jun homodimer; binding of proteins to this sequence was facilitated by binding of proteins to the P box. Both the P box and the D box were necessary for PMA responsiveness, suggesting a cooperativity between the two binding sites. A mutation of the P box removing the CRE similarity abolished the forskolin suppression of the PMA response. We propose that the protein kinase C and the protein kinase A signal-transduction pathways, with opposite effects on PAI-1 gene expression converge by modulating differently P-box-binding proteins.

Transforming growth factor beta 1-responsive element: closely associated binding sites for USF and CCAAT-binding transcription factor-nuclear factor I in the type 1 plasminogen activator inhibitor gene.

Transforming growth factor beta (TGF-beta) is the name of a group of closely related polypeptides characterized by a multiplicity of effects, including regulation of extracellular proteolysis and turnover of the extracellular matrix. Its cellular mechanism of action is largely unknown. TGF-beta 1 is a strong and fast inducer of type 1 plasminogen activator inhibitor gene transcription. We have identified a TGF-beta 1-responsive element in the 5'-flanking region of the human type 1 plasminogen activator inhibitor gene and shown that it is functional both in its natural context and when fused to a heterologous nonresponsive promoter. Footprinting and gel retardation experiments showed that two different nuclear factors, present in extracts from both TGF-beta 1-treated and nontreated cells, bind to adjacent sequences contained in the responsive unit. A palindromic sequence binds a trans-acting factor(s) of the CCAAT-binding transcription factor-nuclear factor I family. A partially overlapping dyad symmetry interacts with a second protein that much evidence indicates to be USF. USF is a transactivator belonging to the basic helix-loop-helix family of transcription factors. Mutations which abolish the binding of either CCAAT-binding transcription factor-nuclear factor I or USF result in reduction of transcriptional activation upon exposure to TGF-beta 1, thus showing that both elements of the unit are necessary for the TGF-beta 1 response. We discuss the possible relationship of these findings to the complexity of the TGF-beta action.

Effect of near normoglycemia for 5 years on progression of early diabetic retinopathy and renal involvement.

The effect of strict metabolic control for 5 years on renal function and retinal morphology was estimated in 24 insulin-dependent diabetic individuals (age 29 +/- 8 years, diabetes duration 10 +/- 6 years) with albustix negative urine and minimal or no background retinopathy before the study. They were randomized to conventional insulin treatment (CIT) or continuous subcutaneous insulin infusion (CSII) with a portable pump. During CSII treatment the metabolic status was significantly improved. HbA1c fell from 8.9 +/- 2.0 to 7.4 +/- 1.3% (p less than 0.01) whereas HbA1c was unchanged during CIT treatment. The mean value of urinary albumin excretion (UAE) was not statistically significantly changed (from 12 +/- 10 mg/24 h to 13 +/- 5 mg/24 h in CSII patients and from 14 +/- 12 to 11 +/- 6 mg/24 h in CIT patients (p greater than 0.1). On the other hand, the elevated GFR values were significantly reduced in both CSII and CIT patients, 129 +/- 17 to 120 +/- 9 and 129 +/- 18 to 119 +/- 12 ml/min.1,73m2, respectively (p less than 0.05). Both the number of microaneurysms, haemorrhages and exudates tended to increase mor in CIT patients than in CSII patients during the 5 year study period, but the differences did not reach statistically significance (p greater than 0.1). Pump treatment did not induce proliferations or "cotton wool" exudates. We conclude that no statistically significantly differences between GFR values, UAE rates, and progression of background retinopathy was observed between normoalbuminuric IDDM patients treated for 5 years with CIT and CSII, respectively. However, due to the size of the material a type II error must be taken into consideration.

A unique tRNA intron in the variable loop of the extreme thermophile Thermofilum pendens and its possible evolutionary implications.

We describe, for the first time, an intron that is located in the variable loop of a tRNA. It is 18 nucleotides long and occurs within a precursor tRNAGly from the extreme thermophile Thermofilum pendens. The variable loop is less critical for tRNA function than the anticodon arm where other class III introns occur. This raises the possibility that tRNAs with a large variable loop (type II) arose from those with a small variable loop (type I) by retaining a splicing-deficient intron.

Comparison of transfer RNA and ribosomal RNA intron splicing in the extreme thermophile and archaebacterium Desulfurococcus mobilis.

The structure of the exon-intron boundary was compared for an intron within 23S ribosomal RNA of Desulfurococcus mobilis and a newly discovered intron in tRNA(Met) from the same organism. The occurrence of a putative common structural feature suggests that intron excision occurs by the same mechanism. The possible recognition of this structural feature by the cleavage enzyme was investigated for the ribosomal RNA intron using RNA substrates exhibiting various exon and intron deletions. The results support the involvement of the structural features in the cleavage process. The evolutionary implications of these results are considered.

Somatic manifestations in children suspected of having been maltreated.

The present study revealed an incidence of maltreatment of 26.8 in 100,000 children under 15 years of age, higher than previously assumed. The sample comprises 41 children suspected of having been maltreated. A systematic examination included radiography and isotope scintigraphy of the skeletal system. In 9 cases the diagnosis was not confirmed, in 23 confirmed, and in 9 children it seemed likely. The most common form of maltreatment was physical violence with bruises. A total of 6 fractures were demonstrated. Three children had been exposed to sexual abuse. Intracranial haematoma, of long standing, was found in one child. Thirteen of the children (40%) were of short statute, their height and weight below the 25th percentile. Among 27 children examined by X-rays 16 (59%) had delayed skeletal maturation. Disturbances of growth, as well as behavioural abnormalities, were particularly striking in this group of patients. The findings suggest that even the slightest suspicion of maltreatment should lead to a general medical evaluation without delay.