The ESMO Tumour-Agnostic Classifier and Screener (ETAC-S): a tool for assessing tumour-agnostic potential of molecularly guided therapies and for steering drug development.

BACKGROUND: Advances in precision oncology led to approval of tumour-agnostic molecularly guided treatment options (MGTOs). The minimum requirements for claiming tumour-agnostic potential remain elusive. METHODS: The European Society for Medical Oncology (ESMO) Precision Medicine Working Group (PMWG) coordinated a project to optimise tumour-agnostic drug development. International experts examined and summarised the publicly available data used for regulatory assessment of the tumour-agnostic indications approved by the US Food and Drug Administration and/or the European Medicines Agency as of December 2023. Different scenarios of minimum objective response rate (ORR), number of tumour types investigated, and number of evaluable patients per tumour type were assessed for developing a screening tool for tumour-agnostic potential. This tool was tested using the tumour-agnostic indications approved during the first half of 2024. A taxonomy for MGTOs and a framework for tumour-agnostic drug development were conceptualised. RESULTS: Each tumour-agnostic indication had data establishing objective response in at least one out of five patients (ORR ≥ 20%) in two-thirds (≥4) of the investigated tumour types, with at least five evaluable patients in each tumour type. These minimum requirements were met by tested indications and may serve as a screening tool for tumour-agnostic potential, requiring further validation. We propose a conceptual taxonomy classifying MGTOs based on the therapeutic effect obtained by targeting a driver molecular aberration across tumours and its modulation by tumour-specific biology: tumour-agnostic, tumour-modulated, or tumour-restricted. The presence of biology-informed mechanistic rationale, early regulatory advice, and adequate trial design demonstrating signs of biology-driven tumour-agnostic activity, followed by confirmatory evidence, should be the principles for tumour-agnostic drug development. CONCLUSION: The ESMO Tumour-Agnostic Classifier (ETAC) focuses on the interplay of targeted driver molecular aberration and tumour-specific biology modulating the therapeutic effect of MGTOs. We propose minimum requirements to screen for tumour-agnostic potential (ETAC-S) as part of tumour-agnostic drug development. Definition of ETAC cut-offs is warranted.

Use of monofilament sutures and triclosan coating to protect against surgical site infections in spinal surgery: a laboratory-based study.

PURPOSE: We investigated bacterial propagation through multifilament, monofilament sutures and whether sutures coated with triclosan would exhibit a different phenomenon. METHODS: One centimetre (cm) wide trenches were cut in the middle of Columbia blood Agar plates. We tested a 6 cm length of two Triclosan-coated (PDS plus®, Vicryl plus®) and two uncoated (PDS ®, Vicryl ®) sutures. Each suture was inoculated with a bacterial suspension containing methicillin-sensitive Staphylococcus aureus (MSSA), Escherichia coli (E. coli), Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus (MRSA) at one end of each suture. The plates were incubated at 36C for 48 h, followed by room temperature for a further 5 days. We established bacterial propagation by observing for any bacterial growth on the Agar on the opposite side of the trench. RESULTS: Bacterial propagation was observed on the opposite side of the trench with both suture types, monofilament PDS and multifilament Vicryl, when tested with the motile bacterium (E. coli). Propagation was not observed on the other side of the trench with the monofilament PDS suture following incubation with MSSA and S. epidermidis, and in 66% of MRSA. With multifilament suture Vicryl, propagation was observed on the other side of the trench in 90% (MSSA), 80% (S. epidermidis), and 100% (MRSA) of plates tested. No bacterial propagation was observed in any of the triclosan-coated sutures (monofilament or multifilament). CONCLUSIONS: Monofilament sutures are associated in vitro with less bacterial propagation along their course when compared to multifilament sutures. Inhibition in both sutures can be further enhanced with a triclosan coating.

Beyond epidermal growth factor receptor: MET amplification as a general resistance driver to targeted therapy in oncogene-driven non-small-cell lung cancer.

The rapidly changing treatment paradigm for patients with metastatic oncogene-driven lung cancer continues to evolve, and consequently our understanding of the landscape of resistance must also advance. MET amplification is an established and frequent driver of resistance in EGFR-mutant non-small-cell lung cancer (NSCLC). Recently, the combination of MET proto-oncogene (MET) and epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) has shown promise in overcoming this molecularly defined resistance in clinical trials, and this combination strategy is being pursued in ongoing trials. Emerging data also demonstrate MET amplification as a resistance driver to TKI-treated ALK-, RET-, and ROS-1-fusion NSCLC, consistently at the range of 15%, while the resistance profiling data are maturing for other molecular targets. In this review, we discuss MET amplification as a driver of acquired resistance in well-defined molecular subsets of NSCLC, explore the biology behind this mechanism of resistance, and summarize the recently published clinical data, including the proposed combination strategies in the clinic achieving success in overcoming acquired MET amplification-dependent resistance.

Parish Prevalence Estimates of Obesity, Diabetes and Hypertension in Jamaica: Is It Now Time for More Targeted Public Health Interventions? / Estimados de la Prevalencia de la Obesidad, la Diabetes y la Hipertensión por Parroquia en Jamaica: ¿Es ya Tiempo de Realizar Intervenciones de Salud Pública Más Específicas?

ABSTRACT Objective: Geographic variation in obesity, Diabetes mellitus (DM) and hypertension (HTN) prevalence at the parish level was examined using the Jamaica Health and Lifestyle Survey 2008 (JHLS II). Methods: Total and sex-specific parish age-adjusted prevalence estimates of obesity, DM and HTN were obtained and ranked. Binary logistic regression models were adjusted for age, urbanicity, educational level, physical activity and diet. Results: Parish prevalence ranges were obesity 19.5-37.8% (1.7-31.0% in men versus 27.39-48.30% in women); DM 5.08-37.82% (0-26.45% in men versus 7.11-14.17% in women) and HTN 19.50-36.02% (10.94-48.39% in men versus 18.85-36.61% in women). The highest parish prevalences were St Elizabeth for obesity, Portland for DM and St Mary for HTN. Men residing in St Elizabeth were 16 times more obese compared to those in Portland [(Odds Ratio) OR = 15.84; 95% CI = 2.00, 125.51, p < 0.01], while women in St Elizabeth had twice the odds of being obese compared to those in St Ann [OR = 2.3; 95% CI, 1.007, 5.3). Men in Portland were eight times more likely to have HTN compared to those residing in St Ann (OR = 7.70; 95% CI = 2.34, 25.40, p = 0.001) whilst women in St Mary were three times more likely to be hypertensive compared to those living in St Thomas (OR = 3.05; 95% CI = 1.63, 5.72, p = 0.001). No significant associations were seen with DM. Conclusion: Significant heterogeneity exists at the parish level in obesity, DM and HTN, with important sex differences. Further analyses are needed to understand the determinants and work toward context-specific prevention and intervention programming.

RESUMEN Objetivo: La variación geográfica de la prevalencia de la obesidad, la diabetes mellitus (DM) y la hipertensión (HT) a nivel parroquia, se examinó usando la Encuesta 2008 sobre Salud y Estilo de Vida de Jamaica (JHLS-2). Métodos: Los estimados totales y específicos por género, ajustados por edad y a nivel de parroquia, de la prevalencia de la obesidad, DM y HT, fueron obtenidos y clasificados. Los modelos de regresión logística binaria fueron ajustados por edad, urbanidad, nivel educacional, actividad física, y dieta. Resultados: Los rangos de prevalencia por parroquia fueron como sigue: obesidad 19.5- 37.8% (1.7-31.0% en hombres versus 27.39-48.30% en mujeres); DM 5.08-37.82% (0- 26.45% en hombres versus 7.11-14.17% en mujeres); y HT 19.50-36.02% (10.94-48.39% en hombres versus 18.85-36.61% en mujeres). Las prevalencias más altas por parroquia fueron: Saint Elizabeth en obesidad, Portland en DM, y Saint Mary en HT. Los hombres de Saint Elizabeth eran 16 veces más obesos en comparación con los de Portland [(Odds Ratio) OR = 15.84; 95% IC = 2.00, 125.51, p < 0.01], mientras que las mujeres de Saint Elizabeth tenían el doble de probabilidades de ser obesas en comparación con las de Saint Ann (OR = 2.3; 95% IC, 1.007, 5.3). Los hombres de Portland eran ocho veces más propensos a padecer de HT en comparación con los residentes en Saint Ann (OR = 7.70; 95% IC = 2.34, 25.40, p = 0.001) en tanto que las mujeres de Saint Mary tenían tres veces más probabilidades de ser hipertensas comparadas con las que viven en Saint Thomas (OR = 3.05; 95% IC = 1.63, 5.72, p = 0.001). No se observaron asociaciones significativas con DM. Conclusión: Existe una heterogeneidad significativa a nivel de parroquias en cuanto a obesidad, DM, y HT, con importantes diferencias de género. Se necesitan más análisis para entender las determinantes y trabajar hacia la programación de intervenciones y prevenciones específicas del contexto.

HPV16 oncogene expression levels during early cervical carcinogenesis are determined by the balance of epigenetic chromatin modifications at the integrated virus genome.

In cervical squamous cell carcinomas, high-risk human papillomavirus (HRHPV) DNA is usually integrated into host chromosomes. Multiple integration events are thought to be present within the cells of a polyclonal premalignant lesion and the features that underpin clonal selection of one particular integrant remain poorly understood. We previously used the W12 model system to generate a panel of cervical keratinocyte clones, derived from cells of a low-grade premalignant lesion naturally infected with the major HRHPV type, HPV16. The cells were isolated regardless of their selective advantage and differed only by the site of HPV16 integration into the host genome. We used this resource to test the hypothesis that levels of HPV16 E6/E7 oncogene expression in premalignant cells are regulated epigenetically. We performed a comprehensive analysis of the epigenetic landscape of the integrated HPV16 DNA in selected clones, in which levels of virus oncogene expression per DNA template varied ~6.6-fold. Across the cells examined, higher levels of virus expression per template were associated with more open chromatin at the HPV16 long control region, together with greater loading of chromatin remodelling enzymes and lower nucleosome occupancy. There were higher levels of histone post-translational modification hallmarks of transcriptionally active chromatin and lower levels of repressive hallmarks. There was greater abundance of the active/elongating form of the RNA polymerase-II enzyme (RNAPII-Ser2P), together with CDK9, the component of positive transcription elongation factor b complex responsible for Ser2 phosphorylation. The changes observed were functionally significant, as cells with higher HPV16 expression per template showed greater sensitivity to depletion and/or inhibition of histone acetyltransferases and CDK9 and less sensitivity to histone deacetylase inhibition. We conclude that virus gene expression per template following HPV16 integration is determined through multiple layers of epigenetic regulation, which are likely to contribute to selection of individual cells during cervical carcinogenesis.

Biology of childhood germ cell tumours, focussing on the significance of microRNAs.

Genomic and protein-coding transcriptomic data have suggested that germ cell tumours (GCTs) of childhood are biologically distinct from those of adulthood. Global messenger RNA profiles segregate malignant GCTs primarily by histology, but then also by age, with numerous transcripts showing age-related differential expression. Such differences are likely to account for the heterogeneous clinico-pathological behaviour of paediatric and adult malignant GCTs. In contrast, as global microRNA signatures of human tumours reflect their developmental lineage, we hypothesized that microRNA profiles would identify common biological abnormalities in all malignant GCTs owing to their presumed shared origin from primordial germ cells. MicroRNAs are short, non-protein-coding RNAs that regulate gene expression via translational repression and/or mRNA degradation. We showed that all malignant GCTs over-express the miR-371-373 and miR-302/367 clusters, regardless of patient age, histological subtype or anatomical tumour site. Furthermore, bioinformatic approaches and subsequent Gene Ontology analysis revealed that these two over-expressed microRNAs clusters co-ordinately down-regulated genes involved in biologically significant pathways in malignant GCTs. The translational potential of this finding has been demonstrated with the detection of elevated serum levels of miR-371-373 and miR-302/367 microRNAs at the time of malignant GCT diagnosis, with levels falling after treatment. The tumour-suppressor let-7 microRNA family has also been shown to be universally down-regulated in malignant GCTs, because of abundant expression of the regulatory gene LIN28. Low let-7 levels resulted in up-regulation of oncogenes including MYCN, AURKB and LIN28 itself, the latter through a direct feedback mechanism. Targeting LIN28, or restoring let-7 levels, both led to effective inhibition of this pathway. In summary, paediatric malignant GCTs show biological differences from their adult counterparts at a genomic and protein-coding transcriptome level, whereas they both display very similar microRNA expression profiles. These similarities and differences may be exploited for diagnostic and/or therapeutic purposes.

Serum levels of mature microRNAs in DICER1-mutated pleuropulmonary blastoma.

DICER1 is a critical gene in the biogenesis of mature microRNAs, short non-coding RNAs that derive from either -3p or -5p precursor microRNA strands. Germline mutations of DICER1 are associated with a range of human malignancies, including pleuropulmonary blastoma (PPB). Additional somatic 'hotspot' mutations in the microRNA processing ribonuclease IIIb (RNase IIIb) domain of DICER1 are reported in cancer, and which affect microRNA biogenesis, resulting in a -3p mature microRNA strand bias. Here, in a germline (exon11 c.1806_1810insATTGA) DICER1-mutated PPB, we first confirmed the presence of an additional somatic RNase IIIb hotspot mutation (exon25 c.5425G>A [p.G1809R]) by conventional sequencing. Second, we investigated serum levels of mature microRNAs at the time of PPB diagnosis, and compared the findings with serum results from a comprehensive range of pediatric cancer patients and controls (n=52). We identified a panel of 45 microRNAs that were present at elevated levels in the serum at the time of PPB diagnosis, with a significant majority noted be derived from the -3p strand (P=0.013). In addition, we identified a subset of 10 serum microRNAs (namely miR-125a-3p, miR-125b-2-3p, miR-380-5p, miR-125b-1-3p, let-7f-2-3p, let-7a-3p, let-7b-3p, miR-708-3p, miR-138-1-3p and miR-532-3p) that were most abundant in the PPB case. Serum levels of two representative microRNAs, miR-125a-3p and miR-125b-2-3p, were not elevated in DICER1 germline-mutated relatives. In the PPB case, serum levels of miR-125a-3p and miR-125b-2-3p increased before chemotherapy, and then showed an early reduction following treatment. These microRNAs may offer future utility as serum biomarkers for screening patients with known germline DICER1 mutations for early detection of PPB, and for potential disease-monitoring in cases with confirmed PPB.

Bioactivity and biocompatibility of a chitosan-tobermorite composite membrane for guided tissue regeneration.

A polymer-mineral composite membrane of the mucopolysaccharide derivative, chitosan, and calcium silicate hydrate phase, tobermorite, was prepared by solvent casting and characterised by scanning electron microscopy (SEM) and Fourier Transform infrared spectroscopy (FTIR). The bioactivity and biocompatibility of the chitosan-tobermorite composite were evaluated in vitro with respect to its potential for use as a biodegradable guided tissue regeneration (GTR) membrane. The in vitro bioactivity of the composite was confirmed by the formation of crystalline substituted hydroxyapatite on the surface of the embedded tobermorite particles in simulated body fluid. The presence of the composite membrane was found to enhance the growth of MG63 human osteosarcoma cells by up to 30%. The findings of this initial study have indicated that this novel chitosan-tobermorite composite may be a suitable material for GTR applications.

A paucity of liver disease in Canadian Inuit with chronic hepatitis B virus, subgenotype B6 infection.

Clinical observations suggest that chronic hepatitis B virus (HBV) infections in the Canadian Inuit are less often associated with serious adverse outcomes than has been described in other HBV-infected patient populations. The aim of this study was to document the clinical and biochemical features, liver-related morbidity and all-cause mortality in Canadian Inuit with chronic HBV infections. Administrative databases were reviewed for individuals identified as hepatitis B surface antigen (HBsAg) positive during a 1983-85 seroepidemiological survey of viral hepatitis in Baffin Island, Canada. An equal number of age- and gender-matched HBsAg-negative individuals from the same communities served as controls. Baseline HBV viral loads, genotypes and specific mutations were compared in HBsAg-positive survivors and nonsurvivors. A subset of surviving HBsAg-positive carriers were reassessed 25-30 years following their initial diagnosis for evidence of advanced liver disease and changes to their serological/virological findings. One hundred and forty four HBsAg-positive individuals were identified. All were Canadian Inuit. The mean age at diagnosis was 38 ± 17 years and 69 (61%) were male. Median follow-up was 23 years (range: 2-28 years). Viral quantitation from stored sera could be performed in 70 infected individuals. The median viral load was 4.3 log 10 IU/ml (range: 2.3-8.8 log 10 IU/ml), and all were genotype B, subgenotype B6. Liver biochemistry, morbidity and all-cause mortality rates were similar in HBsAg-positive carriers and controls. Following multivariate analyses, only age at diagnosis predicted mortality in HBsAg carriers. In a subset of 30 HBsAg-positive survivors who underwent follow-up assessments, clinical, biochemical and radiological examinations of the liver were essentially normal. 23/30 (77%) remained HBsAg positive and 17/19 (90%) HBV-DNA positive. The genotype and prevalence of genomic mutations in this cohort remained largely unchanged, but quantifiable viral loads were significantly lower (P < 0.003). The results of this study suggest that chronic HBV infections in the Canadian Inuit are infrequently associated with serious adverse outcomes. Whether this finding reflects unique features of the host, presence or absence of external factors that influence the course of HBV and/or intrinsic properties of the HBV B6 subgenotype remains to be determined.

DICER1 hotspot mutations in non-epithelial gonadal tumours.

BACKGROUND: Non-epithelial gonadal tumours largely comprise sex cord-stromal tumours (SCSTs) and germ cell tumours (GCTs). Specific somatic mutations in DICER1, a microRNA maturation pathway gene, have been identified in these tumours. We conducted a study that aimed to confirm, refine and extend the previous observations. METHODS: We used Sanger sequencing to sequence the RNase IIIa and IIIb domains of DICER1 in 154 gonadal tumours from 135 females and 19 males, as well as 43 extra-gonadal GCTs from 26 females and 17 males. RESULTS: We identified heterozygous non-synonymous mutations in the RNase IIIb domain of DICER1 in 14/197 non-epithelial tumours (7.1%). Mutations were found in 9/28 SCSTs (32%), 5/118 gonadal GCTs (4.2%), 0/43 extra-gonadal GCTs and 0/8 miscellaneous tumours. The 14 mutations affected only five residues: E1705, D1709, E1788, D1810 and E1813. In all five patients where matched and constitutional DNA was available, the mutations were only somatic. There were no mutations found in the RNase IIIa domain. CONCLUSION: More than half (8/15) of Sertoli-Leydig cell tumours (SLCTs) harbour DICER1 mutations in the RNase IIIb domain, while mutations are rarely found in GCTs. Genetic alterations in SLCTs may aid in classification and provide new approaches to therapy.

Lack of correlation between predicted and actual off-target effects of short-interfering RNAs targeting the human papillomavirus type 16 E7 oncogene.

BACKGROUND: When designing therapeutic short-interfering RNAs (siRNAs), off-target effects (OTEs) are usually predicted by computational quantification of messenger RNAs (mRNAs) that contain matches to the siRNA seed sequence in their 3' UTRs. It is assumed that the higher the number of predicted transcriptional OTEs, the greater the size of the actual OTE signature and the more detrimental the phenotypic consequences in target-negative cells. METHODS: We tested this general assumption by investigating the OTEs of potential therapeutic siRNAs targeting the human papillomavirus (HPV) type-16 E7 oncogene. We studied HPV-negative squamous epithelial cells, from normal cervix (NCx) and skin (HaCaT), which would be vulnerable to 'bystander' OTEs following transfection in vivo. RESULTS: We observed no correlation between the number of computationally predicted OTEs and the actual number of seed-dependent OTEs (P=0.76). On average only 20.5% of actual transcriptional OTEs were seed-dependent (i.e., predicted). The unpredicted OTEs included stimulation of innate immune pathways, as well as indirect (downstream) effects of other OTEs, which affected important cancer-associated pathways. Although most significant OTEs observed were seen in both NCx and HaCaT cells, only 0-5.9% of differentially expressed genes overlapped between the two cell types. CONCLUSION: These data do not support the assumption that actual OTEs correlate well with predicted OTEs.

Recurring pulmonary hamartomas: cause for concern?

We report the case of a well-controlled female asthmatic who developed 'multiple pulmonary hamartomas' on three separate occasions over a period of 25 years that necessitated surgical resection. To our knowledge, this is the first report of recurrent hamartomas in a single individual necessitating multiple thoracotomies.

Diagnosis of bladder cancer by immunocytochemical detection of minichromosome maintenance protein-2 in cells retrieved from urine.

BACKGROUND: We tested the accuracy of immunocytochemistry (ICC) for minichromosome maintenance protein-2 (MCM-2) in diagnosing bladder cancer, using cells retrieved from urine. METHODS: Adequate samples were obtained from 497 patients, the majority presenting with gross haematuria (GH) or undergoing cystoscopic surveillance (CS) following previous bladder cancer. We performed an initial study of 313 patients, followed by a validation study of 184 patients. In all cases, presence/absence of bladder cancer was established by cystoscopy/biopsy. RESULTS: In the initial study, receiver operator characteristic analysis showed an area under the curve of 0.820 (P<0.0005) for the GH group and 0.821 (P<0.01) for the CS group. Optimal sensitivity/specificity were provided by threshold values of 50+ MCM-2-positive cells in GH samples and 200+ cells in CS samples, based on a minimum total cell number of 5000. Applying these thresholds to the validation data set gave 81.3% sensitivity, 76.0% specificity and 92.7% negative predictive value (NPV) in GH and 63.2% sensitivity, 89.9% specificity and 89.9% NPV in CS. Minichromosome maintenance protein-2 ICC provided clinically relevant improvements over urine cytology, with greater sensitivity in GH and greater specificity in CS (P=0.05). CONCLUSIONS: Minichromosome maintenance protein-2 ICC is a reproducible and accurate test that is suitable for both GH and CS patient groups.

An alternative method for predicting size of femoral component of Oxford partial knee replacement.

BACKGROUND: No direct intra-operative measurement to determine the ideal size of the femoral component of Oxford unicompartmental knee replacement (UKR) is currently present. The aim of this study is to assess the accuracy of patients' shoe size as a predictor of femoral component size. METHODS: A retrospective study was conducted to identify the correlation between patients' shoe size (British system) and the femoral component size. After excluding patients who died (n = 2) and patients in whom the implanted femoral component size was inaccurate (n = 13), the remaining cases (93 UKR in 88 patients) formed the study sample. Postoperative radiographs were reviewed to determine femoral component fit. RESULTS: We found positive correlation between shoe size and femoral component size. In females; a shoe size from 2.5 to 6 predicted a small femoral component and shoe size from 6.5 to 8.0 predicted a medium femoral component. In males, a shoe size from 6 to 9.5 predicted a medium femoral component and a shoe size from 10 to 13 predicted a large femoral component. This relation predicted the femoral component size accurately in 80% of cases. A subgroup analysis, after excluding patients who changed their shoe size during adulthood after foot surgery or pathology (n = 20), showed an accuracy rate of 81%. CONCLUSION: Shoe size is a simple method that predicts femoral component size more accurately than other methods currently used such as templating, tibial component size and height based on gender.

Lipoprotein lipase is frequently overexpressed or translocated in cervical squamous cell carcinoma and promotes invasiveness through the non-catalytic C terminus.

BACKGROUND: We studied the biological significance of genes involved in a novel t(8;12)(p21.3;p13.31) reciprocal translocation identified in cervical squamous cell carcinoma (SCC) cells. METHODS: The rearranged genes were identified by breakpoint mapping, long-range PCR and sequencing. We investigated gene expression in vivo using reverse-transcription PCR and tissue microarrays, and studied the phenotypic consequences of forced gene overexpression. RESULTS: The rearrangement involved lipoprotein lipase (LPL) and peroxisome biogenesis factor-5 (PEX5). Whereas LPL-PEX5 was expressed at low levels and contained a premature stop codon, PEX5-LPL was highly expressed and encoded a full-length chimeric protein (including the majority of the LPL coding region). Consistent with these findings, PEX5 was constitutively expressed in normal cervical squamous cells, whereas LPL expression was negligible. The LPL gene was rearranged in 1 out of 151 cervical SCCs, whereas wild-type LPL overexpression was common, being detected in 10 out of 28 tissue samples and 4 out of 10 cell lines. Forced overexpression of wild-type LPL and PEX5-LPL fusion transcripts resulted in increased invasiveness in cervical SCC cells, attributable to the C-terminal non-catalytic domain of LPL, which was retained in the fusion transcripts. CONCLUSION: This is the first demonstration of an expressed fusion gene in cervical SCC. Overexpressed wild-type or translocated LPL is a candidate for targeted therapy.

Methylator phenotype of malignant germ cell tumours in children identifies strong candidates for chemotherapy resistance.

BACKGROUND: Yolk sac tumours (YSTs) and germinomas are the two major pure histological subtypes of germ cell tumours. To date, the role of DNA methylation in the aetiology of this class of tumour has only been analysed in adult testicular forms and with respect to only a few genes. METHODS: A bank of paediatric tumours was analysed for global methylation of LINE-1 repeat elements and global methylation of regulatory elements using GoldenGate methylation arrays. RESULTS: Both germinomas and YSTs exhibited significant global hypomethylation of LINE-1 elements. However, in germinomas, methylation of gene regulatory regions differed little from control samples, whereas YSTs exhibited increased methylation at a large proportion of the loci tested, showing a 'methylator' phenotype, including silencing of genes associated with Caspase-8-dependent apoptosis. Furthermore, we found that the methylator phenotype of YSTs was coincident with higher levels of expression of the DNA methyltransferase, DNA (cytosine-5)-methyltransferase 3B, suggesting a mechanism underlying the phenotype. CONCLUSION: Epigenetic silencing of a large number of potential tumour suppressor genes in YSTs might explain why they exhibit a more aggressive natural history than germinomas and silencing of genes associated with Caspase-8-dependent cell death might explain the relative resistance of YSTs to conventional therapy.

Development of peptide aptamer microarrays for detection of HPV16 oncoproteins in cell extracts.

Protein microarrays represent an emerging technology that promises to facilitate high-throughput proteomics. The major goal of this technology is to employ peptides, full-length proteins, antibodies, and small molecules to simultaneously screen thousands of targets for potential protein-protein interactions or modifications of the proteome. This article describes the performance of a set of peptide aptamers specific for the human papillomavirus (HPV) type 16 oncoproteins E6 and E7 in a microarray format. E6 and E7 peptide aptamer microarrays were probed with fluorescence-labeled lysates generated from HPV-infected cervical keratinocytes expressing both E6 and E7 oncoproteins. Peptide aptamer microarrays are shown to detect low levels of E6 and E7 proteins. Peptide aptamers specific for cellular proteins included on these microarrays suggested that expression of CDK2, CDK4, and BCL-6 may be affected by HPV infection and genome integration. We conclude that peptide aptamer microarrays represent a promising tool for proteomics and may be of value in biological and clinical investigations of cervical carcinogenesis.

The use of cerclage wires as a suture passer in the repair of quadriceps tendon ruptures.

We describe a novel, simple and cost-effective method of passing sutures through the patella, without the need for expensive or specialised equipment.