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Biomedical subjects

G Kelly

Publications and source records attributed to G Kelly.

At least 19 recordsLinked to original sources

HLA-E and NKG2A Mediate Resistance to BCG Immunotherapy in Non-Muscle-Invasive Bladder Cancer.

Bacillus Calmette-Guérin (BCG) is the first-line therapy for high-grade non-muscle-invasive bladder cancer (NMIBC), yet many patients experience recurrence due to immune evasion. We identify HLA-E and NKG2A as mediators of adaptive resistance involving chronic activation of NK and T cells in BCG-unresponsive tumors. Prolonged IFN-γ exposure enhances HLA-E and PD-L1 expression on recurrent tumors, accompanied by the accumulation of NKG2A+ NK and CD8 T cells. HLA-Ehigh tumor cells preferentially cluster near CXCL12-rich stromal regions with dense effector cell presence, underscoring a spatially segregated tumor architecture. Although cytotoxic lymphocytes retain effector potential, their activity is restrained by HLA-E/NKG2A and PD-L1/PD-1 pathways located in their immediate neighborhood within the bladder tumor microenvironment. These data reveal a spatially organized immune escape program that limits anti-tumor immunity. Our findings support dually targeting NKG2A and PD-L1 checkpoint blockade as a rational, bladder-sparing strategy for patients with BCG-unresponsive NMIBC.

BCG-unresponsive↗

A positive role for PEA3 in HER2-mediated breast tumour progression.

Overexpression of HER2 is associated with an adverse prognosis in breast cancer. Despite this, the mechanism of its transcriptional regulation remains poorly understood. PEA3, a MAP kinase (MAPK)-activated member of the Ets transcription factor family has been implicated in the transcriptional regulation of HER2. The direction of its modulation remains controversial. We assessed relative levels of PEA3 expression and DNA binding in primary breast cultures derived from patient tumours (n=18) in the presence of an activated MAPK pathway using Western blotting and shift analysis. Expression of PEA3 in breast tumours from patients of known HER2 status (n=107) was examined by immunohistochemistry. In primary breast cancer cell cultures, growth factors induced interaction between PEA3 and its DNA response element. Upregulation of PEA3 expression in the presence of growth factors associated with HER2 positivity and axillary lymph node metastasis (P=0.034 and 0.049, respectively). PEA3 expression in breast cancer tissue associated with reduced disease-free survival (P<0.001), Grade III tumours (P<0.0001) and axillary lymph node metastasis (P=0.026). Co-expression of PEA3 and HER2 significantly associated with rate of recurrence compared to patients who expressed HER2 alone (P=0.0039). These data support a positive role for PEA3 in HER2-mediated oncogenesis in breast cancer.

Axilla↗

Otitis media and a neck lump--current diagnostic challenges for Paragonimus-like trematode infections.

A 29 year-old Nigerian studying in the UK presented with a neck lump and otitis media. Paragonimus-like trematode eggs were found in the neck lump aspirate. Morphologically these eggs resembled Paragonimus uterobilateralis or Achillurbainia congolensis. We favoured the diagnosis of achillurbainiasis over extrapulmonary paragonimiasis on the basis of clinical features and because we could not amplify DNA sequences using PCR primers specific for Paragonimus species. We discuss current diagnostic challenges for this rare parasitic infection.

Adult↗

Disease severity and genetic pathways in attenuated familial adenomatous polyposis vary greatly but depend on the site of the germline mutation.

BACKGROUND: Attenuated familial adenomatous polyposis (AFAP) is associated with germline mutations in the 5', 3', and exon 9 of the adenomatous polyposis coli (APC) gene. These mutations probably encode a limited amount of functional APC protein. METHODS AND RESULTS: We found that colonic polyp number varied greatly among AFAP patients but members of the same family tended to have more similar disease severity. 5' Mutants generally had more polyps than other patients. We analysed somatic APC mutations/loss of heterozygosity (LOH) in 235 tumours from 35 patients (16 families) with a variety of AFAP associated germline mutations. In common with two previous studies of individual kindreds, we found biallelic changes ("third hits") in some polyps. We found that the "third hit" probably initiated tumorigenesis. Somatic mutation spectra were similar in 5' and 3' mutant patients, often resembling classical FAP. In exon 9 mutants, in contrast, "third hits" were more common. Most "third hits" left three 20 amino acid repeats (20AARs) on the germline mutant APC allele, with LOH (or proximal somatic mutation) of the wild-type allele; but some polyps had loss of the germline mutant with mutation leaving one 20AAR on the wild-type allele. CONCLUSIONS: We propose that mutations, such as nt4661insA, that leave three 20AARs are preferentially selected in cis with some AFAP mutations because the residual protein function is near optimal for tumorigenesis. Not all AFAP polyps appear to need "three hits" however. AFAP is phenotypically and genetically heterogeneous. In addition to effects of different germline mutations, modifier genes may be acting on the AFAP phenotype, perhaps influencing the quantity of functional protein produced by the germline mutant allele.

Adenomatous Polyposis Coli↗

Transforming growth factor-beta (TGF-beta) in acute injuries of the spinal cord.

Injuries to the spinal cord may be associated with increased healing of fractures. This can be of benefit, but excessive bone growth can also cause considerable adverse effects. We evaluated two groups of patients with fractures of the spinal column, those with neurological compromise (n=10) and those without (n=15), and also a control group with an isolated fracture of a long bone (n=12). The level of transforming growth factor-beta (TGF-beta), was measured at five time points after injury (days 1, 5, 10, 42 and 84). The peak level of 142.79 ng/ml was found at day 84 in the neurology group (p<0.001 vs other time points). The other groups peaked at day 42 and had a decrease at day 84 after injury (p<or=0.001). Our findings suggest that TGF-beta may have a role in the increased bone turnover and attendant complications seen in patients with acute injuries to the spinal cord.

Acute Disease↗

How I do it? Excision of neck lumps under local anaesthesia.

Excisions of neck lesions under local anaesthesia are made technically more difficult by the injection of anaesthetic agent, causing the lesion to become less palpable. Pen marking the lesion can circumvent the problem; however, this marking often disappears after skin preparation. We describe a technique which abolishes this problem.

Anesthesia, Local↗

Inverse relationship between ER-beta and SRC-1 predicts outcome in endocrine-resistant breast cancer.

The oestrogen receptor (ER) interacts with coactivator proteins to modulate genes central to breast tumour progression. Oestrogen receptor is encoded for by two genes, ER-alpha and ER-beta. Although ER-alpha has been well characterized, the role of ER-beta as a prognostic indicator remains unresolved. To determine isoform-specific expression of ER and coexpression with activator proteins, we examined the expression and localisation of ER-alpha, ER-beta and the coactivator protein steroid receptor coactivator 1 (SRC-1) by immunohistochemistry and immunofluorescence in a cohort of human breast cancer patients (n=150). Relative levels of SRC-1 in primary breast cultures derived from patient tumours in the presence of beta-oestradiol and tamoxifen was assessed using Western blotting (n=14). Oestrogen receptor-beta protein expression was associated with disease-free survival (DFS) and inversely associated with the expression of HER2 (P=0.0008 and P<0.0001, respectively), whereas SRC-1 was negatively associated with DFS and positively correlated with HER2 (P<0.0001 and P<0.0001, respectively). Steroid receptor coactivator 1 protein expression was regulated in response to beta-oestradiol or tamoxifen in 57% of the primary tumour cell cultures. Protein expression of ER-beta and SRC-1 was inversely associated (P=0.0001). The association of ER-beta protein expression with increased DFS and its inverse relationship with SRC-1 suggests a role for these proteins in predicting outcome in breast cancer.

Adult↗

Estimation of infection prevalence from correlated binomial samples.

Infection prevalence in a population often is estimated from grouped binary data expressed as proportions. The groups can be families, herds, flocks, farms, etc. The observed number of cases generally is assumed to have a Binomial distribution and the estimate of prevalence is then the sample proportion of cases. However, the individual binary observations might not be independent--leading to overdispersion. The goal of this paper was to demonstrate random-effects models for the estimation of infection prevalence from data which are correlated and in particular, to illustrate a nonparametric random-effects model for this purpose. The nonparametric approach is a relatively recent addition to the random-effects class of models and does not appear to have been discussed previously in the veterinary epidemiology literature. The assumptions for a logistic-regression model with a nonparametric random effect were outlined. In a demonstration of the method on data relating to Salmonella infection in Irish pig herds, the nonparametric method resulted in the classification of herds into a small number of distinct prevalence groups (i.e. low, medium and high prevalence) and also estimated the relative frequency of each prevalence category in the population. We compared the estimates from a logistic model with a nonparametric distribution for the random effects with four alternative models: a logistic-regression model with no random effects, a marginal model using a generalised estimating equation (GEE) and two methods of fitting a Normally distributed random effect (the GLIMMIX macro and the NLMIXED procedure both in SAS). Parameter estimates from random-effects models are not readily interpretable in terms of prevalences. Therefore, we outlined two methods for calculating population-averaged estimates of prevalence from random-effects models: one using numerical integration and the other using Monte Carlo simulation.

Animals↗

Consent for tonsillectomy.

Rulings in recent negligence cases reveal a shift towards what the 'reasonable patient' would expect in deciding the risks doctors must disclose to patients. This survey aimed to investigate whether the 'reasonable patient' and 'responsible body of medical opinion' agree about which risks should be discussed regarding tonsillectomy. Using questionnaires, surgeons were asked which of the 10 complications they routinely discussed and patients were asked how seriously they regarded these complications. The results were compared with the Test of Proportions. Most surgeons routinely mentioned otalgia, odynophagia, throat infection and re-operation. Most patients regarded potentially fatal bleeding, pneumonia and blood transfusion as very serious but only the minority of surgeons mentioned these (P < 0.001). When obtaining consent for tonsillectomy, surgeons do not routinely mention all the risks that the 'reasonable patient' would expect. The 'reasonable patient' would expect that re-operation, transfusion, pneumonia and fatal blood loss are discussed.

Blood Transfusion↗

Expression of SRC-1, AIB1, and PEA3 in HER2 mediated endocrine resistant breast cancer; a predictive role for SRC-1.

BACKGROUND: In human breast cancer, the growth factor receptor HER2 is associated with disease progression and resistance to endocrine treatment. Growth factor induced mitogen activated protein kinase activity can phosphorylate not only the oestrogen receptor, but also its coactivator proteins AIB1 and SRC-1. AIM: To determine whether insensitivity to endocrine treatment in HER2 positive patients is associated with enhanced expression of coactivator proteins, expression of the HER2 transcriptional regulator, PEA3, and coregulatory proteins, AIB1 and SRC-1, was assessed in a cohort of patients with breast cancer of known HER2 status. METHODS: PEA3, AIB1, and SRC-1 protein expression in 70 primary breast tumours of known HER2 status (HER2 positive, n = 35) and six reduction mammoplasties was assessed using immunohistochemistry. Colocalisation of PEA3 with AIB1 and SRC-1 was determined using immunofluorescence. Expression of PEA3, AIB1, and SRC-1 was correlated with clinicopathological parameters. RESULTS: In primary breast tumours expression of PEA3, AIB1, and SRC-1 was associated with HER2 status (p = 0.0486, p = 0.0444, and p = 0.0012, respectively). In the HER2 positive population, PEA3 expression was associated with SRC-1 (p = 0.0354), and both PEA3 and SRC-1 were significantly associated with recurrence on univariate analysis (p = 0.0345; p<0.0001). On multivariate analysis, SRC-1 was significantly associated with disease recurrence in HER2 positive patients (p = 0.0066). CONCLUSION: Patients with high expression of HER2 in combination with SRC-1 have a greater probability of recurrence on endocrine treatment compared with those who are HER2 positive but SRC-1 negative. SRC-1 may be an important predictive indicator and therapeutic target in breast cancer.

Adult↗

Prevalence and number of Salmonella in irish retail pork sausages.

A national Salmonella control program in the pork industry was enacted in Ireland in August 2002. This study was undertaken as part of a larger project investigating the role of pork as a source of human salmonellosis in Ireland. The objective of this survey was to assess the prevalence of Salmonella in Irish pork sausage at retail level. Samples, comprising branded prepacked sausages and loose sausages from supermarket meat counters and butcher shops, were collected from selected retail sites in four cities from October to December 2001 and from June to August 2002. A three-tube most-probable-number method was used to enumerate Salmonella in a selected number of samples that were positive by enrichment. Salmonella serotypes were detected in 4.4 and 1.7% of samples at each of the respective sampling periods, a level similar to those reported in other U.S. and U.K. studies. Isolates were characterized by serotype, phage type, and antimicrobial susceptibility. Eighteen (70%) were resistant to at least one antimicrobial, and 15 (58%) were resistant to four or more antimicrobials. Most of the isolates exhibited resistance to tetracycline. Five different phage types were detected. DT104 was the predominant phage type among Salmonella Typhimurium isolates. This study revealed that multidrug-resistant salmonellae are present in a proportion of Irish sausages and that further risk analysis work is necessary in order to quantify the risk posed to public health.

Animals↗

Salmonella serotypes present on a sample of Irish pig farms.

A survey of the prevalence of Salmonella species infection was conducted on 59 Irish farrow-to-finish pig herds. Faecal samples were collected from the pens of first-stage weaners (growing pigs approximately three to 10 weeks of age), second-stage weaners (approximately 10 to 17 weeks of age) and fatteners, and from the dry sow and farrowing sow houses. The prevalence of infection was estimated to within 5 per cent with a 95 per cent confidence interval. Thirty of the 59 herds were infected, 12 with Salmonella Typhimurium only, eight with Salmonella Derby only and seven with both S Typhimurium and S Derby; serotypes London, Livingstone and Infantis were each isolated from a single herd. Farms in Ireland are assigned to one of three infection categories on the basis of the antibody levels in samples of meat juice taken at slaughter. When a herd was classified as either positive or negative on the basis of the isolation of Salmonella from at least one faecal sample there was no association between the herd's category as determined by meat juice serology and the probability of the isolation of Salmonella from the faecal samples. However, there were differences in prevalence between pigs at different stages of production in herds of different categories. Farrowing sow houses in moderately infected (category 2) herds had significantly lower infection rates (P < or = 0.05) than other herd categories and other stages of production. Pigs from first-stage weaner pens in slightly infected (category 1) herds were more likely to be infected with Salmonella than pigs at any other stage of production or category of herd.

Animals↗

Idiopathic mucosal lesions of the arytenoid cartilages of 21 Thoroughbred yearlings: 1997-2001.

REASONS FOR PERFORMING STUDY: Mucosal ulcers and, occasionally, small granulomas on the axial surface of one or both arytenoid cartilages have been found in TB yearlings presented for post sale endoscopic examination. OBJECTIVES: To determine the incidence, endoscopic characteristics and outcome of a group of Thoroughbred yearlings affected with mucosal ulcers and granulomas of the arytenoid cartilage. HYPOTHESIS: The incidence of mucosal ulceration of the arytenoid cartilages of yearling Thoroughbreds is relatively high compared to other upper airway abnormalities; and that the majority of mucosal ulcers heal uneventfully, although a small percentage may progress to a granuloma and, less commonly, to arytenoid chondropathy. METHODS: The findings of post sale, upper airway endoscopic examinations of 3312 Thoroughbred yearlings, during a 5 year period, were reviewed, including those abnormalities listed in the conditions of sale and others not listed but considered likely to cause airway obstruction. Information obtained from the medical record of horses that had mucosal ulceration or granuloma of the arytenoid cartilage included the location and size of the lesion(s), sex of the affected horse and the presence and nature of other concurrent abnormalities of the upper portion of the respiratory tract. Additional information included treatment and results of follow-up, endoscopic examination by the authors or attending veterinarian. RESULTS: Mucosal lesions were seen in 0.63% of yearlings evaluated, which represented the most common, documented condition of the upper portion of the respiratory tract. The mucosal ulcers of 15 of 19 horses were considered to have healed without complication during follow-up examination; one of the 19 horses was lost to follow-up. Two horses affected with bilateral, arytenoid mucosal ulceration developed a granuloma at each site of ulceration. One horse developed a granuloma at a site of ulceration and, subsequently, arytenoid chondropathy. CONCLUSIONS: Arytenoid mucosal ulceration in sales yearlings was a relatively commonly encountered abnormality and a small percentage progressed to granuloma or chondropathy. POTENTIAL RELEVANCE: The mucosa of the arytenoid cartilage, particularly at the rostral margin of the vocal process, should be examined carefully during endoscopic examination of the upper portion of the respiratory tract of Thoroughbred yearlings presented for sale. Because a small percentage of mucosal ulcers may progress to granuloma or, less commonly, chondropathy, identification of mucosal ulcers of the arytenoid cartilage seen during post sale, endoscopic examination warrants notification to the purchaser and sales company. Medical therapy of affected horses should be considered and follow-up endoscopic examination performed to determine if the lesion has healed.

Animals↗

Myringoplasty: does the size of the perforation matter?

Does the size of a tympanic membrane defect influence the success rate in myringoplasty? Many studies which have suggested that size does not affect results have included only small numbers of patients. After a power analysis to identify the size of groups needed to detect a clinically significant difference between the success rate of small and large perforations, a retrospective study was carried out to answer the question. A retrospective study of 423 myringoplasty-only operations showed that size does influence success; the success rate for small perforations was 74.1% compared with 56.0% for large perforations (P = 0.0003). Size also influences the change in four-frequency air conduction thresholds (small perforations +7.2 dB, large perforations +10.2 dB; t = 2.08, P = 0.039) in successful myringoplasties, a positive value indicating an improvement in hearing. This change is unlikely to be clinically significant. Patients should be made aware of these facts when informed consent is being obtained.

Audiometry↗

Survival of Escherichia coli O157:H7 in farm water: its role as a vector in the transmission of the organism within herds.

AIMS: The study aimed to investigate the survival characteristics of Escherichia coli O157:H7 in farm water (FW), and in sterile distilled municipal water (SDW), stored outdoors under field conditions, with or without the addition of faeces (1% w/v), in a farmyard shed and the laboratory at 15 degrees C. METHODS AND RESULTS: Water samples were inoculated with E. coli O157:H7 at 10(3) and 10(6) ml(-1), and sampled over a 31-day period. In FW stored outdoors in a field, E. coli O157:H7 survived for 14 days at temperatures <15 degrees C, at both inoculation levels, while in the laboratory at 15 degrees C, the organism was still detectable at low levels (<1 log10 cfu ml(-1)) after 31 days. The addition of bovine faeces to water outdoors (1% w/v) resulted in survival for 24 days. In SDW inoculated at 10(6) ml(-1) and stored in the laboratory (15 degrees C), only a 2.5 log reduction was observed after 31 days, while the organism could not be detected after 17 days in the field. Preliminary screening of water samples stored outdoors isolated a bacterium which exhibited antimicrobial activity towards E. coli O157:H7. CONCLUSIONS: The survival of E. coli O157:H7 observed in this study illustrates the potential of farm water to act as a vehicle in the transfer of the organism across a herd. SIGNIFICANCE AND IMPACT OF THE STUDY: The difficulty in extrapolating results from controlled laboratory situations to on-farm conditions is also highlighted in this study.

Agriculture↗

An investigation of mucus/polymer rheological synergism using synthesised and characterised poly(acrylic acid)s.

A range of poly(acrylic acid)s with different average degrees of polymerisation and cross-linking densities were synthesised using a solution polymerisation process. The rheological characteristics of aqueous dispersions of these materials and those of mixtures with homogenised pigs gastric mucus were investigated using dynamic oscillatory rheology, and compared to the known mucoadhesive Carbopol 934P. From the storage moduli, the rheological synergy and relative rheological synergy were calculated, and the effects of concentration and pH on this considered. Generally, the larger the molecular weight (and degree of cross-linking), the greater the rheological synergy, with Carbopol 934P giving the most pronounced effect. Rheological synergy was seen to be concentration-dependent, and a maximum concentration to produce an optimum effect was evident. Acid pHs were seen to favour synergy, although in marked contrast to previous literature reports, the optimum mucus-polymer interaction was not observed at the half ionised value (pH = pKa) but at pH regimes that were unique to each polymer type. This could be influenced by the structural constrains imposed on potential hydrogen bonded interactions. It was concluded that synthesising poly(acrylic acid)s with better defined physicochemical properties than commercially available polymers will advance the study of the phenomenon of rheological synergy.

Acrylic Resins↗

The Bowman-Birk inhibitor reactive site loop sequence represents an independent structural beta-hairpin motif.

We have determined the NMR structure in aqueous solution of a disulphide-cyclised 11-residue peptide that forms a stable beta-hairpin, incorporating a type VIb beta-turn. The structure is found to be extremely well ordered for a short peptide, with the 30 lowest energy simulated annealing structures having an average pairwise r.m.s. deviation of only 0.36 A over the backbone. All but three side-chains adopt distinct conformations, allowing a detailed analysis of their involvement in cross-strand interactions. The peptide sequence analysed originates from a previously reported study, which identified potent inhibitors of human leukocyte elastase from screening a combinatorial peptide library based on the short protein beta-sheet segment that forms the reactive site loop of Bowman-Birk inhibitors. A detailed comparison of the peptide's solution structure with the corresponding region in the whole protein structure reveals a very good correspondence not only for the backbone (r.m.s. deviation approximately 0.7 A) but also for the side-chains. This isolated beta-hairpin retains the biologically active "canonical conformation" typical of small serine proteinase inhibitor proteins, which explains why it retains inhibitory activity. Since the structural integrity is sequence-inherent and does not depend upon the presence of the remaining protein, this beta-hairpin represents an independent structural motif and so provides a useful model of this type of protein architecture and its relation to biological function. The relationship between the conformation of this beta-hairpin and its biological activity is discussed.

Amino Acid Motifs↗