Intestinal perforation in the premature infant.

OBJECTIVE: To compare demographic data, prenatal and postnatal characteristics, laboratory data, and outcomes in a cohort of premature infants with spontaneous ileal perforation (SIP), surgical necrotizing enterocolitis (sNEC) and matched controls. METHODS: A retrospective case-control study of infants with intestinal perforation with a birth weight (BW) less than 2,000 grams and gestational age (GA) less than 34 weeks and infants without perforation matched for BW (±150 grams) and GA (±1week). RESULTS: 130 premature infants were included, 30 infants with SIP, 35 infants with sNEC and 65 control infants. The median age of onset was 5 days postnatal age in SIP versus 25 days in sNEC (p < 0.001) and the peak onset was at 26 weeks corrected GA for SIP and 30 weeks corrected GA for sNEC. Infants with perforation had significantly higher rates of mortality (p < 0.001) and common morbidities associated with prematurity. Administration of corticosteroids and indomethacin did not differ among groups. SIP was more common among infants born to young mothers (p = 0.04) and less common in infants receiving caffeine (p = 0.02). sNEC was less common among infants receiving early red cell transfusion (p = 0.01). Perforation and sNEC trended towards less common in infants receiving inhaled nitric oxide. CONCLUSION: SIP and sNEC are distinct clinical entities. Potential protective effects of caffeine, inhaled nitric oxide, and early transfusion should be further studied.

Buccal administration of human colostrum: impact on the oral microbiota of premature infants.

OBJECTIVE: To determine whether the administration of mother's colostrum into the buccal pouch in the first days of life alters the oral microbiota compared with control infants. STUDY DESIGN: In this pilot study, 12 very low birth weight (VLBW) infants were randomly assigned to receive either colostrum from their mothers directly into the buccal pouch every 2 h for 46 h or standard care. We analyzed the oral microbiota at initiation and 48 and 96 h later using next-generation sequencing. RESULT: The oral microbiota changed markedly over the 96 h period in all babies. Patterns of colonization differed between groups with Planococcaceae, the dominant family at 48 and 96 h in the colostrum group, and Moraxellaceae and Staphylococcaceae, the dominant families at 48 and 96 h, respectively, in the control group. CONCLUSION: Buccal administration of mother's colostrum to VLBW infants influenced the colonization of the oral cavity with differences persisting 48 h after completion of the intervention.

Probiotic administration in congenital heart disease: a pilot study.

OBJECTIVE: To investigate the impact of probiotic Bifidobacterium longum ssp. infantis on the fecal microbiota and plasma cytokines in neonates with congenital heart disease. STUDY DESIGN: Sixteen infants with congenital heart disease were randomly assigned to receive either B. infantis (4.2 × 10(9) colony-forming units two times daily) or placebo for 8 weeks. Stool specimens from enrolled infants and from six term infants without heart disease were analyzed for microbial composition. Plasma cytokines were analyzed weekly in the infants with heart disease. RESULTS: Healthy control infants had increased total bacteria, total Bacteroidetes and total bifidobacteria compared to the infants with heart disease, but there were no significant differences between the placebo and probiotic groups. Plasma interleukin (IL)10, interferon (IFN)γ and IL1ß levels were transiently higher in the probiotic group. CONCLUSION: Congenital heart disease in infants is associated with dysbiosis. Probiotic B. infantis did not significantly alter the fecal microbiota. Alterations in plasma cytokines were found to be inconsistent.

Androgen receptor phosphorylation at serine 515 by Cdk1 predicts biochemical relapse in prostate cancer patients.

BACKGROUND: Prostate cancer cell growth is dependent upon androgen receptor (AR) activation, which is regulated by specific kinases. The aim of the current study is to establish if AR phosphorylation by Cdk1 or ERK1/2 is of prognostic significance. METHODS: Scansite 2.0 was utilised to predict which AR sites are phosphorylated by Cdk1 and ERK1/2. Immunohistochemistry for these sites was then performed on 90 hormone-naive prostate cancer specimens. The interaction between Cdk1/ERK1/2 and AR phosphorylation was investigated in vitro using LNCaP cells. RESULTS: Phosphorylation of AR at serine 515 (pAR(S515)) and PSA at diagnosis were independently associated with decreased time to biochemical relapse. Cdk1 and pCdk1(161), but not ERK1/2, correlated with pAR(S515). High expression of pAR(S515) in patients with a PSA at diagnosis of ≤20 ng ml(-1) was associated with shorter time to biochemical relapse (P=0.019). This translated into a reduction in disease-specific survival (10-year survival, 38.1% vs 100%, P<0.001). In vitro studies demonstrated that treatment with Roscovitine (a Cdk inhibitor) caused a reduction in pCdk1(161) expression, pAR(S515)expression and cellular proliferation. CONCLUSION: In prostate cancer patients with PSA at diagnosis of ≤20 ng ml(-1), phosphorylation of AR at serine 515 by Cdk1 may be an independent prognostic marker.

The accuracy of magnetic resonance imaging in radical prostatectomy.

AIMS: The aim of this study was to examine the accuracy of standard magnetic resonance imaging (MRI) in the localised staging of prostate cancer in those who had undergone radical prostatectomy. PATIENTS AND METHODS: The cohort consisted of 110 patients who had undergone MRI for staging of prostate cancer and subsequently underwent radical prostatectomy. T stage was analysed both on MRI and from the specimen following radical surgery. RESULTS: Of the patients 57% of patients had their disease up-staged following radical surgery from preoperative MRI findings. Of those patients who had their disease up-staged following surgery, nearly 50% of patients had gone from organ confined disease at time of MRI to extra-prostatic involvement from the surgical specimen. CONCLUSION: We have reported that MRI has a wide range of accuracy. Given developments in MRI technologies further work should be pursued to help in the staging of this disease for which decision to treat is difficult.

NFκB signalling is upregulated in a subset of castrate-resistant prostate cancer patients and correlates with disease progression.

BACKGROUND: Cell line models suggest that activation of NFκB is associated with progression of prostate cancer. This pathway may be a therapeutic target if these observations translate to clinical specimens. METHODS: Immunohistochemistry measured NFκBp65 (p65), NFκBp65 nuclear localisation signal (NLS), NFκBp65 phosphorylated at ser 276 (p65(ser276)), NFκBp65 phosphorylated at ser 536 (p65(ser536)), IκBα phosphorylated at ser 32/36 (pIκBα(ser32/36)) and MMP-9 protein expression in 61 matched hormone naive prostate cancer (HNPC) and castrate-resistant prostate cancer (CRPC) tumours. Animal and cell models were used to investigate the role of NFκB inhibition in prostate carcinogenesis. RESULTS: In HNPC tumours, NLS expression significantly associated with a shorter time to disease recurrence and disease-specific death. In CRPC tumours p65, pIκBα(ser32/36) and MMP-9 expression significantly associated with shorter time to death from disease recurrence and shorter disease-specific death. MMP-9 and pIκBα(ser32/36) expression significantly associated with metastases at recurrence and were independent of Gleason sum and prostate-specific antigen at recurrence. Expression of phosphorylated Akt was associated with increased p65 activation in mouse models and inhibition of NFκB in LNCaP cells significantly reduced cellular proliferation and induced apoptosis. CONCLUSION: These results provide further evidence that the NFκB pathway could be exploited as a target for CRPC.

Hyperleukocytosis in a premature infant with intrauterine herpes simplex encephalitis.

Herpes encephalitis is a rare but devastating infection in premature infants. We report a 29 week gestation infant with severe intrauterine cutaneous and central nervous system herpes accompanied by hyperleukocytosis. Leukemoid reactions are not uncommon in this population, but the association of herpes encephalitis and a leukemoid reaction or hyperleukocytosis has not been reported previously.

The interrelationships between Src, Cav-1 and RhoGD12 in transitional cell carcinoma of the bladder.

BACKGROUND: The aim of this current study was to assess the expression and activity of Src family kinases, focal adhesion kinase (FAK), caveolin (Cav-1) and RhoGD12 in bladder cancer. METHODS: Fifty-eight patients with a new diagnosis of bladder cancer undergoing transurethral resection were included. Immunohistochemical staining was utilised to assess expression of c-Src, dephosphorylated (SrcY(530)), phosphorylated Src (Y(419)), phosphorylated FAK (FAK Y(861)), Cav-1 and RhoGD12. Expression was assessed using the weighted histoscore method. RESULTS: High expression of dephosphorylated Y(527), phosphorylated Y(416) and phosphorylated FAK Y(861) in the membrane were associated with increased cancer-specific survival (P=0. 01, P=0.001, P=0.008, respectively) and expression of Y(416) in the membrane was an independent factor on multivariate analysis when combined with known clinical parameters (P=0.008, HR 0.288, 95% CI 0.11-0.72). CONCLUSION: These results demonstrate that in contrast to other solid tumours, activation of the Src family members and downstream signalling proteins are associated with a good prognosis in transitional cell carcinoma of the bladder, and activated Src has a positive relationship with RhoGD12.

Heregulin expression and prognosis in prostate adenocarcinoma.

BACKGROUND: Heregulins (HRG) are a family of growth factors acting as ligands to HER3, HER4 and HER family signalling. HRG has a concentration-dependent differential growth effect--low levels mitogenic, high levels inhibitory. HRG differentially affects hormone-naïve (inhibitory) and castrate-resistant (proliferative) prostate adenocarcinoma (CaP) cell lines. We postulate that differential HRG expression in CaP will be associated with alteration in tumour growth, development and prognosis. PATIENTS AND METHODS: HRG expression was assessed in 2 cohorts: cohort 1 of 45 patients with paired hormone-naïve and castrate-resistant samples, and cohort 2 of 357 hormone-naïve samples. Correlations between HRG expression and biochemical relapse and survival were determined. RESULTS: In cohort 1, hormone-naïve samples' high membranous HRG expression was associated with increased time to relapse (p = 0.036), time to death from relapse (p = 0.002) and overall survival (p = 0.001). Membrane HRG fell significantly in post-relapse specimens. In cohort 2, high membranous HRG was associated with increased time to relapse (p = 0.004) and overall survival (p = 0.044) in patients treated with castration therapy but only with overall survival (p = 0.002) in the full cohort. CONCLUSION: High HRG expression is associated with improved prognosis in hormone-naïve CaP and a fall in expression occurs at castration escape indicating a protective role against castrate resistance.

Upregulation of MAPK pathway is associated with survival in castrate-resistant prostate cancer.

BACKGROUND: Recent evidence has implicated the MAP kinase (MAPK) pathway with the development of castrate-resistant prostate cancer (CRPC). We have previously reported gene amplification of critical members of this pathway with the development of castrate-resistant disease. In addition, we have shown that rising Raf-1 expression, with the development of CRPC, influences time to biochemical relapse. We therefore sought to further analyse the role of both Raf-1 and its downstream target MAPK in the molecular pathogenesis of CRPC. METHODS: Protein expression of Raf-1 and MAPK, including their activation status, was analysed using immunohistochemistry in a database of 65 paired tumour specimens obtained before and after the development of CRPC and correlated with other members of the pathway. RESULTS: Patients whose nuclear expression of MAPK rose with the development of CRPC had a significantly shorter median time to death following biochemical relapse (1.40 vs 3.00 years, P=0.0255) as well as reduced disease-specific survival when compared with those whose expression fell or remained unchanged (1.16 vs 2.62 years, P=0.0005). Significant correlations were observed between protein expression of Raf-1 and MAPK with the type 1 receptor tyrosine kinases, Her2 and epidermal growth factor receptor, as well as the transcription factor AP-1 in CRPC tumours. CONCLUSION: We conclude that the Her2/Raf-1/MAPK/AP-1 axis may promote the development of CRPC, leading to early relapse, and reduced disease-specific survival. In addition, members of the pathway may act as novel therapeutic and/or diagnostic targets for prostate cancer.

Presence of tumoural C-reactive protein correlates with progressive prostate cancer.

C-reactive protein (CRP) is an acute phase protein implicated in the progression of cancer. A positive correlation between tumour stage and plasma CRP levels was demonstrated in prostate cancer, indicating a relationship between raised CRP levels and more aggressive disease, suggesting a role for inflammatory response in tumour progression. Aim of this study was to assess the tumoural presence and cellular location of CRP and establish if these are linked to clinicopathological features of the cohort and patient survival. Tissue microarray technology was employed to analyse 50 matched pairs of hormone sensitive and refractory prostate cancers. Immunohistochemistry was performed using antibody to CRP. CRP was assessed using the weighted histoscore method. CRP presence was observed in the cytoplasm and nucleus of selected tumours. Cytoplasmic CRP correlated positively with metastases at diagnosis (P=0.039), whereas nuclear CRP presence correlated with metastases at relapse (P=0.006). A trend towards an increase in cytoplasmic and nuclear CRP presence from hormone sensitive to hormone refractory tumours was noticed. No significant association between tumoural CRP presence, time to biochemical relapse or disease-specific survival was observed. Tumoural CRP is likely to have a role in progression of prostate cancer, as it is associated with increased presence of metastases at the time of diagnosis and time of relapse. A larger powered study is necessary to establish if CRP presence is associated with disease-specific survival.

Cost, causes and rates of rehospitalization of preterm infants.

OBJECTIVE: To examine hospital readmissions for premature infants during the first year of life. STUDY DESIGN: The California maternal and newborn/infant hospital discharge records were examined for subsequent readmission during the first year of life for all newborns from 1992 to 2000. Discharge diagnoses, hospital days, demographic data and hospital charges for infants born preterm (<36 weeks gestation) were identified and evaluated. RESULT: About 15% of preterm infants required at least one rehospitalization within the first year of life (average cost per readmission 8,468 dollars, average annual cost in excess of 41 million dollars). Infants with gestational age <25 weeks had the highest rate of readmission (31%) and longest average length of stay (12 hospital days). The largest cohort, infants born at 35 weeks gestation, had the highest total cost of readmission (92.9 million dollars). The most common cause of rehospitalization was acute respiratory disease. There was no decrease in the number or cost of readmissions of premature infants for respiratory syncytial virus infections following the introduction of palivizumab in 1998. CONCLUSION: After initial discharge, premature infants continue to have significant in-patient health-care needs and costs.

The relationship between the systemic inflammatory response, tumour proliferative activity, T-lymphocytic infiltration and COX-2 expression and survival in patients with transitional cell carcinoma of the urinary bladder.

The relationship between the systemic inflammatory response, tumour proliferative activity, T-lymphocytic infiltration, and COX-2 expression and survival was examined in patients with transitional cell carcinoma of the urinary bladder (n=103). Sixty-one patients had superficial disease and 42 patients had invasive disease. Cancer-specific survival was shorter in those patients with invasive compared with superficial bladder cancer (P<0.001). On univariate analysis, stratified by stage, increased Ki-67 labelling index (P<0.05), increased COX-2 expression (P<0.05), C-reactive protein (P<0.05) and adjuvant therapy (P<0.01) were associated with poorer cancer-specific survival. On multivariate analysis of these significant factors, stratified by stage, only C-reactive protein (HR 2.89, 95% CI 1.42-5.91, P=0.004) and adjuvant therapy (HR 0.29, 95% CI 0.14-0.62, P=0.001) were independently associated with poorer cancer-specific survival. These results would suggest that tumour-based factors such as grade, COX-2 expression or T-lymphocytic infiltration are subordinate to systemic factors such as C-reactive protein in determining survival in patients with transitional cell carcinoma of the urinary bladder.

The relationship between the systemic inflammatory response and survival in patients with transitional cell carcinoma of the urinary bladder.

The relationship between tumour stage, grade, elevated C-reactive protein concentration (<10/>10 mg l(-1)), adjuvant therapy and survival was examined in patients with biopsy proven bladder cancer (n=105). On multivariate analysis stage (HR 3.37, 95% CI 1.37-8.29, P=0.008), grade (HR 2.01, 95% CI 1.14-3.57, P=0.017) and preoperative C-reactive protein (HR 3.31, 95% CI 1.09-10.09, P=0.035) were independently associated with cancer-specific survival.

Raf-1 expression may influence progression to androgen insensitive prostate cancer.

BACKGROUND: Recent evidence has implicated the MAP kinase pathway with the development of androgen insensitive prostate cancer (AIPC). We have previously reported gene amplification of critical members of this pathway with the development of androgen insensitive disease. METHODS: Protein expression of Raf-1 was analyzed using immunohistochemistry (IHC) in a database of 65 paired tumor specimens obtained before and after the development of AIPC and correlated with other members of the pathway. RESULTS: Patients whose Raf-1 expression rose with development of AIPC had a significantly shorter median time to biochemical relapse compared to those whose expression fell or remained unchanged (1.16 vs. 2.62 years, P = 0.0005). In AIPC tumors, expression of Raf-1 correlated significantly with expression of HER2 and with expression of c-fos. CONCLUSIONS: We conclude that the HER2/Raf-1/AP-1 axis may promote the development of AIPC, leading to early relapse. Members of the pathway may act as novel therapeutic targets for patients.

The relationship between interleukin-6 and C-reactive protein in patients with benign and malignant prostate disease.

The relationship between interleukin-6 and C-reactive protein was evaluated in patients with benign (n=59) and malignant (n=86) prostate disease. The correlation coefficients for patients with benign prostatic disease and prostate cancer were rs=0.632, P<0.001 and rs=0.663, P<0.001, respectively. These results indicate that the relationship between interleukin-6 and C-reactive protein is similar in patients with benign and malignant prostate disease.

The relationship between T-lymphocyte subset infiltration and survival in patients with prostate cancer.

The relationship between tumour stage, T-lymphocyte subset infiltration and survival was examined in patients with prostate cancer (n=80). On multivariate analysis PSA (HR 2.47, 95% CI 1.27-4.83, P=0.008) and CD4+ T-lymphocyte count (HR 2.29, 95% CI 1.25-4.22, P=0.008) had independent significance. Increased CD4+ T-lymphocyte infiltration within the tumour was stage independent and associated with poor outcome in patients with prostate cancer.

Comparison of total, complexed and free prostate-specific antigens and their ratios in the detection of prostate cancer in a non-screened population.

BACKGROUND: The role of prostate-specific antigen (PSA) isoforms in the detection of prostate cancer in a non-screened population in the UK remains to be determined. METHODS: Consecutive patients undergoing diagnostic transrectal ultrasound-guided biopsy of the prostate were studied. Prior to biopsy, a blood sample was obtained and total, complexed and free PSA concentrations measured. RESULTS: Of the 171 patients included in the study, 103 were found to have prostate cancer. There were significant differences in total and complexed PSA concentrations and in the ratio of free-to-total PSA (all P <0.001) between patients with prostate cancer and those with benign disease. Receiver operating characteristics (ROC) curve analysis showed that the corresponding areas under the curves were similar. Restricting the analysis to the 77 patients who had total PSA concentrations between 2 and 10 micro g/L, ROC curve analysis showed that total and complexed PSA concentrations failed to discriminate between benign and malignant disease. In contrast, the areas under the ROC curve were greater for the free-to-total ratio (P = 0.033). CONCLUSION: These results show that in patients with total PSA concentrations between 2 and 10 micro g/L, the free-to-total PSA ratio was superior to total PSA concentration in discriminating between patients with benign and malignant disease.

HER2/neu gene amplification and protein overexpression in G3 pT2 transitional cell carcinoma of the bladder: a role for anti-HER2 therapy?

HER2/neu is an oncogene encoding a type 1 tyrosine kinase growth factor receptor. Polysomy 17, gene amplification and HER2/neu protein overexpression are associated with a poor prognosis in transitional cell carcinomas (TCC) of the bladder. Due to the application of different laboratory techniques, the exact incidence of HER/neu abnormalities remains uncertain in TCC. Standardised laboratory techniques are therefore important in the determination of the HER2/neu status if an assessment of the potential value of anti-HER2/neu treatments in the clinical management of patients with TCC is to be made. In this study, 75 TCCs with evidence of detrusor muscle invasion at first clinical presentation were included. Gene amplification, polysomy 17 and HER2 copy number were assessed using fluorescence in situ hybridisation (FISH), with separate probes for chromosome 17 and HER2/neu. Protein overexpression was assessed using immunohistochemistry (IHC), with the CB11 antibody and a scoring system evaluating only membranous staining as positive. The mean patient age was 69.5 years (range 42-93 years) and the median survival was 15 months (range 1-156 months). Polysomy 17 occurred in 97%, increased HER/neu copy number in 92% and HER2/neu gene amplification in 7%. Protein overexpression occurred in 57% of cases. Polysomy 17 and HER2/neu protein overexpression are common in G3 pT2 TCCs of the bladder. However, gene amplification is uncommon. Mechanisms other than gene amplification may be responsible for protein overexpression in this tumour type. Evidence from breast cancer suggests that only tumours with HER2/neu gene amplification respond to the anti-HER2/neu therapy trastuzumab (Herceptin). If this were true for bladder cancer, only 4/75 (5%) of G3 pT2 TCCs would be suitable for treatment. The role of trastuzumab in these tumours remains untested at present.

HER2/neu overexpression in the development of muscle-invasive transitional cell carcinoma of the bladder.

The mortality from transitional cell carcinoma (TCC) of the urinary bladder increases significantly with the progression of superficial or locally invasive disease (pTa/pT1) to detrusor muscle-invasive disease (pT2+). The most common prognostic markers in clinical use are tumour stage and grade, which are subject to considerable intra- and interobserver variation. Polysomy 17 and HER2/neu gene amplification and protein overexpression have been associated with more advanced disease. Standardised techniques of fluorescence in situ hybridisation and immunohistochemistry, which are currently applied to other cancers with a view to offering anti-HER2/neu therapies, were applied to tumour pairs comprising pre- and postinvasive disease from 25 patients undergoing treatment for bladder cancer. In the preinvasive tumours, increased HER2/neu copy number was observed in 76% of cases and increased chromosome 17 copy number in 88% of cases, and in the postinvasive group these values were 92 and 96%, respectively (not significantly different P=0.09 and 0.07, respectively). HER2 gene amplification rates were 8% in both groups. Protein overexpression rates were 76 and 52%, respectively, in the pre- and postinvasive groups (P=0.06). These results suggest that HER2/neu abnormalities occur prior to and persist with the onset of muscle-invasive disease. Gene amplification is uncommon and other molecular mechanisms must account for the high rates of protein overexpression. Anti-HER2/neu therapy might be of use in the treatment of TCC.