Stimulated Raman Histology Interpretation by Artificial Intelligence Provides Near-Real-Time Pathologic Feedback for Unprocessed Prostate Biopsies.

PURPOSE: Stimulated Raman histology is an innovative technology that generates real-time, high-resolution microscopic images of unprocessed tissue, significantly reducing prostate biopsy interpretation time. This study aims to evaluate the ability for an artificial intelligence convolutional neural network to interpretate prostate biopsy histologic images created with stimulated Raman histology. MATERIALS AND METHODS: Unprocessed, unlabeled prostate biopsies were prospectively imaged using a stimulated Raman histology microscope. Following stimulated Raman histology creation, the cores underwent standard pathological processing and interpretation by at least 2 genitourinary pathologists to establish a ground truth assessment. A network, trained on 303 prostate biopsies from 100 participants, was used to measure the accuracy, sensitivity, and specificity of detecting prostate cancer on stimulated Raman histology relative to conventional pathology. The performance of the artificial intelligence was evaluated on an independent 113-biopsy test set. RESULTS: Prostate biopsy images obtained through stimulated Raman histology can be generated within a time frame of 2 to 2.75 minutes. The artificial intelligence system achieved a rapid classification of prostate biopsies with cancer, with a potential identification time of approximately 1 minute. The artificial intelligence demonstrated an impressive accuracy of 96.5% in detecting prostate cancer. Moreover, the artificial intelligence exhibited a sensitivity of 96.3% and a specificity of 96.6%. CONCLUSIONS: Stimulated Raman histology generates microscopic images capable of accurately identifying prostate cancer in real time, without the need for sectioning or tissue processing. These images can be interpreted by artificial intelligence, providing physicians with near-real-time pathological feedback during the diagnosis or treatment of prostate cancer.

Vulnerable newborn types: analysis of subnational, population-based birth cohorts for 541 285 live births in 23 countries, 2000-2021.

OBJECTIVE: To examine prevalence of novel newborn types among 541 285 live births in 23 countries from 2000 to 2021. DESIGN: Descriptive multi-country secondary data analysis. SETTING: Subnational, population-based birth cohort studies (n = 45) in 23 low- and middle-income countries (LMICs) spanning 2000-2021. POPULATION: Liveborn infants. METHODS: Subnational, population-based studies with high-quality birth outcome data from LMICs were invited to join the Vulnerable Newborn Measurement Collaboration. We defined distinct newborn types using gestational age (preterm [PT], term [T]), birthweight for gestational age using INTERGROWTH-21st standards (small for gestational age [SGA], appropriate for gestational age [AGA] or large for gestational age [LGA]), and birthweight (low birthweight, LBW [<2500 g], nonLBW) as ten types (using all three outcomes), six types (by excluding the birthweight categorisation), and four types (by collapsing the AGA and LGA categories). We defined small types as those with at least one classification of LBW, PT or SGA. We presented study characteristics, participant characteristics, data missingness, and prevalence of newborn types by region and study. RESULTS: Among 541 285 live births, 476 939 (88.1%) had non-missing and plausible values for gestational age, birthweight and sex required to construct the newborn types. The median prevalences of ten types across studies were T+AGA+nonLBW (58.0%), T+LGA+nonLBW (3.3%), T+AGA+LBW (0.5%), T+SGA+nonLBW (14.2%), T+SGA+LBW (7.1%), PT+LGA+nonLBW (1.6%), PT+LGA+LBW (0.2%), PT+AGA+nonLBW (3.7%), PT+AGA+LBW (3.6%) and PT+SGA+LBW (1.0%). The median prevalence of small types (six types, 37.6%) varied across studies and within regions and was higher in Southern Asia (52.4%) than in Sub-Saharan Africa (34.9%). CONCLUSIONS: Further investigation is needed to describe the mortality risks associated with newborn types and understand the implications of this framework for local targeting of interventions to prevent adverse pregnancy outcomes in LMICs.

Differential 18F-NaF uptake in various compartments in knee osteoarthritis: an observational study using PET/MRI.

AIM: To investigate if the pattern of fluorine-18-labelled sodium fluoride (18F-NaF) uptake on integrated positron-emission tomography (PET)/magnetic resonance imaging (MRI) of bone marrow lesions (BML) and osteophytes differs between different knee compartments. MATERIALS AND METHODS: Sixteen patients with no prior history of knee injury with or without pain were recruited for the study. The images of both knees were acquired on simultaneous PET/MRI. The acquisition was done after 45 minutes of intravenous injection of 18F-NaF 185-370 MBq (5-10 mCi) for 40 minutes. Each knee was divided into eight compartments patella, trochlea, medial central femur, lateral central femur, medial posterior femur, lateral posterior femur, medial tibia, lateral tibia, and cruciate ligament insertion specifically for BML. BML and osteophytes were scored using MRI Osteoarthritis Knee Score (MOAKS) criteria and their corresponding maximum standardised uptake values (SUVmax) recorded. RESULTS: BML and osteophytes both showed statistically significant differences among knee compartments, i.e., p-value <0.000 and < 0.043 respectively. SUVmax for BML and osteophytes was greatest in the medial tibia. CONCLUSION: 18F-NaF PET/MRI showed that BML and osteophytes had differential uptake values due to bone remodelling amongst the various knee compartments and this may help to design disease-modifying osteoarthritis drugs in the future.

Oral health status and absence from school among 12 year olds.

OBJECTIVE: To assess dental caries, periodontal status, malocclusion and absenteeism from school among 12-year-olds in Bhopal district, Central India. MATERIALS AND METHODS: Two-stage random sample of 1238 school children. Decayed missing filled teeth (DMFT), Significant caries index (SiC), community periodontal index (CPI) and dental aesthetic index (DAI) were used to record dental caries, periodontal status and malocclusion. Information on absence from school in the previous year due to pain/discomfort in the teeth or mouth was collected via interviews. Generalized structural equation modelling (GSEM) examined the direct and indirect predictors of absence from school. RESULTS: A total of 39.1%, 17.3% and 23.9% of children had dental caries, calculus and gingival bleeding respectively. Mean DMFT and SiC scores were 1.82± 1.36 and 3.15 ± 1.47. 5,127 school hours were missed due to oral health problems per 1,000 children. None of the studied variables predicted absence from school. Utilization of dental care was associated directly with gender and malocclusion (p⟨ 0.001). Periodontal status was associated with male gender, nuclear families, tobacco consumption, and malocclusion (p⟨ 0.001). Higher DMFT was associated with male gender, malocclusion and experience of pain/discomfort (p⟨ 0.001). CONCLUSIONS: Poor oral health and a high prevalence of untreated dental caries were noted. Despite a considerable number of missed school hours reported due to dental conditions, none of the studied variables predicted absence from school.

TOOKAD® Soluble focal therapy: pooled analysis of three phase II studies assessing the minimally invasive ablation of localized prostate cancer.

PURPOSE: To evaluate the 6-month effects of the recommended drug and light dosage in focal vascular-targeted photodynamic therapy (VTP) using TOOKAD(®) Soluble in patients with localized prostate cancer (LPCa). METHODS: We performed a pooled analysis of 117 men with LPCa, PSA <10 ng/mL, and Gleason score ≤ 7 (3 + 4), from 3 studies who received a 10-min intravenous infusion of a single dose of 4 mg/kg TOOKAD(®) Soluble, activated by a 753-nm light at 200 J/cm delivered in the prostate by transperineal fibres under transrectal ultrasound guidance. Primary endpoint was 6-month negative biopsies in the treated lobe(s). PSA was measured at month 1, 3, and 6. Magnetic resonance imaging was performed at day 7, month 3, and 6. International Prostate Symptom Score (IPSS), International Index of Erectile Function (IIEF-5) and adverse events were reported at day 7, month 1, 3, and 6. RESULTS: Month 6 negative biopsy rate was 68.4 % in the overall evaluable population (N = 114) and 80.6 % for patients treated by hemiablation with light density index (LDI) ≥ 1 (N = 67). Mean prostate necroses at week-1 were 76.5 and 86.3 %, respectively. In both groups, PSA levels at month 6 decreased by 2.0 ng/mL. Small changes from baseline for IPSS and IIEF-5 indicated a slight improvement in urinary function and a slight deterioration in sexual function. CONCLUSIONS: Focal VTP treatment with TOOKAD(®) Soluble at 4 mg/kg and 200 J/cm resulted in a negative 6-month biopsy rate of 68.4 % for the whole population and 80.6 % for patients treated by hemiablation with LDI ≥ 1. The treatment was well tolerated. Two phase III studies will reach completion in early 2015.

Does normalisation improve the diagnostic performance of apparent diffusion coefficient values for prostate cancer assessment? A blinded independent-observer evaluation.

AIM: To evaluate the performance of normalised apparent diffusion coefficient (ADC) values for prostate cancer assessment when performed by independent observers blinded to histopathology findings. MATERIALS AND METHODS: Fifty-eight patients undergoing 3 T phased-array coil magnetic resonance imaging (MRI) including diffusion-weighted imaging (DWI; maximal b-value 1000 s/mm(2)) before prostatectomy were included. Two radiologists independently evaluated the images, unaware of the histopathology findings. Regions of interest (ROIs) were drawn within areas showing visually low ADC within the peripheral zone (PZ) and transition zone (TZ) bilaterally. ROIs were also placed within regions in both lobes not suspicious for tumour, allowing computation of normalised ADC (nADC) ratios between suspicious and non-suspicious regions. The diagnostic performance of ADC and nADC were compared. RESULTS: For PZ tumour detection, ADC achieved significantly higher area under the receiver operating characteristic curve (AUC; p=0.026) and specificity (p=0.021) than nADC for reader 1, and significantly higher AUC (p=0.025) than nADC for reader 2. For TZ tumour detection, nADC achieved significantly higher specificity (p=0.003) and accuracy (p=0.004) than ADC for reader 2. For PZ Gleason score >3+3 tumour detection, ADC achieved significantly higher AUC (p=0.003) and specificity (p=0.005) than nADC for reader 1, and significantly higher AUC (p=0.023) than nADC for reader 2. For TZ Gleason score >3+3 tumour detection, ADC achieved significantly higher specificity (p=0.019) than nADC for reader 1. CONCLUSION: In contrast to prior studies performing unblinded evaluations, ADC was observed to outperform nADC overall for two independent observers blinded to the histopathology findings. Therefore, although strategies to improve the utility of ADC measurements in prostate cancer assessment merit continued investigation, caution is warranted when applying normalisation to improve diagnostic performance in clinical practice.

Assessment of change in prostate volume and shape following surgical resection through co-registration of in-vivo MRI and fresh specimen ex-vivo MRI.

AIM: To compare the size and shape of the prostate between in-vivo and fresh ex-vivo magnetic resonance imaging (MRI), in order to quantify alterations in the prostate resulting from surgical resection. MATERIAL AND METHOD: Ten patients who had undergone 3 T prostate MRI using a phased-array coil and who were scheduled for prostatectomy were included in this prospective study. The ex-vivo specimen underwent MRI prior to formalin fixation or any other histopathological processing. Prostate volume in vivo and ex vivo was assessed using planimetry. Prostate shape was assessed by calculating ratios between the diameters of the prostate in all three dimensions. RESULTS: Mean prostate volume was significantly smaller ex vivo than in vivo (39.7 ± 18.6 versus 50.8 ± 26.8 cm(3); p = 0.008), with an average change in volume of -19.5%. The right-to-left (RL)/anteroposterior (AP) ratio of the prostate, representing the shape of the prostate within its axial plane, was significantly larger ex vivo than in vivo (1.33 ± 0.14 versus 1.21 ± 0.12; p = 0.015), with an average percent change in RL/AP ratio of the prostate of +12.2%. There was no significant difference between in-vivo and ex-vivo acquisitions in terms of craniocaudal (CC)/AP (p = 0.963, median change = -2.1%) or RL/CC (p = 0.265, median change = +1.3%) ratios. CONCLUSION: The observed volume and shape change following resection has not previously been assessed by comparison of in-vivo and fresh ex-vivo MRI and likely represents loss of vascularity and of connective tissue attachments in the ex-vivo state. These findings have implications for co-registration platforms under development to facilitate improved understanding of the accuracy of MRI in spatial localization of prostate tumours.

Synthesis of single-enantiomer bioactive molecules: a brief overview.

Chiral-center enantiomers have been shown to differ significantly in biological activity, pharmacodynamics, pharmacokinetics and toxicity. New developments in the stereoselective organic synthesis have enriched the vast literature of synthetic methodologies applicable to access natural products as well as bioactive molecules. These compounds also include new drugs, drug candidates and reagents used to explore biological processes. The article reviews the synthesis of optically pure drugs, biologically active intermediates and amino alcohols by using different methods.

Descriptive epidemiology of rotavirus infection in a community in North India.

In India, rotavirus infections cause the death of 98621 children each year. In urban neighbourhoods in Delhi, children were followed up for 1 year to estimate the incidence of rotavirus gastroenteritis and common genotypes. Infants aged f1 week were enrolled in cohort 1 and infants aged 12 months (up to +14 days) in cohort 2. Fourteen percent (30/210) gastroenteritis episodes were positive for rotavirus. Incidence rates of rotavirus gastroenteritis episodes in the first and second year were 0.18 [95% confidence interval (CI) 0.10­0.27] and 0.14 (95% CI 0.07­0.21) episodes/child-year, respectively. The incidence rate of severe rotavirus gastroenteritis in the first year of life was 0.05 (95% CI 0.01­0.10) episodes/child-year. There were no cases in the second year. The common genotypes detected were G1P[8] (27%) and G9P[4] (23%). That severe rotavirus gastroenteritis is common in the first year of life is relevant for planning efficacy trials.

Preliminary experience with a novel method of three-dimensional co-registration of prostate cancer digital histology and in vivo multiparametric MRI.

AIM: To assess a novel method of three-dimensional (3D) co-registration of prostate cancer digital histology and in-vivo multiparametric magnetic resonance imaging (mpMRI) image sets for clinical usefulness. MATERIAL AND METHODS: A software platform was developed to achieve 3D co-registration. This software was prospectively applied to three patients who underwent radical prostatectomy. Data comprised in-vivo mpMRI [T2-weighted, dynamic contrast-enhanced weighted images (DCE); apparent diffusion coefficient (ADC)], ex-vivo T2-weighted imaging, 3D-rebuilt pathological specimen, and digital histology. Internal landmarks from zonal anatomy served as reference points for assessing co-registration accuracy and precision. RESULTS: Applying a method of deformable transformation based on 22 internal landmarks, a 1.6 mm accuracy was reached to align T2-weighted images and the 3D-rebuilt pathological specimen, an improvement over rigid transformation of 32% (p = 0.003). The 22 zonal anatomy landmarks were more accurately mapped using deformable transformation than rigid transformation (p = 0.0008). An automatic method based on mutual information, enabled automation of the process and to include perfusion and diffusion MRI images. Evaluation of co-registration accuracy using the volume overlap index (Dice index) met clinically relevant requirements, ranging from 0.81-0.96 for sequences tested. Ex-vivo images of the specimen did not significantly improve co-registration accuracy. CONCLUSION: This preliminary analysis suggests that deformable transformation based on zonal anatomy landmarks is accurate in the co-registration of mpMRI and histology. Including diffusion and perfusion sequences in the same 3D space as histology is essential further clinical information. The ability to localize cancer in 3D space may improve targeting for image-guided biopsy, focal therapy, and disease quantification in surveillance protocols.

Effect of laser bleaching, ultrasonic scaling and powered tooth brushing on surface roughness and bacterial adherence of class V composite restorations.

Objective: To evaluate and compare the effect of diode laser assisted bleaching, ultrasonic scaling and powered tooth brushing on surface roughness and bacterial adherence on class V cavities restored with composites. Materials and methods: A total of one hundred and twenty samples (40 samples each of Brilliant Everglow, Beautifil II and Heytec-N) were prepared in standardized stainless steel molds. The samples were further subdivided into four subgroups i.e. one control group (without any intervention) and three experimental groups - diode laser assisted bleaching, ultrasonic scaling and powered tooth brushing consisting of 10 sample each. Surface roughness was measured quantitatively with the help of 3D Optical Profilometer. For bacterial adherence analysis S. mutans strain (ATCC 25175) was cultured in BHI medium and samples were evaluated for the presence of viable bacteria using the Colony Forming Unit (CFU) count. Results obtained were then tabulated and subjected to statistical analysis. Results: Diode laser bleaching caused a significant increase in surface roughness and bacterial adherence with lowest mean change exhibited by Heytec-N followed by Beautifil II and highest by Brilliant Everglow group. Similarly, Ultrasonic scaling increased the surface roughness of all the three tested samples with significant difference between the groups. Powered tooth brushing had no effect on the surface roughness and bacterial adherence of the tested composites. Conclusion: Diode assisted laser bleaching and ultrasonic caused significantly higher surface roughness and bacterial adherence values for all the tested composites. It may therefore be recommended to do finishing and polishing of restorations after such procedures.

Impact of delay after biopsy and post-biopsy haemorrhage on prostate cancer tumour detection using multi-parametric MRI: a multi-reader study.

AIM: To assess impact of haemorrhage and delay after biopsy on prostate tumour detection using multi-parametric (MP) magnetic resonance imaging (MRI) assessment. MATERIALS AND METHODS: Forty-four patients underwent prostate MRI at 1.5 T using a pelvic phased-array coil, including T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), and dynamic contrast-enhanced (DCE) imaging, before prostatectomy. Three radiologists independently reviewed images during four sessions [T2WI, DWI, DCE, and all parameters combined (MP-MRI)] to assess for tumour in each sextant. In a separate session, readers reviewed T1WI to score the extent of haemorrhage per sextant. Accuracy was assessed using logistic regression for correlated data. RESULTS: There was no significant difference in accuracy between readers for any session (p ≥ 0.166), and results were averaged across the three readers for remaining comparisons. Accuracy was significantly greater for MP-MRI than for any parameter alone (p ≤ 0.020). For T2WI alone, there was a trend toward decreased sensitivity in sextants with extensive haemorrhage (p = 0.072). However, accuracy, sensitivity, and specificity were otherwise similar for sextants with and without extensive haemorrhage for all sessions (p = 0.192-0.934). No session showed a significant improvement in accuracy, sensitivity, or specificity in cases with delay after biopsy of over 4 weeks compared with shorter delay. CONCLUSION: Extensive haemorrhage and short delay after biopsy did not negatively impact accuracy for tumour detection using MP-MRI. Further studies using MP-MRI protocols and interpretation schemes from other institutions are required to confirm these observations.

Metabonomic analysis of hepatitis E patients shows deregulated metabolic cycles and abnormalities in amino acid metabolism.

Hepatitis E, which is endemic to resource-poor regions of the world, is largely an acute and self-limiting disease, but some patients have an increased susceptibility to develop fulminant hepatitis. The pathogenesis of hepatitis E in humans is poorly characterized. To understand the metabolic pathways involved in the pathophysiology of hepatitis E, we have used (1) H nuclear magnetic resonance spectroscopy to quantify various metabolites in the plasma and urine of the patients with hepatitis E. These were compared with specimens from patients with acute hepatitis B as disease controls and healthy volunteers. Data were analysed using chemometric statistical methods and metabolite databases. The main metabonomic changes found in patients with hepatitis E, but not in those with hepatitis B, included increased plasma levels of L-isoleucine, acetone, and glycerol, reduced plasma levels of glycine, and reduced urinary levels of imidazole, 3-aminoisobutanoic acid, 1-methylnicotinamide, biopterin, adenosine, 1-methylhistidine, and salicyluric acid. Patients with hepatitis E or B both showed increased levels of plasma and urinary L-proline and decreased levels of various other metabolites. Pathway analysis tools suggest the involvement of glycolysis, tricarboxylic acid cycle, urea cycle, and amino acid metabolism in patients with acute hepatitis E. These findings may help better understand the clinical and biochemical manifestations in this disease and the underlying pathophysiologic processes. Based on our findings, it would be worthwhile determining whether patients with hepatitis E are more prone to develop lactic acidosis and ketosis compared with other forms of viral hepatitis.

Prediction of sustained virological response to combination therapy with pegylated interferon alfa and ribavirin in patients with genotype 3 chronic hepatitis C.

BACKGROUND: Sustained virological response (SVR) rates in patients with hepatitis C are heterogeneous and are influenced by a wide range of host and viral factors. AIM: To evaluate the efficacy of combination therapy with pegylated interferon alfa (PEG-IFN-α) and ribavirin (RBV), and document the SVR rates taking into consideration various predictive factors in patients with chronic hepatitis C (CHC) genotype 3. METHODS: Ninety-seven treatment-naive patients with CHC genotype 3 (mean age 41.46±11.51 years, M:F ratio 79:18), who received a combination of PEG-IFN (α-2a or α-2b) and RBV were retrospectively analyzed (2006-2008) for the early virological response (EVR) at 12 weeks, end of treatment response (ETR), and SVR at 6 months. RESULTS: Eighty-four (86.6%) patients achieved EVR and 81 (83.5%) achieved ETR, while SVR was achieved in 65 (67.0%) patients. Of the 84 patients who achieved EVR, 77 (91.7%) achieved ETR and 61 (72.6%) achieved SVR at 6 months. Age and body mass index (BMI) were found to be important predictors (*P<0.05) of SVR. CHC patients with a history of alcohol intake showed decreased SVR (52%) (*P=0.035) as compared to nonalcoholics (80%). Cirrhotic versus noncirrhotic patients showed no difference in SVR (54.5% vs. 70.7%) (P=0.157). Serum alanine aminotransferase (ALT) (P=0.169) and hepatitis C virus (HCV) RNA levels (P=0.42) also did not have an influence on the SVR. CONCLUSION: Combination therapy with PEG-IFN-α and RBV demonstrated good tolerability in CHC genotype 3 infection. Age, BMI, and alcohol consumption play an important role in determining treatment outcome.

What is a better modality of maxillomandibular fixation: bone-supported arch bars or Erich arch bars? A systematic review and meta-analysis.

Any procedure that aims to manage maxillofacial fracture is incomplete without meticulous maxillomandibular fixation (MMF). For decades, Erich arch bars (EABs) have been used for this purpose, but with the advent of bone-supported arch bars (BSABs), more surgeons now prefer them to conventional EABs. The present study was designed to identify which of the two methods is best. An exhaustive literature search was conducted in June 2020 on various electronic databases to select studies that compared EABs and BSABs. Outcomes such as duration of placement, stability, oral hygiene, and complications such as damage to the roots of teeth and needle-stick injury, were analysed. A total of 716 studies were identified, of which seven were eligible for inclusion. The meta-analysis showed that the use of BSABs is significantly faster with no needle penetration and better oral hygiene. Both arch bars are equally stable, but root damage is an associated complication. The available literature to date shows that BSABs are a better option than EABs. However, further research is recommended, as these studies are associated with various confounding factors.

Endoscopic ultrasound-guided fine-needle aspiration cytology in the evaluation of suspected tuberculosis in patients with isolated mediastinal lymphadenopathy.

BACKGROUND AND STUDY AIMS: Patients with suspected tuberculosis without pulmonary lesions and with mediastinal lymphadenopathy often pose a diagnostic challenge. Endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) cytology is an established modality to evaluate mediastinal and abdominal lesions. The aim of the present study was to evaluate the role of EUS-FNA in isolated mediastinal lymphadenopathy in patients suspected of having tuberculosis. METHODS: Consecutive patients suspected of having tuberculosis with isolated mediastinal lymphadenopathy were included in a prospective study. Mediastinal lymphadenopathy was diagnosed on a contrast-enhanced computed tomography scan of the chest. Patients with concomitant lung parenchymal lesions were excluded. Previous attempts to diagnose the etiology of lymphadenopathy had failed in 69 % of patients. EUS-FNA was performed on an outpatient basis under conscious sedation. The sensitivity, specificity, and diagnostic accuracy of EUS-FNA were calculated. RESULTS: A total of 60 consecutive patients (mean age 39.8 years, 58 % males) with mediastinal lymphadenopathy were included. EUS confirmed the presence of mediastinal lymph nodes ranging in size from 8 mm to 40 mm (mean 26 mm) in all patients. EUS-FNA provided an adequate tissue sample in 54 patients during the first examination and repeat EUS-FNA was necessary in six patients. A final diagnosis was obtained by EUS-FNA in 42 patients (tuberculosis in 32, sarcoidosis in six, and Hodgkin's disease in four patients). An additional 14 patients were treated for tuberculosis based on EUS-FNA and clinical features. Mediastinoscopy was required for diagnosis in the remaining four patients. EUS-FNA had an overall diagnostic yield of 93 %, sensitivity of 71 %, specificity of 100 %, and positive predictive value of 100 %. CONCLUSION: EUS-FNA is an accurate, safe, and minimally invasive modality for evaluating isolated mediastinal lymphadenopathy in patients suspected of having tuberculosis in an endemic area with a high prevalence of tuberculosis.

Site-specific regulation of CA(V)2.2 channels by protein kinase C isozymes betaII and epsilon.

Ca(v)2.2 high voltage-gated calcium channels are regulated by phorbol-12-myristae, 13-acetate (PMA) via Ser/Thr protein kinase C (PKC) phosphorylation sites in the I-II linker and C-terminus of the alpha(1) 2.2 subunit. Here we show that PMA enhancement of Ca(v)2.2 currents expressed in Xenopus oocytes can be blocked by inhibitors of PKC betaII or PKC epsilon isozymes, as shown previously for Ca(v)2.3 currents, and that microinjection of PKC betaII or PKC epsilon isozymes in the oocytes expressing the WT Ca(v)2.2 channels increases the basal barium current (I(Ba)). The I-V plot shows a large increase in current amplitude with PKC betaII and PKC epsilon isozymes with only a small shift in the peak I(Ba) in the hyperpolarizing direction. The potentiation of Ca(v)2.2 currents by microinjection of PKC betaII and PKC epsilon isozymes was not altered by the inhibition of G proteins with GDPbetaS. The combination of isozyme specific inhibitors with previously generated Ser/Thr to Ala mutants of alpha(1) 2.2 subunit revealed that PKC betaII or PKC epsilon isozymes (but not PKC alpha or delta) can provide full enhancement through the stimulatory site (Thr-422) in the I-II linker but that PKC epsilon is better at decreasing channel activity through the inhibitory site Ser-425. The enhancing effect of PKC betaII or epsilon at Thr-422 is dominant over the inhibitory effect at Ser-425. Injected PKC betaII also enhances Ca(v)2.2 current when any of the potential stimulatory sites (Ser-1757, Ser-2108 and Ser-2132) are available in the C-terminus. PKC epsilon provides lesser enhancement with C-terminal sites and only with Ser-2108 and Ser-2132. Sites Ser-1757 and Ser-2132, but not Ser-2108, are dominant over the inhibitory site Ser-425. Collectively, these results reveal a hierarchy of regulatory sites in Ca(v)2.2 channels. Site-specific regulation by different PKC isozymes may allow graded levels of channel activation and susceptibility or resistance to subsequent stimulatory events.

Sonographic correlations with the new molecular classification of invasive breast cancer.

Ultrasound is a useful adjunct to mammography for the characterisation and biopsy of solid breast lesions. Protein expression profiling of breast cancer has identified specific subgroups with potential clinical, biological and therapeutic implications. The aim of this study was to determine the ultrasound correlates of these novel molecular classes of invasive breast cancer. The ultrasound findings in 358 patients with operable breast cancer were correlated with the previously described protein expression classes identified by our group using immunohistochemical (IHC) assessment of a large series of breast cancer cases in which 25 proteins of known relevance in breast cancer were assessed, including hormone receptors, HER2 status, basal and luminal markers, p53 and e-cadherin. The proportion of occult lesions was not significantly different in the two groups. Significant differences were noted between the two groups expressing luminal epithelial markers and hormone receptors (1 and 2), including a greater proportion of ill-defined, irregular and distally attenuating tumours in group 2. Tumours characterised by c-erbB2/MUC1 expression, with weak hormone receptor positivity (group 3) were also more likely to be ill defined. Tumours expressing basal markers (group 5) were less likely to have an echogenic halo. The ultrasound features of breast cancer show areas of significant correlation with molecular classes of invasive breast cancer identified by IHC analysis. The biological reasons for these findings and their implications regarding imaging protocols require further study and may enable improved detection of these lesions.