Carboplatin enhances lymphocyte-endothelial interactions to promote CD8+ T cell trafficking into the ovarian tumor microenvironment.

OBJECTIVES: Standard chemotherapy agents, including carboplatin, have known immunogenic properties. We sought to determine how carboplatin may influence lymphocyte trafficking to tumor sites. METHODS: Murine models of ovarian cancer were utilized to examine lymphocyte trafficking with common clinically used agents including carboplatin, anti-PD-1 antibody, or anti-VEGFR-2 antibody. Adhesion interactions of lymphocytes with tumor vasculature were measured using intravital microscopy, lymphocyte homing with immunohistochemistry, and treatment groups followed for overall survival. RESULTS: Carboplatin chemotherapy profoundly alters the tumor microenvironment to promote lymphocyte adhesive interactions with tumor vasculature and resultant improvement in lymphocyte trafficking. The measured results seen with carboplatin in the tumor microenvironment were superior to anti-PD-1 treatment or anti-VEGFR-2 which may have contributed to increased overall survival in carboplatin treated groups. CONCLUSIONS: These novel findings suggest a role for chemotherapeutic agents to broadly influence anti-tumor immune responses beyond the induction of immunogenic tumor cell death.

Severity of intervertebral disc herniation regulates cytokine and chemokine levels in patients with chronic radicular back pain.

OBJECTIVE: The contributions of intervertebral disc disease and subject-specific covariates to systemic inflammation in low back pain are unknown. We examined the effects of symptomatic disc herniation (DH) and MRI herniation severity on serum cytokine levels in clinical subjects. DESIGN: Cytokine levels from lumbar DH subjects (N = 78) were compared to control subjects (N = 57) accounting for effects of DH, age, body mass index (BMI) and gender. Effect of DH severity on cytokine levels was analyzed on subsets of subjects with acute or chronic pain. Serum cytokines were also analyzed in a subset of patients between pre- and 3 months post-surgery. RESULTS: Cytokine levels were elevated in the serum of patients with symptomatic DH, and the covariates age, BMI and gender significantly contributed to levels of some cytokines. Severity of herniation was a significant contributor to pain intensity (VAS), serum levels of HMGB1, PDGFbb, and IL-9. The relationship between DH severity and cytokine levels was confirmed in subjects with chronic, but not acute symptoms. Serum levels of macrophage migration inhibitory factor (MIF) decreased, whereas levels of CCL3, CCL11, CXCL1, and CXCL10 were significantly elevated post surgery. CONCLUSIONS: This study is the first to show that DH severity is coordinately associated with changes in serum levels of inflammatory cytokines in chronic pain subjects. HMGB1, PDGFbb and IL-9 are novel mediators of increasing DH severity, indicative of cellular damage, neuro-inflammation and angiogenesis. Resolution of inflammation was observed with decrease in MIF post surgery. However, elevated chemokine levels indicate ongoing remodeling and wound healing at 3-month time point.

Mitochondrial DNA in the tumour microenvironment activates neutrophils and is associated with worse outcomes in patients with advanced epithelial ovarian cancer.

BACKGROUND: Advanced cancer causes necrosis and releases damage-associated molecular patterns (DAMPs). Mitochondrial DAMPs activate neutrophils, including generation of neutrophil extracellular traps (NETs), which are injurious, thrombogenic, and implicated in metastasis. We hypothesised that extracellular mitochondrial DNA (mtDNA) in ascites from patients with epithelial ovarian cancer (EOC) would correlate with worse outcomes. METHODS: Banked ascites supernatants from patients with newly diagnosed advanced EOC were analysed for mtDNA, neutrophil elastase, and activation of healthy donor neutrophils and platelets. TCGA was mined for expression of SELP and ELANE. RESULTS: The highest quartile of ascites mtDNA correlated with reduced progression-free survival (PFS) and a higher likelihood of disease progression within 12-months following primary surgery (n = 68, log-rank, p = 0.0178). NETs were detected in resected tumours. Ascites supernatants chemoattracted neutrophils, induced NETs, and activated platelets. Ascites exposure rendered neutrophils suppressive, based on abrogation of ex vivo stimulated T cell proliferation. Increased SELP mRNA expression correlated with worse overall survival (n = 302, Cox model, p = 0.02). CONCLUSION: In this single-centre retrospective analysis, ascites mtDNA correlated with worse PFS in advanced EOC. Mitochondrial and other DAMPs in ascites may activate neutrophil and platelet responses that facilitate metastasis and obstruct anti-tumour immunity. These pathways are potential prognostic markers and therapeutic targets.

Impact of ascites volume on clinical outcomes in ovarian cancer: A cohort study.

OBJECTIVES: To investigate the impact of ascites volume on ovarian cancer outcomes. METHODS: Clinicopathologic features of a cohort of patients with ovarian cancer were obtained from a curated database at a single institution. Progression free survival (PFS) and overall survival (OS) were recorded. Ascites volume at primary surgery was dichotomized at 2000mL and comparisons for high and low volume ascites were made. Additionally, to elucidate interactions between ascites and ovarian tumor progression, we evaluated the effect of intraperitoneal administrations of murine cell-free ascites versus saline in a syngeneic mouse model of epithelial ovarian cancer. RESULTS: Out of 685 patients identified, 58% had ascites present at the time of initial surgery. Considering the volume of ascites continuously, each liter of ascites was associated with shorter PFS (HR=1.12, 95% CI: 1.07-1.17) and OS (HR=1.12, 95%CI: 1.07-1.17). Patients with ascites greater than the median of 2000mL had significantly shorter PFS (14.5months vs. 22.7months; p<0.001) and OS (27.7months vs. 42.9months; p<0.001). After adjusting for stage, presence of ascites was inversely associated with ability to achieve optimal cytoreductive surgery. Consistent with these correlative results in patients, intraperitoneal administrations of murine cell-free ascites accelerated ovarian cancer progression in mice. CONCLUSIONS: The volume of ascites at initial diagnosis of ovarian cancer correlated with worse PFS and OS. The effect of large volume on prognosis is likely to be in part related to reduced likelihood for complete resection of tumor (R0). If these findings are confirmed in independent studies, consideration should be made to add the presence of large volume ascites at diagnosis to the staging criteria for ovarian cancer.

Transcultural adaptation of the Breast Cancer Awareness Measure.

AIM: To overcome the lack of a validated and robust Arabic instrument to measure breast cancer awareness. BACKGROUND: Currently, there is no validated Arabic instrument for measuring breast cancer awareness levels. We adapted, translated and validated the Breast Cancer Awareness Measure developed by Cancer Research UK. METHODS: The instrument was translated into Arabic and back-translated for validation. Validation and reliability tests were conducted using purposively sampled 972 Arab women older than 20 years, living in Oman. The adapted content was validated by a panel of medical, linguistic and cultural experts, followed by cognitive interviews (n = 10), behavioural coding (n = 30) and criterion validation (n = 646). The instrument was tested for acceptability and its subscales for internal consistency. Inter-rater reliability was estimated between two similar groups (n = 144 and n = 142) to test homogeneity. RESULTS: The adapted and translated instrument had a high acceptability (98.7% completed). The validation process shaped the adaptation, and resulted in strong criterion validity (R = 0.58, P < 0.01). The instrument subscales for risk factors and warning signs had high internal consistency (Cronbach's alpha 0.856 and 0.890, respectively), with all floor and ceiling effects less than 15%. The correlation measure for inter-rater reliability was 0.97 (P < 0.01). DISCUSSION: Through the incorporation of contextual characteristics and prevalent beliefs among Arab populations, the adapted Best Cancer Awareness Measure is a robust Arabic instrument for the measurement of breast cancer awareness and early detection practices among Arab women. LIMITATIONS: The purposively selected sample may not be representative of the population. CONCLUSION AND IMPLICATIONS FOR HEALTH POLICY: Improvement of awareness and early detection of breast cancer can contribute towards reducing mortality from the disease. The adapted instrument has policy implications, since measurement of awareness levels is essential towards breast health promotion policies in Arab countries.

The investigational agent MLN2238 induces apoptosis and is cytotoxic to CLL cells in vitro, as a single agent and in combination with other drugs.

Chronic lymphocytic leukaemia (CLL) is the most common haematological malignancy in the U.S. The course of the disease has been shown to be negatively impacted by increased levels of BCL2. Strategies to downregulate BCL2 and shift the balance towards cellular demise are actively being explored. Therefore, we examined whether the investigational agent MLN2238 could inhibit the proteasomal machinery and induce CLL cell death while also downregulating BCL2. MLN2238-induced cell death was studied in peripheral blood mononuclear cells from 28 CLL patients. MLN2238 produced a dose-dependent reduction in BCL2 and CLL cell viability with maximum cell death observed at a 50 nmol/l concentration by 48 h. Annexin-V staining, PARP1 and caspase-3 cleavage along with an increase in mitochondrial membrane permeability were noted after cells were treated with MLN2238; however, apoptosis was only partially blocked by the pan-caspase inhibitor z-VAD.fmk. Furthermore, we observed enhanced anti-CLL effects in tumour cells treated with either a combination of MLN2238 and the BH3 mimetic AT-101 or MLN2238 and fludarabine. Together, our data suggest the potential for proteasome inhibitor based therapy in CLL and the rationale design of drug combination strategies based on CLL biology.

Salivary proteins of plant-feeding hemipteroids - implication in phytophagy.

Many hemipteroids are major pests and vectors of microbial pathogens, infecting crops. Saliva of the hemipteroids is critical in enabling them to be voracious feeders on plants, including the economically important ones. A plethora of hemipteroid salivary enzymes is known to inflict stress in plants, either by degrading the plant tissue or by affecting their normal metabolism. Hemipteroids utilize one of the following three strategies of feeding behaviour: salivary sheath feeding, osmotic-pump feeding and cell-rupture feeding. The last strategy also includes several different tactics such as lacerate-and-flush, lacerate-and-sip and macerate-and-flush. Understanding hemipteroid feeding mechanisms is critical, since feeding behaviour directs salivary composition. Saliva of the Heteroptera that are specialized as fruit and seed feeders, includes cell-degrading enzymes, auchenorrhynchan salivary composition also predominantly consists of cell-degrading enzymes such as amylase and protease, whereas that of the Sternorhyncha includes a variety of allelochemical-detoxifying enzymes. Little is known about the salivary composition of the Thysanoptera. Cell-degrading proteins such as amylase, pectinase, cellulase and pectinesterase enable stylet entry into the plant tissue. In contrast, enzymes such as glutathione peroxidase, laccase and trehalase detoxify plant chemicals, enabling the circumvention of plant-defence mechanisms. Salivary enzymes such as M1-zinc metalloprotease and CLIP-domain serine protease as in Acyrthosiphon pisum (Aphididae), and non-enzymatic proteins such as apolipophorin, ficolin-3-like protein and 'lava-lamp' protein as in Diuraphis noxia (Aphididae) have the capacity to alter host-plant-defence mechanisms. A majority of the hemipteroids feed on phloem, hence Ca++-binding proteins such as C002 protein, calreticulin-like isoform 1 and calmodulin (critical for preventing sieve-plate occlusion) are increasingly being recognized in hemipteroid-plant interactions. Determination of a staggering variety of proteins shows the complexity of hemipteroid saliva: effector proteins localized in hemipteran saliva suggest a similarity to the physiology of pathogen-plant interactions.

The importance of ultrasound in staging and gaining a pathological diagnosis in patients with lung cancer--a two year single centre experience.

BACKGROUND: Initial studies on the use of ultrasound in the detection and sampling of supraclavicular lymph nodes in patients with suspected lung cancer show this to be a promising technique, giving both a cytological diagnosis and pathological N3 (pN3) stage. Leicester published its initial experience in 2005 and the aim of this study was to establish if this had been embedded into the diagnostic pathway, and further to examine the use of ultrasound in diagnosing and staging lung cancer by imaging other areas including pleural effusions, chest wall, bone and liver lesions. METHODS: All patients diagnosed with lung cancer, registered on the Leicester lung cancer database over a two year period between January 2007 and December 2008, had their imaging and pathology retrospectively reviewed; 996 primary lung cancer patients were identified (n=996). Of these, 318 patients underwent an ultrasound examination (n=318), consisting of ultrasound of the neck, pleural cavity, and metastatic lesions potentially amenable to ultrasound guided aspiration/biopsy. RESULTS: The overall malignant yield was 45% of patients scanned (95% CI 39.5% to 50.4%) and 81.3% of patients sampled (95% CI 75.5% to 87%). Of the 996 patients, 14.4% (n=143) had a positive ultrasound guided cytological diagnosis (95% CI 12.2% to 16.5%). Of all the pathological diagnoses (n=765), 18.7% were ultrasound guided (95% CI 15.9% to 21.5%). In particular, 32.2% of patients with CT detected neck or mediastinal nodes had a diagnosis and stage achieved by neck ultrasound. CONCLUSION: The use of ultrasound gives a rapid and less invasive method of diagnosing and staging lung cancer and has become embedded into the diagnostic pathway. We advocate its increased use and availability in patients with lung cancer.

RNA editing enzyme APOBEC3A promotes pro-inflammatory M1 macrophage polarization.

Pro-inflammatory M1 macrophage polarization is associated with microbicidal and antitumor responses. We recently described APOBEC3A-mediated cytosine-to-uracil (C > U) RNA editing during M1 polarization. However, the functional significance of this editing is unknown. Here we find that APOBEC3A-mediated cellular RNA editing can also be induced by influenza or Maraba virus infections in normal human macrophages, and by interferons in tumor-associated macrophages. Gene knockdown and RNA_Seq analyses show that APOBEC3A mediates C>U RNA editing of 209 exonic/UTR sites in 203 genes during M1 polarization. The highest level of nonsynonymous RNA editing alters a highly-conserved amino acid in THOC5, which encodes a nuclear mRNA export protein implicated in M-CSF-driven macrophage differentiation. Knockdown of APOBEC3A reduces IL6, IL23A and IL12B gene expression, CD86 surface protein expression, and TNF-α, IL-1ß and IL-6 cytokine secretion, and increases glycolysis. These results show a key role of APOBEC3A cytidine deaminase in transcriptomic and functional polarization of M1 macrophages.

Radiographic and functional evaluation of low profile dorsal versus volar plating for distal radius fractures.

INTRODUCTION: Fracture of the distal radius is a common clinical problem. Complex fracture requires open reduction and stabilization with plating to restore anatomy. Dorsal plating has advantages of buttressing the fracture better but often complicated with tendon problems as per literature. The rate of complications however, was not compared between the low-profile dorsal and the volar plates. METHODOLOGY: This was a retrospective study on seventy one patients with dorsally angulated or displaced distal radius fractures, who underwent fixation of fractures with either dorsal or volar locking plate from Jan - Nov 2012. Preoperative radiographs were classified based on Universal and Fernandez classification. Postoperative radiographs were assessed for anatomical restoration of Radial length, radial inclination and volar tilt. Tendon and nerve related complications were assessed and functional evaluation was performed on the basis of PRWE (Patient related wrist evaluation) score. RESULTS: Both groups were matched for their demographic profile and fracture types (p 0.033). Dorsal plating group had 89% excellent/good restoration and fair in 11%. Volar group had 96% excellent/good restoration and fair in 4%. Statistical analysis was performed with unpaired t test for radiographic parameters. Three patients had tendon related complications in dorsal plating group; two patients in volar group had nerve related complications. Functional outcome with PRWE was comparable between two groups. CONCLUSION: Results with low profile dorsal plating were comparable to volar plating. Therefore dorsal plating can be used as an alternative method when dorsal buttressing of comminuted fracture is required, especially with concomitant osteoporosis.

Current scenario of spurious and substandard medicines in India: a systematic review.

Globally, every country is the victim of substandard or spurious drugs, which result in life threatening issues, financial loss of consumer and manufacturer and loss in trust on health system. The aim of this enumerative review was to probe the extent on poor quality drugs with their consequences on public health and the preventive measures taken by the Indian pharmaceutical regulatory system. Government and non-government studies, literature and news were gathered from journals and authentic websites. All data from 2000 to 2013 were compiled and interpreted to reveal the real story of poor quality drugs in India. For minimizing spurious/falsely-labelled/falsified/counterfeit drugs or not of standard quality drugs, there is urgent requirement of more stringent regulation and legal action against the problem. However, India has taken some preventive steps in the country to fight against the poor quality drugs for protecting and promoting the public health.

Technetium 99m-labelled-sucralphate isotope scanning in the detection of peptic ulceration.

An isotope scan using sucralphate labelled with technetium-99m has been compared with barium meal in the detection of peptic ulceration in 18 patients with endoscopically proven upper gastrointestinal lesions and eight controls. Gastric (75%) and duodenal (67%) ulcers were detected with identical frequency by isotope scan and barium meal but both were inferior to endoscopy. Structural, non-ulcerated lesions, were not identified by isotope scanning. The scan has potential value in duodenal ulcer disease, the serial assessment of ulcer healing and in paediatric gastroenterology.

Enhanced 99Tc(m)-MIBI SPECT detection of hibernating myocardium following the use of sub-lingual nitroglycerine.

The purpose of this study was to assess the feasibility of detecting hibernating myocardium using 99Tc(m)-MIBI SPECT imaging following the use of sublingual glyceryl trinitrate (GTN). Thirty-eight subjects were studied, five asymptomatic and 33 with a history of myocardial ischaemia or myocardial infarction (MI). The study was conducted on two separate days, both at rest, one under controlled basal conditions and the other 20 min following the sublingual administration of GTN. A total of 190 cardiac segments were qualitatively analysed and 72 segments with fixed defects were compared to the study following the use of GTN for evidence of improved radionuclide uptake. Following GTN, 73.6% of fixed segment defects showed no change in uptake, 5.5% showed probable improvement whilst in 20.8% a definite enhanced uptake was demonstrated. In 3.7% of segments, worsening of radionuclide uptake was observed.

Low seroconversion with hepatitis B vaccination in peritoneal dialysis patients.

OBJECTIVE: To compare seroconversion using hepatitis B vaccine between hemodialysis (HD) and peritoneal dialysis (PD) patients. DESIGN: Data on PD patients vaccinated were collected retrospectively for the period 1992 to 1995. The data on HD patients were collected prospectively from 1991 to 1994. SETTING: A university outpatient dialysis center. PARTICIPANTS: All adult patients who received all four doses of hepatitis B vaccine while on dialysis were included (47 PD and 50 HD patients). INTERVENTION: Recombinant hepatitis B vaccine (Engerix), 40 micrograms IM was administered at 0, 1, 2, and 6 months. MAIN OUTCOME MEASURE: Seroconversion was measured after completion of the vaccination series. RESULTS: 74% of the HD patients seroconverted compared to 53% of PD patients (p = 0.03). Older, heavier patients compared to all the other patients had a lower seroconversion rate in both the HD patients (55% vs. 78 %) and PD patients (38% vs. 59%) (p = 0.03). CONCLUSION: The seroconversion rate to recombinant hepatitis B vaccine is lower in patients on PD than on HD for unclear reasons. Further studies are required to determine the etiology of this difference.

Skull base involvement by a nasopharyngeal carcinoma shown by Tc-99m MDP SPECT but not by computed tomography.

PURPOSE: Plain radiographs, computed tomography (CT), and more recently magnetic resonance imaging (MRI) are used routinely to stage carcinoma of the nasopharynx. Tc-99m methylene diphosphonate (MDP) SPECT is seldom used for local staging of the disease. MATERIALS AND METHODS: Plain radiographs and CT were used to stage squamous carcinoma of the nasopharynx in a 50-year-old man with a left XII nerve palsy. RESULTS: Findings of the plain radiographs were normal, whereas the CT scan revealed a nonhomogenous hyperdense mass in the nasopharynx but intact underlying bone. Given the symptoms, a Tc-99m MDP planar scan was ordered and showed no enhanced uptake, but SPECT images obtained at the same time revealed markedly increased focal radiotracer uptake in the region of the tumor, indicating osseous involvement. CONCLUSION: Possible bony invasion with a nasopharyngeal carcinoma may be better shown with MDP SPECT than with planar isotope bone scans, plain radiographs, or CT.

Prescribing practices of consultants at Karachi, Pakistan.

OBJECTIVE: To audit the prescribing practices of consultants. DESIGN: A randomized sample survey of 354 consultants prescription for analysis of some basic parameters as recommended by World Health Organization (WHO). MAIN OUTCOME MEASURES: To provide feedback to the prescribers for promoting rational drug therapy. RESULTS: The average number of drugs for prescription were found to be 4.51 and the average daily cost of prescribed drugs was Rs: 133.41. The antimicrobials, vitamins/minerals and injections were over-prescribed. About half of the prescribed drugs were from the National Essential Drugs List of Pakistan (NEDLP) and only a little over 12% were prescribed by their generic names. CONCLUSIONS: The prescribing practices of the consultants in Karachi are rather liberal and not rational. Overuse of antimicrobials and injections have been observed and there is general tendency of indulging in polypharmacy requiring continued medical education.

Detection of early xerophthalmia by impression cytology and rose Bengal staining--a comparative study.

Twenty two cases were studied for early detection of Xerophthalmia by impression cytology and Rose Bengal staining. Patients were below 6 years of age and admitted in the paediatric ward of Dhaka Medical College Hospital. Assessment were done by conjunctival impression cytology and staining with 1% Rose Bengal dye. Estimation of serum retinol level was done by HPLC to correlate conjunctival findings with biochemical status. The mean age of the subjects were 3.15 +/- 1.75. Rose Bengal staining was positive in 40% cases and impression cytology was in 60% cases. The mean serum retinol level of the diseased children were 10.5 +/- 3. ug/dl. The sensitivity of Rose Bengal staining was 53.3% and that of impression cytology was 80% in detecting vitamin A deficiency when compared with serum retinol level. The specificity of Rose Bengal staining was 40% and that of impression cytology was 100%. Sensitivity of impression cytology was 100% in patient with retinol level 10 ug/dl.