Enhanced morphological transformation of human lung epithelial cells by continuous exposure to cellulose nanocrystals.

Cellulose nanocrystals (CNC), also known as nanowhiskers, have recently gained much attention due to their biodegradable nature, advantageous chemical and mechanical properties, economic value and renewability thus making them attractive for a wide range of applications. However, before these materials can be considered for potential uses, investigation of their toxicity is prudent. Although CNC exposures are associated with pulmonary inflammation and damage as well as oxidative stress responses and genotoxicity in vivo, studies evaluating cell transformation or tumorigenic potential of CNC's were not previously conducted. In this study, we aimed to assess the neoplastic-like transformation potential of two forms of CNC derived from wood (powder and gel) in human pulmonary epithelial cells (BEAS-2B) in comparison to fibrous tremolite (TF), known to induce lung cancer. Short-term exposure to CNC or TF induced intracellular ROS increase and DNA damage while long-term exposure resulted in neoplastic-like transformation demonstrated by increased cell proliferation, anchorage-independent growth, migration and invasion. The increased proliferative responses were also in-agreement with observed levels of pro-inflammatory cytokines. Based on the hierarchical clustering analysis (HCA) of the inflammatory cytokine responses, CNC powder was segregated from the control and CNC-gel samples. This suggests that CNC may have the ability to influence neoplastic-like transformation events in pulmonary epithelial cells and that such effects are dependent on the type/form of CNC. Further studies focusing on determining and understanding molecular mechanisms underlying potential CNC cell transformation events and their likelihood to induce tumorigenic effects in vivo are highly warranted.

Global Collaborations for Cervical Cancer: Can the East-West Alliance Facilitate Treatment for all?

Despite the advances in the primary prevention of cervical cancer, there is an absolute increase in the incidence of cervical cancer as a result of an increase in world population. A vast majority of patients in low and low-middle income countries continue to present at a locally advanced stage, necessitating treatment with chemoradiation and brachytherapy. There is a dearth of equipment and trained professionals for the treatment of cervical cancer, especially in low and low-middle income countries. There is an urgent need to improve treatment availability and develop better treatments. Worldwide trends, however, reveal a low number of therapeutic and innovative research trials in cervical cancer. The present article elucidates the existing challenges and provides solutions to improve outcomes. The proposed strategies hinge on strengthening collaborations for global advocacy.

Postgraduate Global Health Competency Profile for Radiation Oncology.

AIMS: To identify core competencies for postgraduate radiation oncology trainees in global health and cancer that may inform revisions across radiation oncology residency specialty training curricula. MATERIALS AND METHODS: A review of the literature was conducted to identify all potential global health competency items. An international two-phase Delphi process was conducted with experts in oncology. In phase 1, all experts scored, on a nine-point Likert scale, the degree to which they agreed an item should be included in the competency profile. Items with a mean score ≥7 were included, those scoring 4-6 were reviewed in phase 2, and items scored ≤3 were excluded. In phase 2, items were discussed and subsequently ranked for inclusion or exclusion in the competency profile. Items with >75% voting for inclusion were included in the final competency profile. RESULTS: In total, 142 potential global health competency items were identified. Sixty-one items were removed as redundant or irrelevant, leaving 81 items for the Delphi process. Eighteen specialists were invited to participate, with 10 (56%) agreeing to participate in phase 1 of the Delphi process. Participants represented 10 centres in seven countries. Of the 81 items ranked in phase 1, 72 items (89%) had a mean score ≥7 and were automatically included in the final competency profile. The remaining nine items received a score of 4-6 and were discussed in phase 2 of the Delphi process, of which three received <75% agreement for inclusion and were excluded. The result was a final list of 78 enabling competency items. CONCLUSIONS: The radiation oncology global health competency profile represents an international consensus on the items that can inform radiation oncology training requirements.

Mobilising Expertise and Resources to Close the Radiotherapy Gap in Cancer Care.

Closing the gap in cancer care within low- and middle-income countries and in indigenous and geographically isolated populations in high-income countries requires investment and innovation. This is particularly true for radiotherapy, for which the global disparity is one of the largest in healthcare today. New models and paradigms and non-traditional collaborations have been proposed to improve global equity in cancer control. We describe recent initiatives from within the radiation oncology community to increase access to treatment, build the low- and middle-income countries' radiation oncology workforce, mobilise more professionals from within high-income countries and raise awareness of the global need for equitable cancer care.

Balancing Equity and Advancement: The Role of Health Technology Assessment in Radiotherapy Resource Allocation.

Radiotherapy is an essential modality for effective cancer control, yet enormous inequalities in access in low- and middle-income countries (LMICs) have created one of the largest global technology gaps in medicine today. The Global Task Force on Radiotherapy for Cancer Control quantified this gap and showed that over half of patients worldwide do not have access to treatment. Governments, policy makers and the global health community have ignored this crisis due to the complexity of radiotherapy technology and its seemingly high upfront costs. However, understanding the cost of treatment in the context of a dramatic clinical benefit could help to demonstrate the feasibility of radiotherapy in diverse income settings. When there are scarce resources, such analysis is essential in order to set priorities and provide high-value interventions to large populations. Here we explore the current status of economic evaluation tools in LMICs and some of the barriers to their use. We describe how the concepts of health technology assessment, value-based care and investment frameworks can be applied to the global crisis of radiotherapy availability to guide appropriate capacity building and resource utilisation. The development of local expertise in these health economic tools can be a powerful level to improve cancer care in LMICs and to build universal global access to radiotherapy.

The Brain Metastases Symptom Checklist as a novel tool for symptom measurement in patients with brain metastases undergoing whole-brain radiotherapy.

PURPOSE: We evaluated the feasibility, reliability, and validity of the Brain Metastases Symptom Checklist (bmsc), a novel self-report measure of common symptoms experienced by patients with brain metastases. METHODS: Patients with first-presentation symptomatic brain metastases (n = 137) referred for whole-brain radiotherapy (wbrt) completed the bmsc at time points before and after treatment. Their caregivers (n = 48) provided proxy ratings twice on the day of consultation to assess reliability, and at week 4 after wbrt to assess responsiveness to change. Correlations with 4 other validated assessment tools were evaluated. RESULTS: The symptoms reported on the bmsc were largely mild to moderate, with tiredness (71%) and difficulties with balance (61%) reported most commonly at baseline. Test-retest reliability for individual symptoms had a median intraclass correlation of 0.59 (range: 0.23-0.85). Caregiver proxy and patient responses had a median intraclass correlation of 0.52. Correlation of absolute scores on the bmsc and other symptom assessment tools was low, but consistency in the direction of symptom change was observed. At week 4, change in symptoms was variable, with improvements in weight gain and sleep of 42% and 41% respectively, and worsening of tiredness and drowsiness of 62% and 59% respectively. CONCLUSIONS: The bmsc captures a wide range of symptoms experienced by patients with brain metastases, and it is sensitive to change. It demonstrated adequate test-retest reliability and face validity in terms of its responsiveness to change. Future research is needed to determine whether modifications to the bmsc itself or correlation with more symptom-specific measures will enhance validity.

Expression of human amyloid precursor protein in Drosophila melanogaster nerve cells causes a decrease in presynaptic gene mRNA levels.

Amyloid precursor protein (APP) is a key player in Alzheimer's disease. The proteolytic cleavage of APP results in various short peptide fragments including the toxic amyloid-beta peptide, which is a main component of senile plaques. However, the functions of APP and its processed fragments are not yet well understood. Here, using real-time polymerase chain reaction, we demonstrate that exogenous expression of APP, its mutant form APP-Swedish, or two truncated forms in Drosophila melanogaster causes a significant (P ≤ 0.05) drop in the mRNA levels of the presynaptic proteins synaptotagmin-1 and neuronal synaptobrevin. The results obtained from this study suggest a potential role of APP or its fragments in the regulation of synaptic gene transcription.

Pregnancy outcomes in women with gestational diabetes treated with metformin or insulin: a case-control study.

AIMS: To compare maternal and neonatal outcomes in women with gestational diabetes mellitus (GDM) treated with either metformin or insulin. METHODS: One hundred and twenty-seven women with GDM not adequately controlled by dietary measures received metformin 500 mg twice daily initially. The dose was titrated to achieve target blood glucose values. Pregnancy outcomes in the 100 women who remained exclusively on metformin were compared with 100 women with GDM treated with insulin matched for age, weight and ethnicity. RESULTS: There were no significant differences in baseline maternal risk factors. Women treated with insulin had significantly greater mean (sem) weight gain from enrolment to term (2.72 +/- 0.4 vs. 0.94 +/- 0.3 kg; P < 0.001). There was no difference between the metformin and insulin groups, respectively, comparing gestational hypertension (6 vs. 7%, P = 0.9), pre-eclampsia (9 vs. 2%, P = 0.06) induction of labour (26 vs. 24%, P = 0.87) or rate of Caesarean section (48 vs. 52%, P = 0.67). No perinatal loss occurred in either group. Neonatal morbidity was improved in the metformin group; prematurity (0 vs. 10%, P < 0.01), neonatal jaundice (8 vs. 30%, P < 0.01) and admission to neonatal unit (6 vs. 19%, P < 0.01). The incidence of macrosomia (birthweight centile > 90) was not significantly different [metformin (14%) vs. insulin (25%); P = 0.07]. CONCLUSIONS: Women with GDM treated with metformin and with similar baseline risk factors for adverse pregnancy outcomes had less weight gain and improved neonatal outcomes compared with those treated with insulin. Diabet. Med. 26, 798-802 (2009).

[Studying pathogenesis of Alzheimer's disease in a Drosophila melanogaster model: human APP overexpression in the brain of transgenic flies leads to deficit of the synaptic protein synaptotagmin].

Alzheimer's disease (AD) is a progressive neurodegenerative disease whose main pathomorphological sign is synapse degeneration in the cortex and hippocampus. Abnormal synaptogenesis precedes amyloidosis and neurodegeneration and correlates with memory impairment during the early clinical phase. Mutations in the amyloid precursor protein (APP) gene cause familial AD and enhance the secretion of amyloid-beta-protein (Abeta). However, it remains unclear in what way APP and Abeta are involved in synaptic disorder in the absence of visible amyloid structures. In this study, the role of the human APP gene in synaptogenesis in transgenic lines of Drosophila melanogaster whose nerve cells express the human APP695 isoform, truncated APPs, and the presynaptic marker synaptotagmin driving the sequence of the green fluorescent protein. The expression of APP and its truncated forms caused a decrease in the synaptotagmin content of antennal lobes and mushroom lobes of the D. melanogaster brain, as well as neurodegeneration that progressed with age. The results suggest that that abnormal synaptogenesis and neurodegeneration occur in the Drosophila brain in the absence of Abeta. It is assumed that impaired cellular functions of APP and secretion of Abeta independently contribute to the pathogenesis of AD.

[Changes in physicochemical properties of proteins, caused by modification with alkylhydroxybenzenes].

Kinetic characteristics of model enzymes and physicochemical properties of globular proteins modified by chemical analogues of low-molecular-weight microbial autoregulators (alkylhydroxybenzenes, AHBs) have been studied. C7 and C12 AHB homologues were used, differing in the length of the alkyl radical and the capacity for weak physicochemical interactions. Both homologues affected the degree of protein swelling, viscosity, and the degree of hydrophobicity. The effects depended on the structure of AHBs, their concentration, and pH of the solution, which likely reflects changes in the charge of the protein globule and its solvate cover. Variations of hydrophobicity indices of AHB-modified enzymes (trypsin and lysozyme) were coupled to changes in the catalytic activity. The values of K(M), measured for the enzymes within both AHB complexes, did not change, whereas V(max) increased (in the case of C7 complexes) or decreased (C12 complexes). Possible molecular mechanisms of changes in the physicochemical and catalytic parameters of enzymatically active proteins, induced by modification with structurally distinct AHBs, are described, with emphasis on targeted regulation of functional activity.

[Transurethral endoscopic pyelolithotripsy in combined treatment of complex forms of nephrolithiasis].

The article presents results of retrograde endoscopic pyelolithotripsy (REP) in 75 patients with large and coral nephroliths. Retrograde contact pyelolithotripsy was made with application of Swiss Lithoclast unit, semirigid ureteroscopes R. Wolf with a cone 8-9 Ch and working channel 5.3 Ch. This enables maximal removal of the destroyed concrement by hydraulic litholapaxia with consequent insertion of the internal stent. Extracorporeal lithotripsy with low-energy shock-wave impulses was performed on demand (1-3 sessions). The stent was removed in 3-5 days in cases of complete one-stage sanation of the kidney and in 2-3 weeks in the presence of residual fragments of the nephrolith. One-stage complete sanation in coral nephrolithiasis (K1-K2) up to 5 cm3 and large concrements was achieved in 69.3% patients. Three months later residual fragments of the nephroliths were observed in 8% patients, 6 months later--in 5.3%. One year after treatment a complete sanation was achieved in 96%. Complications of the method consisted in exacerbation of chronic pyelonephritis observed in 12% cases. The control examination revealed improvement of renal function early and late after surgery. As shown by 6-year follow-up, recurrent nephroliths occurred in 6% patients. Endoscopic retrograde pyelolithotripsy can be used as a method of choice in the treatment of compound forms of nephrolithiasis in intrarenal type of the pelvis, the absence of marked extension of the caliceal-pelvic system of the kidney, in other stones resistant to extracorporeal lithotripsy.

Expression of the chimeric IgE gene in cell culture and in various mouse tissues.

Transient expression of recombinant gene constructs is now more widely used in gene therapy as well as in DNA vaccination. In this study, the ability of one and the same genetic construct to drive gene expression both in cell culture and in tissues of the whole organism was demonstrated. Chinese hamster ovarian cells (CHO) were transfected in vitro with plasmids bearing the genes for chimeric IgE (mouse/human) antibodies under control of the human cytomegalovirus (hCMV) promoter. Secretion of recombinant IgE antibodies by transfected cells reached 60% of the intracellular concentration of antibodies. The same gene constructs were introduced into various mouse tissues using ballistic transfection in vivo. The IgE content in blood after transfection of cartilage was found to be several times lower than after transfection of the liver, spleen, or foot pad. At the same time, the content of antibodies to the xenogenous determinants of IgE was essentially independent of the tissue type. These data can be employed in selecting conditions for genetic immunization and gene therapy.

[Blood transfusion support in rendering medical care in extreme conditions]. / Problema transfuziologicheskogo obespecheniia pri okazanii meditsinskoi pomoshchi v ékstremal'nykh usloviiakh.

About 20% of accident and disaster victims need the hemotransfusion therapy. At present the method of erythrocyte long-term storage in the blood banks with glycerin high concentration is the most acceptable one. There are special medical teams (SMT) in the districts. They include the transfusiologic group with appropriate reserve of hemotransfusion materials. Most of them must be stored as untouchable reserve in the institutions of blood service and large hospitals on which the SMTs are based.

Is the defect in pro-hormone processing in Type 2 diabetes mellitus restricted to the beta cell?

AIMS: To investigate whether the defect in pro-hormone processing in people with Type 2 diabetes mellitus is restricted to the pancreatic beta cell or whether there is evidence of a more generalized abnormality. METHODS: Ten Indian subcontinent Asian women with diet-controlled Type 2 diabetes were compared with a control group of nine non-diabetic Asian women who were matched for body mass index. All subjects underwent a standard 75 g oral glucose tolerance test during which plasma glucose, insulin, proinsulin and 32,33 split proinsulin were measured. Subjects in both groups then underwent an intravenous corticotrophin-releasing hormone (CRH) test with 100 microg human CRH. Plasma cortisol, adrenocorticotrophic hormone (ACTH) and ACTH precursors were measured. RESULTS: Basal levels of insulin were lower in diabetic subjects (32.5 (12.5-52) pmol/l) than in the control group (42.2 (21.4-63.0) pmol/l); normalized geometric mean (95% confidence interval), P<0.05; as were the levels at 30 min (29.5 (3.7-55.3) pmol/l) and (34.4 (10.7-58.2) pmol/l), P<0.05. The levels at 60 min were (26.7 (6.4-47.0) pmol/l) in subjects with diabetes and (13.6 (-11.3-38.5) pmol/l) in controls, P<0.05 and at 90 min these were (43.4 (19.4-67.4) pmol/l) and (19.3 (-4.6-43.3) pmol/l), P<0.05, respectively, after the 75-g oral glucose load. The acute insulin response was markedly reduced in the subjects with diabetes (1.8 (0.60-3.1) pmol insulin/ mmol glucose), compared with the control group (31.4 (8.0-54) pmol/l insulin/mmol glucose), P<0.005. Intact proinsulin was much higher in the diabetic subjects (23.9 (10.1-37) pmol/1) than in the control group (7.2 (3.9-10) pmol/1), P<0.002, as was the percentage of proinsulin-like molecules (28 (14-42) and 12 (8-17)%), respectively, P<0.01. There were no differences between basal or stimulated levels of ACTH precursors, ACTH or cortisol between the diabetic subjects and the controls. CONCLUSIONS: The defect in insulin secretion in Indian subcontinent Asian women with Type 2 diabetes is similar to that described in other populations with an increased prevalence of Type 2 diabetes. The absence of any defect in the processing of cortisol precursors in the women with Type 2 diabetes suggests that they do not have a generalized defect of hormone processing.

The quantitative characteristics of efficiency of ballistic transfection of chimeric antibody genes.

A quantitative approach was applied to the study of in vivo expression of foreign genes introduced into mice by ballistic transfection. Because in some cases one must take into account both the level of synthesized protein and that of antibodies to it, we derived the equation which allows to calculate the exact quantity of both proteins. This formula was applied to in vivo expression of a chimeric (human/mice) immunoglobulin E gene. The immunochemical analysis using this equation showed that the Ig concentration succeeded 4, 6, 12 IU/ml and undetectable level, respectively, upon transfection in mouse liver, spleen, foot pad and ear cartilage.

Reduced bone mineral density after surgical treatment for obesity.

OBJECTIVE: To investigate whether osteoporosis occurs after surgical treatment for obesity. DESIGN: A cross-sectional study of five groups of subjects who had undergone surgical treatment for obesity: five pre-menopausal women; 13 post-menopausal women; seven post-menopausal women taking oestrogen replacement (HRT); five men; and six women who had undergone surgical reversal (mean time 7 y). SUBJECTS: Thirty-six Caucasian subjects who had undergone jejunoileal or pancreaticobiliary bypass surgery at St George's Hospital between 1971 and 1992. Their mean age was 50.8 y (range 32-69 y) and the median time since the operation was 14.8y (range 4-23 y). MEASUREMENTS: A clinical questionnaire was used to exclude possible factors, which might influence bone mineral density. A single blood sample was collected for measurement of calcium, phosphate, alkaline phosphatase, albumin, magnesium, zinc, creatinine, thyroxine, 25-hydroxy-vitamin D, sex steroids, gonadotrophins and IGF-1 and 24 h urine calcium excretion was measured. Bone mineral density (BMD) was measured in the lumbar (L2-L4) spine (LS) and femoral neck (FN) by dual energy X-ray absorptiometry (DEXA). RESULTS: There was no difference in serum calcium, alkaline phosphatase, IGF-1, 25-hydroxy-vitamin D (25-OH vitamin D), magnesium or zinc concentrations between the five groups. The LS-BMD T score was lower (P < 0.05) in male subjects ( -2.08 +/- 1.04 mean 1.0 +/- s.d) and post-menopausal women not taking HRT ( -1.21 +/- 1.33) compared to the surgically reversed group (0.87 +/- 2.36). The male group was most severely affected, despite normal serum testosterone concentrations. Two of the five men studied, had a LS-BMD T score < -2.5 and two had a LS-BMD T score between -1.0 and -2.5. In contrast, six of the seven post-menopausal women on HRT had an LS BMD T score > - 1.0. There was no difference in the FN-BMD between the five groups. The presence of low BMD was not related to age, duration of bypass, or degree of postoperative weight loss. Iliac crest bone biopsies in three subjects with low BMD, confirmed the presence of osteoporosis. CONCLUSIONS: Reduced bone mineral density is a complication of jejunoileal bypass surgery.

Polycystic ovaries and associated metabolic abnormalities in Indian subcontinent Asian women.

OBJECTIVES: To determine the prevalence of polycystic ovaries (PCO) in Asian women living in England who are of Indian subcontinent origin or ancestry and to investigate the relationship between the presence of PCO and/or non-insulin dependent diabetes mellitus (NIDDM) and insulin sensitivity and other metabolic parameters. DESIGN: A random sample of Indian subcontinent Asian women was obtained from the lists of local General Practitioners and a translating service. These women were invited to attend for a medical history questionnaire, examination, venous blood sample for hormonal assessment and transvaginal ovarian ultrasonography. Groups of women without PCO or NIDDM, with NIDDM but not PCO, with PCO but not NIDDM and with both NIDDM and PCO were drawn at random from this population and from Indian subcontinent Asian women attending the Diabetes Unit. They underwent further studies, including measurement of insulin sensitivity using a short intravenous insulin tolerance test. SUBJECTS: 212 Indian subcontinent Asian women aged 18-40 took part in the initial study. Insulin sensitivity was measured in 13 women without PCO or NIDDM, 13 women with NIDDM but not PCO, 15 women with PCO but not NIDDM and 12 women with both NIDDM and PCO. MEASUREMENTS: The main outcome measures were prevalence of polycystic ovaries, clinical features of hyperandrogenism, fertility, blood pressure, serum gonadotrophins, testosterone and sex hormone binding globulin, fasting blood lipids, glucose and insulin, and insulin sensitivity. RESULTS: The prevalence of PCO in Indian subcontinent Asian women was 52% (110/212). There were significant associations between PCO and menstrual irregularity; infertility; the Ferriman and Gallwey score for body hair distribution; the presence of acanthosis nigricans and the fasting blood glucose concentration. There were no differences between women with PCO and those with normal ovarian morphology with respect to systolic and diastolic blood pressure, fasting total, HDL and LDL cholesterol and triglyceride concentrations. The subgroup of women without PCO or NIDDM had the highest insulin sensitivity (189.1 +/- 46.4 mumol glucose/l/min, mean +/- SD) and the women with both PCO and NIDDM had the lowest insulin sensitivity (80.5 +/- 30.9 mumol glucose/l/min). There was no significant difference in insulin sensitivity between those with PCO but not NIDDM (125.0 +/- 59.5 mumol glucose/l/min) and those with NIDDM but not PCO (120.8 + 38.0 mumol glucose/l/min). The effects of NIDDM and PCO on insulin sensitivity were independent; the effect of PCO on insulin sensitivity was -60 mumol glucose/l/min (95% confidence interval -100 to -21, P = 0.004) and the effect of NIDDM was -68 mumol glucose/l/min (95% confidence interval -105 to -31, P < 0.001). There were no significant relationships between insulin sensitivity and fasting plasma insulin, systolic or diastolic blood pressure, fasting serum cholesterol or triglyceride. CONCLUSIONS: The prevalence of polycystic ovaries in Indian subcontinent Asian women is very high and it has significant clinical associations. Polycystic ovaries and non-insulin dependent diabetes mellitus are associated with similar degrees of reduced insulin sensitivity in this population. Their effects are independent suggesting that these changes in insulin sensitivity involve different mechanisms. Polycystic ovaries unlike non-insulin dependent diabetes mellitus, are not associated with a defect in the secretion of insulin.