Cell Cycle Analysis of ER Stress and Autophagy.

Autophagy and ER stress are most often studied employing a Western blotting approach to the measurement of autophagy by LC3B upregulation and the ER stress sensor signaling proteins PERK (protein kinase R-like endoplasmic reticulum kinase), IRE1, and ATF6 which initiate protein refolding and elongation of the ER until ER homeostasis is returned. If the misfolding of proteins is increased, then ER stress is maintained, and microautophagy of the ER or specifically reticulophagy occurs. However, LC3B, PERK, protein misfolding, and changes in ER mass (reticulophagy) can also be measured in a cell cycle-dependent manner by flow cytometry and the use of antibodies, protein misfolding, and ER tracking fluorescent probes.

Comprehensive Analysis of Adverse Events Associated with Gastric Peroral Endoscopic Myotomy: An International Multicenter Study.

BACKGROUND: Gastric peroral endoscopic myotomy (G-POEM) has emerged as an effective management approach for patients with refractory gastroparesis. This study aims to comprehensively study the safety of G-POEM and describe the predictive factors of adverse events (AEs) occurrence. METHODS: This study is a retrospective study involving 13 tertiary care centers (7 USA, 1 South America, 4 Europe, and 1 Asia). Patients who underwent G-POEM for refractory gastroparesis were included. Cases were identified by the occurrence of AEs. For each case, two controls were randomly selected and matched for age (± 10 years), gender, and etiology of gastroparesis. RESULTS: A total of 216 patients underwent G-POEM for gastroparesis. Overall, 31 (14%) AEs were encountered [mild 24 (77%), moderate 5 (16%), and severe 2 (6%)] during the duration of the study. The most common AE was abdominal pain (n = 16), followed by mucosotomy (n = 5) and capnoperitoneum (n = 4), and AEs were most commonly identified within the first 48-h post-procedure 18 (58%). The risk of adverse event occurrence was significantly higher for endoscopists with experience of < 20 G-POEM procedures (OR 3.03 [1.03-8.94], p < 0.05). CONCLUSION: G-POEM seems to be a safe intervention for refractory gastroparesis. AEs are most commonly mild and managed conservatively. Longitudinal mucosal incision, use of hook knife, use of clips for mucosal closure and endoscopist's experience with > 20 G-POEM procedures is significantly associated with decreased incidence of AEs.

Initial experience of CT-guided pulsed radiofrequency ablation of the pudendal nerve for chronic recalcitrant pelvic pain.

AIM: To evaluate initial experience with computed tomography (CT)-guided pulsed radiofrequency ablation (pRFA) of the pudendal nerve in cases of recalcitrant neuropathic pelvic pain. Endpoints include technical feasibility, safety, and efficacy of therapy. MATERIALS AND METHODS: Ten patients who underwent pRFA ablation for neuropathic pudendal nerve pain during the trial period were followed for response to treatment for 6 months. Each patient was treated with pRFA under CT-guidance with concurrent perineural injection of anaesthetic and/or corticosteroid. Pain scores were then measured using a numeric rating scale at fixed intervals up to 6 months. RESULTS: All procedures were considered technically successful with no immediate complications. pRFA demonstrated improved duration of pain improvement compared to the most recent perineural injection (p=0.0195), but not compared to the initial injection (p=0.64). Reported pain scores were lower with pRFA than with both the first and most recent injection but this did not reach statistical significance (p=0.1094 and p=0.7539, respectively). CONCLUSION: Overall, pRFA of the pudendal nerve using CT-guidance can be a safe and effective therapy. This technique provides direct visualisation of the nerve to maximise safety and efficacy while offering a novel form of therapy for patients with chronic, recalcitrant pelvic pain.

A Flow Cytometric Study of ER Stress and Autophagy.

The mechanistic link between ER stress, autophagy, and resultant cell death was investigated by the use of drugs Thapsigargin (Tg) and Chloroquine (CQ) with prior induction and or blockade of autophagy and apoptosis which modulated the ER stress response and resultant form of cell death. All these biological processes can be measured flow cytometrically allowing the determination of the type of cell death, G1 cell cycle arrest, cell cycle dependent measurement of ER stress transducer PERK, misfolded proteins, reticulophagy, and autophagy marker LC3B. Jurkat cells after Tg or CQ treatment became necrotic and apoptotic, showed G1 cell cycle arrest, autophagy, and ER stress. Prior induction of autophagy before ER stress increased levels of necrotic and apoptotic cell death. Autophagy was further up-regulated, while PERK was reduced or abrogated. CQ showed reduced levels of misfolded proteins and reticulophagy, while Tg showed no change in misfolded protein levels but increased reticulophagy and thus displayed more ER stress. Prior blockade of apoptosis before induction of ER stress resulted in cell survival except with high Tg levels which induced necrosis. Autophagy was up-regulated with modulation of PERK and reticulophagy levels with an abrogation of the misfolded protein response. Blockade of apoptosis with induction of autophagy before ER stress showed death by necrosis with high dose drugs and cell survival with low doses of drugs. CQ induced reduced levels G1 cell cycle arrest while it was maintained with Tg. Autophagy was also maintained with reduced levels of ER stress. These data demonstrates a profound link between the processes of ER stress, autophagy, and the resultant form of cell death all of which can be modulated depending upon the sequence and concentration of drugs employed. © 2018 International Society for Advancement of Cytometry.

Targeting the DNA Damage Response in OSCC with TP53 Mutations.

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer worldwide and in the United States. OSCC remains a major cause of morbidity and mortality in patients with head and neck cancers. Tobacco and alcohol consumption alone or with chewing betel nut are potential risk factors contributing to the high prevalence of OSCC. Multimodality therapies, including surgery, chemotherapy, biologic therapy, and radiotherapy, particularly intensity-modulated radiotherapy (IMRT), are the current treatments for OSCC patients. Despite recent advances in these treatment modalities, the overall survival remains poor over the past years. Recent data from whole-exome sequencing reveal that TP53 is commonly mutated in human papillomavirus-negative OSCC patients. Furthermore, these data stressed the importance of the TP53 gene in suppressing the development and progression of OSCC. Clinically, TP53 mutations are largely associated with poor survival and tumor resistance to radiotherapy and chemotherapy in OSCC patients, which makes the TP53 mutation status a potentially useful molecular marker prognostic and predictive of clinical response in these patients. Several forms of DNA damage have been shown to activate p53, including those generated by ionizing radiation and chemotherapy. The DNA damage stabilizes p53 in part via the DNA damage signaling pathway that involves sensor kinases, including ATM and ATR and effector kinases, such as Chk1/2 and Wee1, which leads to posttranscriptional regulation of a variety of genes involved in DNA repair, cell cycle control, apoptosis, and senescence. Here, we discuss the link of TP53 mutations with treatment outcome and survival in OSCC patients. We also provide evidence that small-molecule inhibitors of critical proteins that regulate DNA damage repair and replication stress during the cell cycle progression, as well as other molecules that restore wild-type p53 activity to mutant p53, can be exploited as novel therapeutic approaches for the treatment of OSCC patients bearing p53 mutant tumors.

Effect of fiber blends, total solids, heat treatment, whey protein concentrate and stage of sugar incorporation on dietary fiber-fortified Kheer.

Owing to the proven beneficial role of dietary fiber (DF) on human health, feasibility of incorporating commercially available soluble and insoluble DF preparations into rice-milk-pudding (kheer, a popular Indian delicacy) was studied through process modification. The novel approach of preparing reduced fat DF-fortified-kheer (DFFK) by developing liquid/cream phase and particulate/rice phase separately, and subsequently blending the two was developed. The major processing variables studied were total solids (TS) in the liquid phase, type of fiber blend, flavor-simulation through heat treatment or added whey protein, and the presence of sugar in water for pre-cooking of rice. Reduced fat DFFK made from three different pre-standardized fiber blends was quite acceptable to the sensory panel (overall rating 7.5). With increasing TS in milk up to 16.5%, sensory acceptability of DFFK increased. There was a small but perceivable improvement in the flavour of DFFK when precooking of rice was carried out in sweetened water. Reduced fat DFFK from different fiber blends was found to be reasonably close to conventional kheer. DFFK prepared from Blend-I provided 3.31 g dietary fiber/100 kcal (suitable for the claim "High-in-Fiber") with 38.71% Reduced-Fat. With the developed process of fortification of kheer with DF, it is possible to reduce fat (and thus calories) to cater to the needs of consumers seeking good health.

Pre-operative functional status as a predictor of morbidity and mortality after elective cervical spine surgery.

AIMS: Patients seeking cervical spine surgery are thought to be increasing in age, comorbidities and functional debilitation. The changing demographics of this population may significantly impact the outcomes of their care, specifically with regards to complications. In this study, our goals were to determine the rates of functionally dependent patients undergoing elective cervical spine procedures and to assess the effect of functional dependence on 30-day morbidity and mortality using a large, validated national cohort. PATIENTS AND METHODS: A retrospective analysis of the American College of Surgeons National Surgical Quality Improvement Program data files from 2006 to 2013 was conducted to identify patients undergoing common cervical spine procedures. Multivariate logistic regression models were generated to analyse the independent association of functional dependence with 30-day outcomes of interest. RESULTS: Patients with lower functional status had significantly higher rates of medical comorbidities. Even after accounting for these comorbidities, type of procedure and pre-operative diagnosis, analyses demonstrated that functional dependence was independently associated with significantly increased odds of sepsis (odds ratio (OR) 5.04), pulmonary (OR 4.61), renal (OR 3.33) and cardiac complications (OR 4.35) as well as mortality (OR 11.08). CONCLUSIONS: Spine surgeons should be aware of the inherent risks of these procedures with the functionally dependent patient population when deciding on whether to perform cervical spine surgery, delivering pre-operative patient counselling, and providing peri-operative management and surveillance. Cite this article: Bone Joint J 2017;99-B:824-8.

Determination of relationship between sensory viscosity rating and instrumental flow behaviour of soluble dietary fibers.

The shear-thinning low, medium and high-viscosity fiber preparations (0.15-1.05 % psyllium husk, 0.07-0.6 % guar gum, 0.15-1.20 % gum tragacanth, 0.1-0.8 % gum karaya, 0.15-1.05 % high-viscosity Carboxy Methyl Cellulose and 0.1-0.7 % xanthan gum) showed that the consistency coefficient (k) was a function of concentration, the relationship being exponential (R(2), 0.87-0.96; P < 0.01). The flow behaviour index (n) (except for gum karaya and CMC) was exponentially related to concentration (R(2), 0.61-0.98). The relationship between k and sensory viscosity rating (SVR) was essentially linear in nearly all cases. The SVR could be predicted from the consistency coefficient using the regression equations developed. Also, the relationship of k with fiber concentration would make it possible to identify the concentration of a particular gum required to have desired consistency in terms of SVR.

Clinical Characteristics and prognostic analysis of Triple-negative Breast Cancer: Single institute experience.

BACKGROUND: Triple-negative breast cancer (TNBC) is a poor prognostic subset of breast cancer that lacks the benefit of specific targeted therapy. MATERIALS AND METHODS: A prospective study of the clinical profile of triple negative breast cancer cases at a tertiary referral centre. The duration of the study period was 26 months and the median follow up period was ten months. A total of 111 invasive breast cancer patients were evaluated from 1st August 2009 to 31st October 2011. We examined TNBC patients with respect to clinicopathological parameters, adjuvant chemotherapy regimens and relapse free survival. RESULTS: In our study, patients were young (median age at presentation, 47yrs), premenopausal (54%), tumour size was discordant with lymph node positivity, the histology was predominantly intraductal carcinoma (90%), histological grade higher than two (90%). Relapses were early and preferential visceral (32%) and CNS metastasises (11.7%). 91% of patients were eligible for adjuvant therapy but only 80% of the patients could complete full course of adjuvant chemotherapy. Anthracycline-based regimens (43%), sequential anthracycline and taxane-based regimen (24%) and other regimes like CMF (13%) were used as adjuvant chemotherapy in eligible TNBC patients. Median relapse free survival in patients following adjuvant chemotherapy was around 10 months at last follow-up. CONCLUSIONS: Patients with TNBC have aggressive clinicopathological characteristics with early and higher rate of disease relapse and therefore derive inadequate benefit from current adjuvant chemotherapy. So, new treatment strategies in adjuvant chemotherapy for TNBC are needed.

Heat penetration characteristics and physico-chemical properties of in-pouch processed dairy dessert (kheer).

Kheer, a traditional milk product of South East Asia, containing cooked rice grains in a creamy sweetened concentrated milk, has no large-scale production due to its poor shelf life. Shelf-life was improved by development of a process based on in-pouch thermal processing employing a rotary retort. Product development included optimization of rice-to-milk solids ratio (0.18-0.52) and total milk solids levels (16-26 %) to simulate the conventional product in taste, appearance and textural attributes. Various Fo values (12.4-14.8) were examined with regard to product quality. While the TBA value tended to increase (0.073-0.081) the reflectance value (35.3-43.4) declined with increasing Fo. The pH of the product (6.04-6.10) showed a slight tendency to increase with Fo. Sensorily, the product was found to be acceptable for a period of 150 days at 37 °C.

Cloning of Dirac fermions in graphene superlattices.

Superlattices have attracted great interest because their use may make it possible to modify the spectra of two-dimensional electron systems and, ultimately, create materials with tailored electronic properties. In previous studies (see, for example, refs 1-8), it proved difficult to realize superlattices with short periodicities and weak disorder, and most of their observed features could be explained in terms of cyclotron orbits commensurate with the superlattice. Evidence for the formation of superlattice minibands (forming a fractal spectrum known as Hofstadter's butterfly) has been limited to the observation of new low-field oscillations and an internal structure within Landau levels. Here we report transport properties of graphene placed on a boron nitride substrate and accurately aligned along its crystallographic directions. The substrate's moiré potential acts as a superlattice and leads to profound changes in the graphene's electronic spectrum. Second-generation Dirac points appear as pronounced peaks in resistivity, accompanied by reversal of the Hall effect. The latter indicates that the effective sign of the charge carriers changes within graphene's conduction and valence bands. Strong magnetic fields lead to Zak-type cloning of the third generation of Dirac points, which are observed as numerous neutrality points in fields where a unit fraction of the flux quantum pierces the superlattice unit cell. Graphene superlattices such as this one provide a way of studying the rich physics expected in incommensurable quantum systems and illustrate the possibility of controllably modifying the electronic spectra of two-dimensional atomic crystals by varying their crystallographic alignment within van der Waals heterostuctures.

Storage related changes in ghee-based low-fat spread.

In order to assess the shelf life of a low fat spread (LFS) based on ghee, the product with (PS) and without (CS) added 0.05 % (w/w) preservative potassium sorbate and packaged in 200 g polystyrene tubs was stored at 5 °C and evaluated for changes in sensory, physico-chemical and microbiological properties. On the basis of flavour score, the PS spread could be stored for 10 weeks without appreciable loss in quality as against the CS spread which could be stored only for 5 weeks. From the point of view of spreadability, body and texture and colour, the CS product was acceptable even after 11 weeks of storage. Use of preservative had an inhibitory effect on the development of free fatty acids (FFA) and thiobarbituric acid (TBA) reactive substances. While both the products showed an increasing tendency to whey off during storage, CS wheyed off more than PS. The two spreads showed similar oiling off, which increased slightly during the storage. Microbiologically, the ghee-based low fat spreads with and without preservative was stable for 9 and 3 weeks, respectively, from the view point of yeast and mould growth; but the preservative had little effect on the total viable count. Coliforms were absent in all the samples in fresh and during storage.

Anti-infective and osteointegration properties of silicon nitride, poly(ether ether ketone), and titanium implants.

Silicon nitride (Si(3)N(4)) is an industrial ceramic used in spinal fusion and maxillofacial reconstruction. Maximizing bone formation and minimizing bacterial infection are desirable attributes in orthopedic implants designed to adhere to living bone. This study has compared these attributes of Si(3)N(4) implants with implants made from two other orthopedic biomaterials, i.e. poly(ether ether ketone) (PEEK) and titanium (Ti). Dense implants made of Si(3)N(4), PEEK, or Ti were surgically implanted into matching rat calvarial defects. Bacterial infection was induced with an injection of 1×10(4)Staphylococcus epidermidis. Control animals received saline only. On 3, 7, and 14days, and 3months post-surgery four rats per time period and material were killed, and calvariae were examined to quantify new bone formation and the presence or absence of bacteria. Quantitative evaluation of osteointegration to adjacent bone was done by measuring the resistance to implant push-out (n=8 rats each for Ti and PEEK, and n=16 rats for Si(3)N(4)). Three months after surgery in the absence of bacterial injection new bone formation around Si(3)N(4) was ∼69%, compared with 24% and 36% for PEEK and Ti, respectively. In the presence of bacteria new bone formation for Si(3)N(4), Ti, and PEEK was 41%, 26%, and 21%, respectively. Live bacteria were identified around PEEK (88%) and Ti (21%) implants, whereas none were present adjacent to Si(3)N(4). Push-out strength testing demonstrated statistically superior bone growth onto Si(3)N(4) compared with Ti and PEEK. Si(3)N(4) bioceramic implants demonstrated superior new bone formation and resistance to bacterial infection compared with Ti and PEEK.

Diffusion-weighted imaging in characterization of cystic pancreatic lesions.

AIM: To evaluate whether apparent diffusion coefficient (ADC) measurements from diffusion-weighted imaging (DWI) can characterize or predict the malignant potential of cystic pancreatic lesions. MATERIALS AND METHODS: Retrospective review of the magnetic resonance imaging (MRI) database over a 2-year period revealed 136 patients with cystic pancreatic lesions. Patients with DWI studies and histological confirmation of cystic mass were included. In patients with known pancreatitis, lesions with amylase content of >1000 IU/l that resolved on subsequent scans were included as pseudocysts. ADC of cystic lesions was measured by two independent reviewers. These values were then compared to categorize these lesions as benign or malignant using conventional MRI sequences. RESULTS: Seventy lesions were analysed: adenocarcinoma (n=4), intraductal papillary mucinous neoplasm (IPMN; n=28), mucinous cystic neoplasm (MCN; n=9), serous cystadenoma (n=16), and pseudocysts (n=13). There was no difference between ADC values of malignant and non-malignant lesions (p=0.06), between mucinous and serous tumours (p=0.12), or between IPMN and MCN (p=0.42). ADC values for low-grade IPMN were significantly higher than those for high-grade or invasive IPMN (p=0.03). CONCLUSION: ADC values may be helpful in deciding the malignant potential of IPMN. However, they are not useful in differentiating malignant from benign lesions or for characterizing cystic pancreatic lesions.

A comparative study of kinetic and connectionist modeling for shelf-life prediction of Basundi mix.

A ready-to-reconstitute formulation of Basundi, a popular Indian dairy dessert was subjected to storage at various temperatures (10, 25 and 40 °C) and deteriorative changes in the Basundi mix were monitored using quality indices like pH, hydroxyl methyl furfural (HMF), bulk density (BD) and insolubility index (II). The multiple regression equations and the Arrhenius functions that describe the parameters' dependence on temperature for the four physico-chemical parameters were integrated to develop mathematical models for predicting sensory quality of Basundi mix. Connectionist model using multilayer feed forward neural network with back propagation algorithm was also developed for predicting the storage life of the product employing artificial neural network (ANN) tool box of MATLAB software. The quality indices served as the input parameters whereas the output parameters were the sensorily evaluated flavour and total sensory score. A total of 140 observations were used and the prediction performance was judged on the basis of per cent root mean square error. The results obtained from the two approaches were compared. Relatively lower magnitudes of percent root mean square error for both the sensory parameters indicated that the connectionist models were better fitted than kinetic models for predicting storage life.

Evaluation of the subaxial injury classification system.

STUDY DESIGN: Retrospective clinical study of patients treated for subaxial cervical spine trauma (SCST) at a tertiary medical center. PURPOSE: Evaluate the validity of the Subaxial Injury Classification (SLIC) system in surgical versus non-surgical decision making for SCST. INCLUSION CRITERIA: Age >12 years, presence of SCST with complete clinical and radiological (CT and MRI) data. EXCLUSION CRITERIA: Patients with incomplete radiographic or clinical data, pathological fractures, isolated upper cervical trauma (occiput to C2), isolated transverse process or spinous process fractures, chronic or age indeterminate fractures, isolate MRI findings, and severe systemic trauma with death prior to either surgical or non-surgical treatment. RESULTS: Fourteen patients were treated non-surgically (C), whereas 24 were treated surgically (S). In the C group, the SLIC score ranged from 0 to 5 points (standard deviation [SD] = 1.20 points; mean 1.07; median 1). Just 1 patient had an SLIC score greater than 2 (7.1% of the patients). In the S group, the SLIC score ranged from 1 to 10 points (standard deviation [SD] = 2.03 points; mean 5.6; median 6). Just 2 patients had an SLIC score smaller than 4 (both with 1 point each, 8.3% of the total group). All the other 22 (accounting for 91.6%) patients had an SLIC of 4 or more points. CONCLUSIONS: Our study suggests that the SLIC classification looks to be a promising system to aid spinal surgeons in the decision-making process of subaxial cervical trauma, but a large prospective cohort study is required.

When is it reasonable to split pediatric en bloc kidneys for transplantation into two adults?

BACKGROUND: Traditionally, kidneys from donors ≥60 years old and pediatric kidneys are considered marginal organs for transplantation. Pediatric donor kidneys are underutilized for transplantation into adult recipients due to concern for poor outcomes. METHODS: Using data from the Organ Procurement and Transplant Network, we analyzed patterns of pediatric kidney use (single vs en bloc) in the United States from 1987 to 2007. Using the Cox proportional hazards model, graft outcomes of pediatric donor kidneys transplanted as single vs en bloc grafts from different donor weight groups were compared with renal transplantation from donors ≥60 years old in an attempt to define a pediatric donor weight at which kidneys can be justifiably split to expand the donor pool. RESULTS: Compared with older donor kidneys, graft failure risk of pediatric single kidneys was consistently lower when the donor weight exceeded 10 kg. On the other hand, graft survival benefit for pediatric en bloc kidneys was evident starting at donor weight ≤10 kg in comparison to older donor kidneys. Pediatric en bloc kidneys performed consistently better than pediatric single kidneys for all donor weight groups. CONCLUSIONS: Splitting of pediatric donor en bloc kidneys for transplantation into 2 adults when the donor weight exceeds 10 kg was associated with acceptable graft outcomes. This practice, along with increased use of small pediatric donor kidneys, may help to alleviate the waiting list burden in renal transplantation.

Quantification of diffusivities of the human cervical spinal cord using a 2D single-shot interleaved multisection inner volume diffusion-weighted echo-planar imaging technique.

BACKGROUND AND PURPOSE: DTI is a highly sensitive technique, which can detect pathology not otherwise noted with conventional imaging methods. This paper provides the atlas of reliable normative in vivo DTI parameters in the cervical spinal cord and its potential applications toward quantifying pathology. MATERIALS AND METHODS: In our study, we created a reference of normal diffusivities of the cervical spinal cord by using a 2D ss-IMIV-DWEPI technique from 14 healthy volunteers and compared parameters with those in 8 patients with CSM. The 2D ss-IMIV-DWEPI technique was applied in each subject to acquire diffusion-weighted images. FA, lambda( parallel), and lambda( perpendicular) were calculated. A reference of normal DTI indices from 12 regions of interest was created and compared with DTI indices of 8 patients. RESULTS: A map of reference diffusivity values was obtained from healthy controls. We found statistically significant differences in diffusivities between healthy volunteers and patients with CSM with different severities of disease, by using FA, lambda( parallel), and lambda( perpendicular) values. CONCLUSIONS: DTI using 2D ss-IMIV-DWPEI is sensitive to spinal cord pathology. This technique can be used to detect and quantify the degree of pathology within the cervical spinal cord from multiple disease states.

Intensive glucose control and macrovascular outcomes in type 2 diabetes.

AIMS/HYPOTHESIS: Improved glucose control in type 2 diabetes is known to reduce the risk of microvascular events. There is, however, continuing uncertainty about its impact on macrovascular disease. The aim of these analyses was to generate more precise estimates of the effects of more-intensive, compared with less-intensive, glucose control on the risk of major cardiovascular events amongst patients with type 2 diabetes. METHODS: A prospectively planned group-level meta-analysis in which characteristics of trials to be included, outcomes of interest, analyses and subgroup definitions were all pre-specified. RESULTS: A total of 27,049 participants and 2,370 major vascular events contributed to the meta-analyses. Allocation to more-intensive, compared with less-intensive, glucose control reduced the risk of major cardiovascular events by 9% (HR 0.91, 95% CI 0.84-0.99), primarily because of a 15% reduced risk of myocardial infarction (HR 0.85, 95% CI 0.76-0.94). Mortality was not decreased, with non-significant HRs of 1.04 for all-cause mortality (95% CI 0.90-1.20) and 1.10 for cardiovascular death (95% CI 0.84-1.42). Intensively treated participants had significantly more major hypoglycaemic events (HR 2.48, 95% CI 1.91-3.21). Exploratory subgroup analyses suggested the possibility of a differential effect for major cardiovascular events in participants with and without macrovascular disease (HR 1.00, 95% CI 0.89-1.13, vs HR 0.84, 95% CI 0.74-0.94, respectively; interaction p = 0.04). CONCLUSIONS/INTERPRETATION: Targeting more-intensive glucose lowering modestly reduced major macrovascular events and increased major hypoglycaemia over 4.4 years in persons with type 2 diabetes. The analyses suggest that glucose-lowering regimens should be tailored to the individual.