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The increasing demand for cryo-electron microscopy (cryo-EM) reveals drawbacks in current sample preparation protocols, such as sample waste and lack of reproducibility. Here, we present several technical developments that provide efficient sample preparation for cryo-EM studies. Pin printing substantially reduces sample waste by depositing only a sub-nanoliter volume of sample on the carrier surface. Sample evaporation is mitigated by dewpoint control feedback loops. The deposited sample is vitrified by jets of cryogen followed by submersion into a cryogen bath. Because the cryogen jets cool the sample from the center, premounted autogrids can be used and loaded directly into automated cryo-EMs. We integrated these steps into a single device, named VitroJet. The device’s performance was validated by resolving four standard proteins (apoferritin, GroEL, worm hemoglobin, beta-galactosidase) to ~3 Å resolution using a 200-kV electron microscope. The VitroJet offers a promising solution for improved automated sample preparation in cryo-EM studies. There is a need to further improve the automation of cryo-EM sample preparation to make it more easily accessible for non-specialists, reduce sample waste and increase reproducibility. Here, the authors present VitroJet, a single device, where sub-nl volumes of samples are deposited by pin printing thus eliminating the need for sample blotting, which is followed by jet vitrification, and they show that high-resolution structures can be obtained using four standard proteins.

There is a well recognized association between depression and diabetes. However, there is little empirical data about the prevalence of depressive symptoms and anxiety among different groups of glucose metabolism in population based samples. The aim of this study was to determine whether the prevalence of increased levels of depression and anxiety is different between patients with type 2 diabetes and subjects with impaired glucose metabolism (IGM) and normal glucose metabolism (NGM). Cross-sectional data from a population-based cohort study of 2667 residents, 1261 men and 1406 women aged 40-65 years from the Hoorn region, the Netherlands. Depressive symptoms and anxiety were measured using the Centre for Epidemiologic Studies Depression Scale (CES-D, score >or=16) and the Hospital Anxiety and Depression Scale--Anxiety Subscale (HADS-A, score >or=8), respectively. Glucose metabolism status was determined by oral glucose tolerance test. In the total study population the prevalence of depressive symptoms and anxiety for the NGM, IGM and type 2 diabetes were 12.5, 12.2 and 21.0% (P = 0.004) and 15.0, 15.3 and 19.9% (p = 0.216), respectively. In men, the prevalence of depressive symptoms was 7.7, 9.5 and 19.6% (p<0.001), and in women 16.4, 15.8 and 22.6 (p = 0.318), for participants with NGM, IGM and type 2 diabetes, respectively. Anxiety was not associated with glucose metabolism when stratified for sex. Intergroup differences (NGM vs. IGM and IGM vs. type 2 diabetes) revealed that higher prevalences of depressive symptoms are mainly manifested in participants with type 2 diabetes, and not in participants with IGM. Depressive symptoms, but not anxiety are associated with glucose metabolism. This association is mainly determined by a higher prevalence of depressive symptoms in participants with type 2 diabetes and not in participants with IGM.

Diabetes-Related Symptom Distress in Association With Glucose Metabolism and Comorbidity The Hoorn Study Marcel C. Adriaanse , PHD 1 , Frans Pouwer , PHD 2 3 , Jacqueline M. Dekker , PHD 2 , Giel Nijpels , MD, PHD 2 4 , Coen D. Stehouwer , MD, PHD 5 , Robert J. Heine , MD, PHD 2 6 and Frank J. Snoek , PHD 2 3 1 Institute of Health Sciences, Faculty of Earth and Life Sciences, VU University Amsterdam, Amsterdam, the Netherlands 2 EMGO Institute, VU University Medical Centre, Amsterdam, the Netherlands 3 Department of Medical Psychology, VU University Medical Centre, Amsterdam, the Netherlands 4 Department of General Practice, VU University Medical Centre, Amsterdam, the Netherlands 5 Department of Internal Medicine, Academic Hospital Maastricht, Maastricht, the Netherlands 6 Department of Endocrinology, VU University Medical Centre, Amsterdam, the Netherlands Corresponding author: Marcel C. Adriaanse, marcel.adriaanse{at}falw.vu.nl Abstract OBJECTIVE —The purpose of this study was to determine the associations between diabetes-related symptom distress, glucose metabolism status, and comorbidities of type 2 diabetes. RESEARCH DESIGN AND METHODS —This was a cross-sectional sample of 281 individuals with normal glucose metabolism (NGM), 181 individuals with impaired glucose metabolism (IGM), and 107 subjects with type 2 diabetes. We used the revised type 2 Diabetes Symptom Checklist (DSC-R) to assess diabetes-related symptom distress. RESULTS —The total symptom distress score (range 0–100) was relatively low for diabetic subjects (mean ± SD 8.4 ± 9.4), although it was significantly different from that for subjects with IGM (6.5 ± 7.1) and NGM (6.1 ± 7.9) ( F = 3.1, 2 d.f., P = 0.046). Ischemic heart disease was associated with elevated DSC-R scores on three subscales, whereas depression showed higher symptom distress levels across all DSC-R domains. CONCLUSIONS —Worsening glucose metabolism is associated with increasing diabetes-related symptom distress. This relationship is attenuated by ischemic heart disease and particularly by depression. Footnotes Published ahead of print at http://care.diabetesjournals.org on 26 August 2008. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Accepted August 15, 2008. Received June 13, 2008. DIABETES CARE

The identification and discussion of sexual care needs in people with type 2 diabetes mellitus (T2DM) in primary care is currently insufficient. The objective of this study was to determine the prevalence of sexual dissatisfaction, sexual problems and need for help by using a screening instrument among people with T2DM in primary care. Data were collected in 45 general practices in the Netherlands from January 2015 to February 2016. The Brief Sexual Symptom Checklist (BSSC) was used to screen among 40–75 year old men and women. In total, 786 people with T2DM (66.5% men) were screened. The prevalence of sexual dissatisfaction was 36.6%, significantly higher among men than among women (41.1% vs. 27.8%). Sexually dissatisfied men most often reported erectile dysfunction (71.6%); for sexually dissatisfied women, low sexual desire (52.8%) and lubrication problems (45.8%) were most common. More than half of all dissatisfied people had a need for care (61.8%), significantly more men than women (66.8% vs. 47.2%). One third of people with T2DM is sexually dissatisfied and more than half of these people report a need for help. The BSSC could be used a tool to proactively identify sexually dissatisfied people in primary care. •Sexual problems are one of the most frequently neglected complications in diabetes care.•The use of a screening instrument could identify people with sexual problems in need for help.•The Brief Sexual Symptom Checklist (BSSC) was used to screen among 786 men and women with type 2 diabetes in primary care.•• One third of people were sexually dissatisfied and more than half of these people reported a need for help.•The BSSC could be used a tool to proactively identify sexually dissatisfied people in primary care.

OBJECTIVE:--The purpose of this study was to determine the associations between diabetes-related symptom distress, glucose metabolism status, and comorbidities of type 2 diabetes. RESEARCH DESIGN AND METHODS--This was a cross-sectional sample of 281 individuals with normal glucose metabolism (NGM), 181 individuals with impaired glucose metabolism (IGM), and 107 subjects with type 2 diabetes. We used the revised type 2 Diabetes Symptom Checklist (DSC-R) to assess diabetes-related symptom distress. RESULTS:--The total symptom distress score (range 0-100) was relatively low for diabetic subjects (mean ± SD 8.4 ± 9.4), although it was significantly different from that for subjects with IGM (6.5 ± 7.1) and NGM (6.1 ± 7.9) (F = 3.1, 2 d.f., P = 0.046). Ischemic heart disease was associated with elevated DSC-R scores on three subscales, whereas depression showed higher symptom distress levels across all DSC-R domains. CONCLUSIONS:--Worsening glucose metabolism is associated with increasing diabetes-related symptom distress. This relationship is attenuated by ischemic heart disease and particularly by depression.

Objective: To determine the level of diabetes-related symptom distress and its association with negative mood in subjects participating in a targeted population-screening program, comparing those identified as having type 2 diabetes vs. those who did not. Research design and methods: This study was conducted within the framework of a targeted screening project for type 2 diabetes in a general Dutch population (age 50-75 years). The study sample consisted of 246 subjects, pre-selected on the basis of a high-risk profile; 116 of whom were subsequently identified as having type 2 diabetes, and 130 who were non-diabetic subjects. Diabetes-related symptom distress and negative mood was assessed ∼2 weeks, 6 months, and 12 months after the diagnosis of type 2 diabetes, with the Type 2 Diabetes Symptom Checklist and the Negative well-being sub scale of the Well-being Questionnaire (W-BQ12), respectively. Results: Screening-detected diabetic patients reported significantly greater burden of hyperglycemic (F = 6.0, df = 1, p = 0.015) and of fatigue (F = 5.3, df = 1, p = 0.023) symptoms in the first year following diagnosis type 2 diabetes compared to non-diabetic subjects. These outcomes did not change over time. The total symptom distress (range 0-4) was relatively low for both screening-detected diabetic patients (median at ∼2 weeks, 6 months, and 12 months; 0.24, 0.24, 0.29) and non-diabetic subjects (0.15, 0.15, 0.18), and not significantly different. No average difference and change over time in negative well-being was found between screening-detected diabetic patients and non-diabetic subjects. Negative well-being was significantly positive related with the total symptom distress score (regression coefficient β = 2.86, 95% CI 2.15-3.58). Conclusions: The screening-detected diabetic patients were bothered more by symptoms of hyperglycemia and fatigue in the first year following diagnosis type 2 diabetes than non-diabetic subjects. More symptom distress is associated with increased negative mood in both screening-detected diabetic patients and non-diabetic subjects.

The increasing demand for cryo-electron microscopy (cryo-EM) reveals drawbacks in current sample preparation protocols, such as sample waste and lack of reproducibility. Here, we present several technical developments that provide controlled and efficient sample preparation for cryo-EM studies. Pin printing substantially reduces sample waste by depositing only a sub-nanoliter volume of sample on the carrier surface. Sample evaporation is mitigated by dewpoint control feedback loops. The deposited sample is vitrified by jets of cryogen followed by submersion into a cryogen bath. Because the cryogen jets cool the sample from the center, premounted autogrids can be used and loaded directly into automated cryo-EMs. We integrated these steps into a single device, named VitroJet. The device's performance was validated by resolving 4 standard proteins (apoferritin, GroEL, worm hemoglobin, beta-galactosidase) to ~3 Å resolution using a 200-kV electron microscope. The VitroJet offers a promising solution for improved automated sample preparation in cryo-EM studies.