We investigated the vision-related quality of life (VR-QOL) in patients with HLA-B27 associated anterior uveitis (AU). The study was conducted in 2012 at the ophthalmology department of the University Medical Center of Groningen. We included AU patients who were HLA-B27 positive and/or were diagnosed by a rheumatologist with an HLA-B27 associated systemic disease. Sixty-one of 123 (50%) adult patients participated. All patients filled-out the National Eye Institute Visual Functioning Questionnaire-25 (NEI VFQ-25), Beck Depression Inventory (BDI-II), social support lists and an additional questionnaire for gathering general information. Medical records were reviewed for clinical characteristics. Analyses were conducted on various patient and ocular characteristics. We compared our NEI VFQ-25 scores with those previously found in the literature. Our main outcome measures were VR-QOL scores and their associations with various general patient and ocular characteristics. We found that the NEI VFQ-25 mean overall composite score was 88.9±8.8, which is relatively high, but lower than that found in a normal working population. The mean general health score was 47.4±20.8, which is lower than in patients with other ocular diseases. Patients with a systemic disease scored significantly lower on general health and VR-QOL, compared to patients without a systemic disease. Patients with a depression (6/59 (10%)) frequently had ankylosing spondylitis (5/6 patients) and they scored significantly worse on VR-QOL. We concluded that patients with HLA-B27 associated AU have a relatively high VR-QOL. However, the presence of a systemic disease is associated with lower VR-QOL and general health scores. In addition, depression is associated with a lower VR-QOL. Anterior uveitis (AU) is the most common form of uveitis and it is commonly associated with HLA-B27 associated diseases, such as ankylosing spondilitis, Crohn’s disease, reactive arthritis and psoriasis. Also, HLA-B27 positivity without apparent systemic disease may be associated with AU. This typPrevious studies indicated a poorer visual functioning and a lower general health status in uveitis patients compared to healthy subjects. Most stuThe purpose of the present study is to evaluate the VR-QOL in a group of patients with an HLA-B27 associated AU. We hope to identify clinical features that are associated with a lower quality of life in this patient group. Identifying such features may help to develop targeted screening and intervention measures in the future.Medical Research Involving Human Subjects Act. The study was conducted according to the tenets of the Declaration of Helsinki.The Medical Ethical Committee of the University Medical Center of Groningen ruled that approval was not required for this study, since the study is not a medical scientific study with people as defined in the We selected all patients from an existing database, containing uveitis patients who are currently being treated or who have been treated for uveitis at the ophthalmology department of the University Medical Center Groningen, which is a tertiary referral center.We identified 123 adult (18 years or older) patients with HLA-B27 associated AU. Patients were HLA-B27 positive and/or were clinically diagnosed with an HLA-B27 associated systemic disease by a rheumatologist. Patients with ankylosing spondylitis were diagnosed following the Assessment of SpondyloArthritis international Society classification criteria (ASAS criteria). Therefore patients with sacroiliitis on imaging and ≥ one axial spondyloarthritis criteria, have not always been tested for HLA-B27 positivity. [All suitable 123 patients received the following questionnaires by mail: the National Eye Institute Visual Functioning Questionnaire-25 (NEI VFQ-25), the Beck Depression Inventory (BDI-II), Social Support List–Interactions (SSL-I), Social Support List–Discrepancies (SSL-D) and an additional questionnaire for gathering general information. They also received an information letter and an informed consent form. Patients could complete the questionnaires and sign the informed consent form at home and return them by mail. We contacted all patients by phone, to ask if they needed assistance in filling out the questionnaire. All patients were literate.In this study, we used the Dutch version of the NEI VFQ-25. This validated ,10 questThe BDI-II is a validated  self-admThe SSL-I and SSL-D are questionnaires developed and validated by the University of Groningen. They each consist of 34 four-choice questions, resulting in scores ranging from 1–4. The SSL-I questionnaire measures the social interactions between patients and persons with whom they interact. The maximum score is 136, a high SSL-I score corresponds with sufficient social support. The SSL-D questionnaire measures if the received social support corresponds with the desired social support. The maximum score is 102, a high SSL-D score corresponds with a deficiency in desired social support.The additional questionnaire gave us the following information: whether the uveitis was active at the time of completing the questionnaire, medical history , medication use (ocular and other medication) and history of depression and the need for treatment.The following information was collected by examining medical records: current age, sex, uni- or bilateral AU, presence of a systemic disease, follow-up (time between the start of the first uveitis episode and the end of the last uveitis episode), total number of uveitis episodes, total time of active disease in months, remission time (time between the end of the last uveitis episode and the date on the questionnaire), Snellen VA, ocular complications (elevated intraocular pressure (IOP), glaucoma, cataract, secondary cataract, dry eyes and current treatment for dry eyes, cystoid macular edema (CME), papillitis, scleritis and posterior synechiae) and present activity of the uveitis. To evaluate a possible bias by inclusion, the following data were collected from the patients who did not return the questionnaire: visual acuity at the end of follow-up, duration of follow-up, presence of systemic disease, bilaterality of disease  and ocular complications.Active uveitis was defined as ≥0.5+ cells in the anterior chamber. TransienData were statistically analyzed using SPSS Statistics 20.0.0.1. The Mann-Whitney U test was used to compare continuous variables of two groups. The Kruskal–Wallis one-way analysis was used to compare continuous variables of more than two groups and the Mann-Whitney U test for post hoc analysis with a Bonferroni correction, using a critical value of 0.05 divided by the number of tests conducted. Correlations were assessed with the Spearman’s Rank Correlations test. For analyzing, Snellen VA was converted to the logarithm of the minimum angle of resolution (logMAR) equivalent. Statistical significance level was set at 0.05.Sixty-one of 123 (50%) patients filled out the questionnaires and returned them by mail.The mean (± SD) of the OCS and the subscales of the NEI VFQ-25 in relation to diverse patients’ characteristics and ocular variables are shown in Tables LogMAR VA higher than 0.15 , treatment of the uveitis at the time of completing the NEI VFQ-25, presence of a systemic disease and presence of a depression at the time of completing the NEI VFQ-25 were patients’ characteristics that were highly associated with a lower NEI VFQ-25 OCS and/or NEI VFQ-25 subscales. LogMAR VA higher than 0.15 in at least one eye, was correlated to significantly lower scores on general vision, distance activities, vision specific role difficulties, vision specific dependency and color vision. Treatment of the AU and treatment of ophthalmic complications at the moment of completing the NEI VFQ-25 were related to lower scores on the subscales, vision specific social functioning, vision specific mental health, vision specific role difficulties, vision specific dependency, color vision and peripheral vision. Patients with a systemic disease scored significantly lower on the OCS and on the subscales general health, ocular pain, distance activities, vision specific mental health and peripheral vision. Depression at the time of completing the NEI VFQ-25 was correlated to significantly worse scores on the OCS and on the subscales general vision, near activities, distance activities, vision specific social functioning and driving. These patients also scored lower on all other subscales, except color vision, but these subscales did not reach significance.The patients’ characteristics that were moderately associated with lower NEI VFQ-25 OCS and/or NEI VFQ-25 subscales were older age, female gender, multiple uveitis episodes, bilateral AU, dry eyes and presence of CME. Younger patients (<45 years) scored significantly better (p = 0.026) on near activities compared to older patients (≥45 years). Post hoc analyses showed that near activity scores in patients <45 years versus those aged 45–65 years and patients <45 years versus those aged >65 years significantly differ , whereas near activities scores did not significantly differ for patients between 45–65 years and patients >65 years (p = 0.714). Female patients scored significantly worse on driving. Patients who experienced only one uveitis episode had significantly higher general vision scores, compared with patients who experienced multiple uveitis episodes. Patients with bilateral AU scored significantly lower on ocular pain (indicating that they experienced more pain or discomfort around or in the eye). Patients with dry eyes scored significantly lower on ocular pain and on the OCS. Patients with CME scored lower on vision specific dependency and color vision. Elevated IOP, posterior synechiae and treatment of the systemic disease at the moment of completing the questionnaire showed no significant correlation with any of the NEI VFQ-25 outcomes.The results of the Spearman’s Rank Correlations tests between NEI VFQ-25 subscale scores and OCS and of various patient and ocular characteristics are given in Higher LogMAR VA of the worst eye was negatively correlated with general vision and vision specific dependency. Follow-up time was negatively correlated with general vision and vision specific dependency. The BDI-II score was negatively correlated with the OCS and almost all subscales, except general vision and color vision. The SSL-D score was negatively correlated with the OCS and near activities, distance activities and vision specific role difficulties. Positive and negative correlations were seen with higher LogMAR VA of the best eye. Higher LogMAR VA of the best eye was negatively correlated with distance activities, vision specific dependency, peripheral vision and positively correlated with ocular pain. There was no significant correlation between the subscale scores or OCS and the number of uveitis episodes, duration of active uveitis and SSL-I score. The BDI-II score was positively correlated with the SSL-D score .In general, the NEI VFQ-25 OCS and subscale scores were relatively high in patients with HLA-B27 associated AU. The mean OCS was 88.9 in the total group. The best score to achieve on each question was 100 and the second best score 75 or 80. This means that most patients scored between the best and second best scores. The OCS is the average of all vision-targeted subscale scores, only excluding general health. The mean general health score was rather low, in comparison to the vision-targeted subscales, namely 47.4. This means that the majority of patients scored their general health between 'fair' (25.0) and 'good' (50.0). Six (10%) of patients had a mild depression.Patients who were currently being treated for uveitis or ophthalmic complications had lower scores on vision specific social functioning, vision specific mental health, vision specific role difficulties and vision specific dependency. This means that these patients are more worried and frustrated by their eyesight and they need more help because of their eyesight. In our patients, a history of dry eyes seemed to have no effect on the VR-QOL, patients with dry eyes in their history even scored higher on ocular pain (indicating that they experienced less pain or discomfort around or in the eye). In seeming contrast, Li et al. reported that VR-QOL in dry eye patients was significantly impaired. The difference is probably due to the fact that in Li’s study, all patients had dry eyes at the moment of completing the NEI VFQ-25. Whereas Subgroup analyses further show that younger patients (<45 years) score better on near activities compared to older patients  in our patient group was relatively low. By comparison, mean general health scores in a normal working population with and without ocular disease were 68.6 and 79.9, respectively. Patients7.4 in ouIn our patient group, patients had a mean BDI-ll score of 4.7  and six (10%) patients had a mild depression at the time of completing the questionnaire. The majority of these, (5/6 (83%)) had ankylosing spondylitis, suggesting that the presence of a systemic disease has a higher influence on this outcome than the presence of AU. Previous studies on HLA-B27 associated diseases, give similar prevalence numbers, since they describe a clinically relevant depression in 12.3% of 171 HLA-B27 associated AU patients and in 14.8% of 55 ankylosing spondylitis patients, respectively.,22 DepreThe main shortcoming of our study is its modest sample size. Our sample size is considered adequate for overall analyses, but it In conclusion, patients with HLA-B27 associated AU have a relatively high VR-QOL. However, the presence of a systemic disease is associated with considerably lower VR-QOL scores and general health scores and may be associated with an increased risk of depression. In addition, depression itself is associated with a lower VR-QOL.


Apoptosis plays a critical role in controlling the proliferation and differentiation of germ cells during spermatogenesis. Dysregulation of the fine-tuned balance may lead to the onset of testicular diseases. In this study, we investigated the activation status of apoptosis pathways in the testicular tissues under the background of an asthmatic mouse model.Ten BALB/c mice were divided into two groups: the acute asthma group and the control group. In the acute asthma group, ovalbumin (OVA)-sensitized mice were challenged with aerosolized OVA for 7 days, while the control group was treated with physiological saline. After that, both epididymis and testis were collected to determine the sperm count and motility. Apoptosis in the testis was evaluated by DNA ladder, immunochemistry and further by PCR array of apoptosis-related genes. Finally, the cleavage of caspase-3 and poly ADP-ribose polymerase (PARP) was determined by western blot and the enzymatic activities of caspase-9 and 3/7 were assessed using Caspase-Glo kits.Compared with control mice, significant decreases in the body weight, testis weight, sperm count and motility were seen in the experimental group. DNA ladder and immunochemistry showed significant increase in apoptotic index of the asthmatic testis, whereas a decrease in mRNA expression of Bcl-2 and increases in Bax, BNIP3, caspase-9, and AIF were observed in the asthma group. Furthermore, protein levels of AIF were significantly upregulated, while the translational expression of Bcl-2 was downregulated markedly. Consistently, caspase-9 activity in the testis of asthma mice was significantly higher than that of the control group.Collectively, these results showed that Bcl-2-caspase-9 apoptosis pathway was clearly activated in the testis of asthmatic mice with the increased expression of apoptosis-related genes and proteins. To our knowledge, this is the first report demonstrating that asthma could lead to the activation of the mitochondrial apoptosis signaling pathway in the mouse testis. Asthma is a common respiratory disorder with significant morbidity and mortality worldwide. It is estimated that as many as 300 million people of all ages and all ethnic backgrounds suffer from asthma ,2. CharaIn the past several decades, apoptosis has been shown to play many critical roles in diverse physiological and pathological processes. Classically, it can be activated through two distinct pathways, the death receptor pathway (extrinsic) and the mitochondrial (intrinsic) pathway. During the processes of embryonic development and cell differentiation, the mitochondrial apoptosis pathway can be activated by multiple mechanisms. Similarly, it was demonstrated that the intrinsic apoptosis pathway is also activated during spermatogenesis in the testis . It has A recent study demonstrates that time to pregnancy is prolonged in asthmatic females, especially in women with moderate to severe asthma due to unknown reasons . HoweverMice were handled in accordance with the guidelines of the Ethics Committee of Weifang People’s Hospital  and all experiments were carried out in conformity to the Guiding Principles for the Care and Use of Laboratory Animals . BALB/c mice of male, 6 to 8 weeks old , were housed in specific pathogen-free conditions at Weifang Medical University. The mice were kept in an air-conditioned room with temperatures between 20–25°C and 50–70% humidity and acclimated for 2 weeks before experiment.In order to generate the acute asthma model, acclimated mice were treated according to previously reported methods . Briefly2 asphyxiation. Immediately after being removed and weighed, the testes were frozen and stored in liquid nitrogen for real-time qPCR and western blotting.Each mouse was weighed before execution by COAs previously described , in ordeThe mice testes were removed and fixed in 4% paraformaldehyde. Paraffin-embedded sections were made and stained with hematoxylin and eosin (HE). For apoptosis analysis, the transverse sections were incubated with cleaved-caspase-3 antibody  at 4°C overnight. The apoptotic cells were stained to brown.For RNA purification, frozen testis was homogenized by liquid nitrogen grinding and dissolved in Buffer RTK provided in the Qiagen RNeasy Mini Kit. RNA was extracted according to the manufacturer's instructions. All RNA samples were suspended in RNase-free water provided with the RNA isolation kit. RNA samples were treated with RNase-free DNase I (Ambion) to digest any residual chromosomal DNA. The concentration and purity of the RNA was checked with an ultraviolet (UV) spectrophotometer at wavelengths of 260 nm and 280 nm.For PCR array, equal amounts of RNA from 5 control or experiment mice were pooled and mixed well before cDNA synthesis. Reverse transcription was carried out using PrimeScript RT Master Mix (Takara) following the manufacturer's instructions. Real-time qPCR for PCR array analysis was performed in the StepOnePlus Real-Time qPCR System (Applied Biosystems) with the SYBR Green Real-Time PCR Master Mix (Target Bio) according to the manufacturer's instructions. Duplicate plates were tested for each condition and were compared to assess reproducibility of the results. The threshold cycle (Ct) for each well was calculated using the instrument's software, and the melting-curve program was immediately run after the cycling program. The amplification efficiency of all primers was tested to be between 1.90 and 2.10. Data analysis was run by the ΔΔCt method.For confirmation of differentially expressed genes by qPCR, equal amounts of RNA samples from the 10 mouse testes were aliquoted individually and then subjected to cDNA synthesis respectively. The mRNA expression levels of HIF-1α, Bax, Bcl-2, BNIP3, AIF, cytochrome c, caspase-9 and caspase-3 were analyzed by real-time quantitative PCR using the StepOnePlus Real-Time qPCR System (Applied Biosystems). All values were normalized to the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA. The primers used were listed in Briefly, frozen testis were homogenized in RIPA buffer supplemented with protease inhibitors using the liquid nitrogen grinding, followed by incubation on ice for 10 min. The samples were centrifuged thoroughly to obtain protein supernatants. The protein concentrations were determined using a BCA Protein Assay Kit (Pierce). Twenty mg protein for each sample were resolved on 12% Bis-Tris or 4–12% gradient Bis/Tris gels (Life Technologies) and then transferred to PVDF membranes (Millipore). After blocking in 10% skim milk, the immunoblotting membrane was probed with indicated antibodies and visualized by ECL kit (Pierce). Lastly, images of indicated protein bands were recorded on the BioMax film (Kodak), and quantification was conducted by using Image J software (Bio-Rad). Primary antibodies used in the study were diluted as follows: anti-Bcl-2 , anti-PARP , anti-Bax , anti-cytochrome c , anti-HIF-1α , anti-AIF , anti-GAPDH , and anti-cleaved caspase-3 .DNA ladder assay for apoptosis detection was conducted following a recently updated protocol . After DThe activity of caspase-9 and caspase-3/7 was measured by Caspase-Glo-9 assay kit and Caspase-Glo-3/7 assay kit, respectively, according to the manufacturer's instructions (Promega). Briefly, protein concentration of the testis homogenates was determined for each mouse and equal amounts of the homogenates were subjected to the enzymatic assay following the standard protocol.All quantified data were expressed as means ± standard deviation (SD). Statistical analysis was performed using the SPSS statistical software package, version 17.0 . Significance was set at the level of P < 0.05.Mice rendered allergic to OVA have been widely studied, and they serve as a prototypic asthma pathogenesis model . As showSince physical development and testis weight was altered in the asthmatic mice, we further sought to determine whether asthma could decrease sperm count and motility in the male mice. Intriguingly, results from hemocytometer measurements showed that both sperm count and motility in the asthma group were significantly lower than those in the control group . Consistent with the increased cleavage of caspase-3, the protein level of cleaved PARP was significantly upregulated in the asthmatic testis .Asthma may have a direct effect upon physical development in children. It is an important determinant of physical development particularly for young patients with severe asthma and without proper treatment . HoweverIn this study, it was shown that OVA-induced asthma has significant effects on the physical development of mice, including the body weight loss and decreased testis weight. These defects in physical development may directly affect the number of mature sperm cells in asthmatic testis. In addition, a more significant decrease in sperm motility and viability was also found, indicating that beyond the physical development, some causes may take part in the decreased spermatogenesis. Through DNA ladder assay and immunohistochemistry, OVA-induced apoptosis was confirmed in the testis of asthmatic mice as compared with the control group. Further mechanistic studies validated the observation and demonstrated that the caspase apoptosis pathway was activated significantly in the testis of OVA-treated mice. In fact, it has been estimated that up to 75% of the hypothetical sperm number is lost due to the programmed cell death , while dPreviously, it has been demonstrated that HIF-1α expression is potentiated in the lung of asthma mouse models, as well as in BALF cells from asthmatic patients . Here, wAs downstream events of apoptosis, we determined the enzymatic activity of caspase-9 and caspase-3/7, together with the cleavage of caspase-3 and PARP. These results demonstrate that the caspase apoptosis pathway is markedly activated in asthmatic testis compared with the control group, which might be the reason for decreased sperm viability and mobility. Previously, it was reported that apoptosis is not mediated exclusively by caspases. Among them, Apoptosis-inducing factor (AIF) was the first identified caspase-independent cell death effector which may suffice to induce cell death. AIF may be released from the mitochondrial intermembrane space along with cytochrome c and translocate to the nucleus, where it promotes DNA fragmentation, probably through endonuclease G (EndoG) . InteresAs revealed by the immunohistochemistry , a signiConclusively, the present study demonstrates that asthma could induce apoptosis in testis tissue possibly through the mitochondrial pathway, in which caspase-9, caspase-3 and PARP are significantly activated. As a result, apoptosis in asthmatic testis may ultimately lead to the infertility problems in male. To our knowledge, this is the first report that asthma could lead to the enhanced activation of mitochondrial apoptosis signaling pathway in the testis of mice.S1 Fig(A) control group. (B) asthma group.(TIF)Click here for additional data file.


In recent history, some dietary recommendations have treated dairy fat as an unnecessary source of calories and saturated fat in the human diet. These assumptions, however, have recently been brought into question by current research on regular fat dairy products and human health. In an effort to disseminate, explore and discuss the state of the science on the relationship between regular fat dairy products and health, symposia were programmed by dairy industry organizations in Europe and North America at The Eurofed Lipids Congress (2014) in France, The Dairy Nutrition Annual Symposium (2014) in Canada, The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology (2015) in the United States, and The Federation of European Nutrition Societies (2015) in Germany. This synopsis of these symposia describes the complexity of dairy fat and the effects regular-fat dairy foods have on human health. The emerging scientific evidence indicates that the consumption of regular fat dairy foods is not associated with an increased risk of cardiovascular disease and inversely associated with weight gain and the risk of obesity. Dairy foods, including regular-fat milk, cheese and yogurt, can be important components of an overall healthy dietary pattern. Systematic examination of the effects of dietary patterns that include regular-fat milk, cheese and yogurt on human health is warranted. Summary of lecture presented by Dr. Mario Kratz at The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology, 2015 (USA) [Over the last three decades, dietary guidelines have concluded that dairy fat is an unnecessary source of calories and saturated fat in the human diet ,3. The atrans-fatty acids and branched-chain fatty acids, all of which have biological significance [These assumptions, however, have recently been questioned by current research on regular fat dairy products and human health. The first assumption, that dairy foods are an unnecessary source of saturated fat, does not take into account that dairy fat contains nearly 400 different fatty acids, of which saturated fatty acids are a part, constituting the largest proportion of total dairy fat . Dairy fificance . The secificance . Plasma ificance . Furtherificance . The thiificance .In an effort to disseminate, explore and discuss the state of the science on the relationship between regular fat dairy products and human health, symposia were programmed by dairy industry organizations in Europe and North America at The Eurofed Lipids Congress (2014) in France, The Dairy Nutrition Annual Symposium (2014) in Canada, The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology (2015) in the United States, and The Federation of European Nutrition Societies (2015) in Germany. This article is a synopsis of what was presented at those meetings regarding regular-fat dairy products and human health.Summary of lecture presented by Dr. J. Thomas Brenna at The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology, 2015 (USA) .n-6 and n-3 PUFA, because replacement with pure n-6 polyunsaturated fat may increase risk [Current dietary recommendations indicate there is strong evidence that replacing saturated fat with unsaturated fats, particularly polyunsaturated fat, reduces plasma LDL-cholesterol and cardiovascular disease risk . Accordiase risk . There iase risk . The repase risk .trans-fatty acids, as well as branched-chain fatty acids [Humans eat complex foods containing a mixture of fatty acids, and thus dietary recommendations can be confusing to consumers, particularly when it comes to dairy foods . The major source of calories from cow’s milk is from saturated fat, but cow’s milk also contains monounsaturated fatty acids (MUFA) and PUFA, naturally occurring ty acids . Furtherty acids . TherefoThe numerous fatty acids in dairy fat are often grouped by categories of chemical classification, with SFA being the primary component of dairy fat . PalmitiSaturated fat is known to raise total cholesterol, including HDL-cholesterol and LDL-cholesterol. Higher HDL-cholesterol is thought to reduce cardiovascular disease risk by reverse cholesterol transport, the inhibition of LDL-cholesterol oxidation, and thus prevention of subsequent inflammatory pathways .trans-fatty acids [trans-11 18:1 (vaccenic acid) is the most abundant trans-fatty acid in retail milk [trans-fatty acids, such as those present in partially hydrogenated oils, the consumption of naturally-occurring trans-fatty acids present in ruminant milk have not been associated with increased risk of cardiovascular disease when consumed in typical amounts [cis-9, trans-11 18:2), a fatty acid that has been associated with antiatherogenic and anticarcinogenic effects in vitro and in vivo [trans-fatty acids on risk factors for cardiovascular disease in healthy adults, high levels of cis-9, trans-11 18:2  lowered triacylglycerol and had no effect on other lipoprotein risk factors [trans-fatty acids from any source, industrially- or ruminally-produced, including vaccenic acid, adversely affected plasma LDL and HDL cholesterol, intake of ruminally-produced trans-fatty acids, such as vaccenic acid, in moderate amounts that exceed current normal intake had neutral effects on plasma lipids and other cardiovascular disease risk factors [Dairy fat also contains several ty acids . At apprail milk . Unlike  amounts ,13. Conj in vivo ,15, thou in vivo ,17. A cl factors . Results factors .Another class of fatty acids present in dairy fat, the bioactivity of which has not been thoroughly investigated, is branched-chain fatty acids. Branched-chain fatty acids are primarily saturated fatty acids with one or more methyl groups. They make up about 2% of the total fatty acids in cow’s milk . The mosw/w) branched-chain fatty acids [Branched-chain fatty acids increase membrane fluidity, and are resistant to oxidation . In factty acids . The braty acids ,24. The Cow’s milk products, such as milk, cheese and yogurt, are major sources of fat in the American diet, with the intake of ruminant-derived branched-chain fatty acids being approximately 220 mg/day from dairy foods . This isThis brief survey of the bioactive fatty acids present in dairy fat highlights the complexity of the role of regular fat dairy in human health. To understand human disease risk, however, it is prudent to examine the effect from actual foods consumed, such as regular-fat dairy foods, including milk, cheese and yogurt, rather than individual fatty acids, on human health outcomes.Summary of lecture presented by Moises Torres-Gonzalez at The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology, 2015 (USA) .Since the discovery of correlations between dietary fat, serum cholesterol and cardiovascular disease, scientists have been discouraging the consumption of nutrient-dense foods that contain saturated fat and/or cholesterol, such as regular-fat milk, cheese and yogurt . The assAdditionally, despite clinical practice guidelines based on targeting a reduction in plasma LDL-cholesterol, still considered the main biomarker for CVD risk reduction , advanceFurther, the functionality of HDL-cholesterol affects its cholesterol efflux capacity, highlighting that concentration alone is not the best indicator of risk . Raised Another important aspect of cardiovascular disease is a presence of low-grade chronic inflammation . InflammCurrent dietary recommendations for saturated fat are based on targeting a reduction in plasma LDL-cholesterol, with the consumption of saturated fat recommended to be no more than 7%–10% of calories ,48,49. AEvidence from large randomized clinical trials shows that reducing the percentage of calories from saturated fat to less than or equal to 10% of total calories does not necessarily improve plasma biomarkers for cardiovascular disease, because these biomarkers do not account for the complexity of cholesterol metabolism . For exaA Mediterranean dietary pattern has been recommended as a heart healthy diet . A randoad libitum feeding and lacked detail regarding total diet, as well as detailed information regarding the inclusion criteria, physical activity and health status of participants. A randomized controlled trial, however, that tested whole vs. skim milk was since published. This study tested the diet recommended by the U.S. American Heart Association with either whole or skim milk included. Whereas total, LDL-cholesterol and ApoB levels were lower after the skim milk period compared to whole milk, there were no differences in HDL-cholesterol, non-HDL to HDL-cholesterol ratio, ApoAI and TAG between the two diets [Few studies have evaluated regular-fat dairy foods specifically in relation to plasma biomarkers for cardiovascular disease. One study found that adding three servings of regular fat yogurt to the habitual diet decreased serum cholesterol, thus suggesting a potential hypocholesterolemic effect by yogurt consumption . A studywo diets . Anotherwo diets .More research is needed to better understand the effects of regular fat dairy foods on markers of inflammation. It is important that future research recognize that cardiovascular disease is a multifactorial disease that cannot be defined by a single or narrow set of biomarkers. Reporting the effect of dairy foods on markers of biomarkers of inflammation, as well as on lipoprotein particle size and quantity may improve the capability of assessing the effects of dairy foods on cardiovascular risk.Summary of lecture presented by Dariush Mozaffarian at The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology, 2015 (USA) . The folA systematic review of the observational literature found that although data were mixed, the majority of studies indicated regular fat dairy consumption was not associated with cardiovascular events . WhereasA body of literature demonstrates that foods’ effects on obesity include complex influences such as on satiety, hormonal responses, hepatic de novo lipogenesis, reward and craving, gut flora responses, and metabolic expenditure that ultimately all calories are not created equal ,67,68,69Weight loss can be monitored over short-term dietary interventions, but the prevention of weight gain must be monitored over the long term. This is because the average adult gains only about one pound per year. An analysis of three large cohort studies that examined diet and weight change every four years among over 100,000 subjects that participated in the Nurses’ Health Studies over 24 years indicated that certain foods were associated with weight gain and others with weight loss . Cheese The evidence that regular-fat dairy foods do not contribute to weight gain is not restricted to observational data. A meta-analysis of randomized controlled trials of dairy consumption and weight change in adults indicated that total dairy consumption was associated with weight loss . The samtrans-palmitoleic acid , a biomarker of dairy fat intake, was associated with 50%–60% less incidence of type 2 diabetes in an analysis from four large prospective cohorts [Obesity is an important cause of type 2 diabetes . The inv cohorts . The assSummary of lecture presented by Arne Astrup at The Federation of European Nutrition Societies 12th European Nutrition Conference, 2015 (Germany) .trans-fatty acids, such as those present in meat and dairy foods [Currently, the World Heart Federation warns consumers to beware of “bad fats”, saturated and ry foods . This isry foods ,84. A dory foods . A studyry foods .Despite its contribution of total and saturated fat to the diet, regular-fat dairy has not been shown to negatively impact human health . CurrentThe evidence from both observational and clinical research indicates that cheese, when consumed as part of a healthy diet, is not linked to increased risk of obesity, metabolic syndrome, type 2 diabetes or cardiovascular disease. Further, cheese does not exert effects on blood lipids and blood pressure as has been traditionally predicted by its sodium and saturated fat content. Thus, the effects of cheese on metabolic health highlight the pitfalls of basing dietary guidance on nutrients, rather than foods.Summary of lecture presented by Bernadette Delplanque at European Federation for the Science an Technology of Lipids 12th Euro Fed Lipids Congress (France) .Up until the 20th century, regular-fat cow’s milk was used to manufacture formulas to feed infants. As formulas evolved to meet specific needs of essential fatty acids, they became increasingly based on cow-milk proteins and blends of pure vegetable oils. Whereas the need for essential fatty acids are met, numerous other components of human milk, such as cholesterol fatty acids, TAG and globule fat structures are not found in pure vegetable blends. In this way, regular-fat cow’s milk is closer in fat composition to human breast milk than vegetable blends. The fatty acids and components missing from pure vegetable blends during the perinatal period may be crucial for later development and healthy adulthood .n-6 PUFA and about 0.8%–3.6% n-3 PUFA [Recommendations for infant formulas have been established on the premise that human breast milk is the “gold standard” that formulas should mimic. The content of lipids in human milk varies by geographical location and during lactation (3–4 g/100 g), and is mostly represented as TAG (95%). Human milk fat contains 34%–47% of saturated fatty acids , about 31%–43% monounsaturated fatty acids (oleic acid: 26%–36%) and about 12%–26% n-3 PUFA .alpha-linolenic acid (ALA) are essential fatty acids present in human breast milk. ALA is a precursor for long-chain fatty acids, such as docosahexaenoic acid (DHA), which are necessary for proper brain development in infants. For decades, the ideal amount and proportion of LA and ALA has been debated, and the recommendations have changed accordingly. The LA and ALA content in breast milk depends on the mother’s diet and thus varies widely between countries; for LA, 10% to 24% of fatty acids and for ALA, 0.6% to 1.9% of fatty acids [Linoleic acid (LA) and ty acids . Over thty acids  reportedty acids ,102. Thety acids . Researcty acids ,105,106.Recently, infant formulas based on blends of vegetable oils mimicked quite well the 20th century composition of human breast milk in terms of essential fatty acids. Not all, however, contained cholesterol, certain short- and medium-chain fatty acids or the proper sn2-position of palmitic acid on TAG. Some of these components could be provided by the addition of specific oils or products .trans- and branched-chain fatty acids for a total of approximately 20% of milk fatty acid content. Functional purposes of the fatty acids present in cow’s milk have also been demonstrated [Cow’s milk fat naturally presents fewer differences from human breast milk than vegetable blends. Cow’s milk fat offers a better representation of sn2-position of palmitic acid, has similar content of cholesterol, short- and medium-chain fatty acids (C6:0 to C12:0), all nstrated ; for exanstrated . The prenstrated . These fnstrated .n-6) over the last 50–60 years.The most striking difference between breast milk and cow’s milk is the content of LA, which is approximately 10 times less in cow’s milk , while ALA is quite similar (0.7%–2% vs. 0.4%–08%). Thus, the drastic differences in the LA/ALA ratio from 10 in breast milk to less than 3 in cow’s milk could be attributed to changes in the human diet that has been observed (enriched in n-6 and n-3 fatty acids (arachidonic acid (ARA) and DHA, respectively). Breast milk contains very low levels of DHA and ARA  and cow’s milk contains approximately 5 to 10 times less [Currently, all manufactured infant formulas are made of blends of vegetable oils, which contain average LA and ALA content (12%–15% and 1.5% to 2.5% respectively). The specific amounts depend upon the type and source of fats used to make the formula. The LA/ALA ratio in infant formulas varies from 5 to 15. Some formulas contain added long-chain mes less .Adequate DHA is crucial from three months before delivery to five years after birth, for rapid growth and differentiation of the brain. The addition of DHA to infant formula has not been consistently shown to have benefits in visual, neural or growth outcomes . Some stThe effects of reintroducing dairy fat into infant formulas on blood and brain levels of DHA recently has been studied in young rats. The rat is an appropriate animal model for these nutritional studies and has been used previously to establish recommendations for infant nutrition ,113. Blen-3 (DHA) levels; Second, the dairy-fat-based diet  enriched with 2.3% ALA was even more efficient and this could be attributed to both the increased level of dietary ALA and the concomitant decrease in the LA/ALA ratio (from 10 to 5). A lower LA/ALA ratio has been recognized as an important factor driving the bioconversion of ALA into DHA because of the competition between the parent n-3 and n-6 fatty acids for desaturation and elongation pathways; Third, the dairy-fat-based diet containing pure dairy fat  was as efficient as the rapeseed diet , and comparable to the 2.3% ALA dairy/vegetable blend diet, all of them containing a very low LA/ALA ratio (3 to 5). Delta6-desaturase could be involved in this process, and its role is crucial to explain these last results because ALA is the precursor of DHA but also its competitor for the last step of the delta6-desaturase pathway, and is regulated by substrate levels [n-3 and secondarily limit delta6-desaturase activity in the second control point for DHA conversion, as observed for rapeseed oil, which could act as an excess precursor as shown previously [n-6 and n-3 precursors. Pure dairy fat with only 0.8% of ALA and a LA/ALA ratio around 3 is quite sufficient to provide required brain levels of DHA. This study also demonstrated that despite very low levels of PUFA  and a favorable LA/ALA ratio similar to rapeseed oil (maximum 3/1), pure dairy fat was able to provide adequate bioconversion of ALA to long-chain n-3 and DHA necessary for the brain of young animals.This study had three main findings. First, a dairy-fat-based diet  was more efficient than a pure vegetable oil blend with the same amount of ALA (1.5%) and LA/ALA ratio (10), to increase brain DHA. Specific and complex components of dairy fat, such as the short- and medium-chain fatty acids which are highly oxidizable after absorption ,119, maye levels . An exceeviously ,123. Resn-3 status than those fed formulas enriched with LA-rich vegetable oils. The use of fats that are low in PUFA such as dairy may confer some metabolic advantages in that they allow better endogenous conversion of ALA to DHA. Consequently, the use of dairy fat in infant formulas should be reconsidered, as well as the recommended absolute amount of polyunsaturated LA and ALA.Together, these observations demonstrate that brain DHA levels could be substantially improved by dairy fat based-diets. These results are similar to the results obtained in studies showing that infants fed formulas based on dairy fats  had bettSummary of lectures presented by Dr. Dariush Mozaffarian at The American Society for Nutrition Annual Meeting held in conjunction with Experimental Biology, 2015 (USA) and Dr. Benoît Lamarche at the Dairy Nutrition Annual Symposium, 2014 (Canada) ,125. TheThe state of the science indicates that consuming lower fat dairy foods does not contribute to improved body composition and metabolic health when compared to regular fat dairy foods ,89. RathWhereas more research on the relationship between dairy fat intake and cardiovascular disease and type 2 diabetes risk is warranted ,126, the


Better methods for purifying human or mouse acinar cells without the need for genetic modification are needed. Such techniques would be advantageous for the specific study of certain mechanisms, such as acinar-to-beta-cell reprogramming and pancreatitis. Ulex Europaeus Agglutinin I (UEA-I) lectin has been used to label and isolate acinar cells from the pancreas. However, the purity of the UEA-I-positive cell fraction has not been fully evaluated. Here, we screened 20 widely used lectins for their binding specificity for major pancreatic cell types, and found that UEA-I and Peanut agglutinin (PNA) have a specific affinity for acinar cells in the mouse pancreas, with minimal affinity for other major pancreatic cell types including endocrine cells, duct cells and endothelial cells. Moreover, PNA-purified acinar cells were less contaminated with mesenchymal and inflammatory cells, compared to UEA-I purified acinar cells. Thus, UEA-I and PNA appear to be excellent lectins for pancreatic acinar cell purification. PNA may be a better choice in situations where mesenchymal cells or inflammatory cells are significantly increased in the pancreas, such as type 1 diabetes, pancreatitis and pancreatic cancer. This in vitro testing requires an optimal technique for purifying acinar cells from the pancreas82122232526282930In order to optimize acinar-to-beta cell reprogramming, Tomato  mouse model to isolate acinar cells based on red fluorescence by flow cytometry, and have shown that the purified acinar cells lack contaminating non-acinar pancreatic cells, such as synaptophysin (SYN)-positive endocrine cells, CD31-positive endothelial cells, Dolichos biflorus agglutinin (DBA)-positive duct cells, vimentin-positive mesenchymal cells and CD45-positive inflammatory cells3233Isolation of acinar cells using genetically modified mice in which a fluorescent reporter is expressed under an acinar-cell-specific promoter has its advantages and disadvantages. For example, we have previously used a tamoxifen-treated elastase-CreERT; ROSA26Earlier studies have utilized lectin affinity to acinar cells to purify acinar cells by flow cytometry21All the mouse experimental methods in the current study were approved by the Animal Research and Care Committee at the Children’s Hospital of Pittsburgh and the University of Pittsburgh IACUC (Protocol number: 14031989). All the experiments have been carried out in accordance with the guidelines from the research committee at the Children’s Hospital of Pittsburgh and the University of Pittsburgh.Tomato  has been described before3233C57BL/6 mice and non-obese diabetes (NOD) mice were purchased from Jackson Lab . Bacterial-artificial-chromosome (BAC) transgenic elastase-CreERT; ROSA26Pancreatic duct perfusion and subsequent complete digestion of the pancreas were performed with 0.25 mg/ml collagenase  for 50 minutes to obtain a single cell population, as has been described before834832−ΔΔCt method. Values for genes were first normalized against CypA, the housekeeping gene control, and then compared to the complete pancreatic digests, without sorting, as an experimental control.RNA was extracted from digested pancreatic cells w/o FAC sorting using RNeasy  and quantified with Nanodrop1000  according to the manufacturer’s instructions, followed by cDNA synthesis (Qiagen). Primers were purchased from Qiagen for cyclophilinA , amylase (QT00179242), CK19 (QT00156667), CD31 (QT01052044), vimentin (QT00159670) and CD45 (QT00139405). RT-qPCR reactions were performed in duplicates with QuantiTect SYBR Green PCR Kit (Qiagen) using a LightCycler 1.5 Instrument . Specificity of the amplified products was determined by melting point analysis. Quantification for each gene of interest was performed with the 2Immunostaining and Western Blot analysis were performed as described before32333637832All values are depicted as mean ± standard error of the mean. Five mice were analyzed in each experimental group. All data were statistically analyzed using one-way ANOVA with a Bonferoni correction, followed by Fisher’s Exact Test for comparison between two groups, with a GraphPad Prism 6.0 .UEA-I lectin has been used to isolate acinar cells from the pancreas. Nevertheless, the exact purity of the acinar cell fraction has not been thoroughly evaluated. Here, we aimed to compare the binding affinity and specificity of different lectins for acinar cells in order to ascertain the optimal lectin for this purpose. First, we stained the mouse pancreas with 20 widely used FITC-conjugated lectins , and fouTomato  mouse pancreas , and inflammatory cells (positive for CD45). However, these two minor populations can be significantly enriched in some pathological processes. For example, during pancreatitis and pancreatic fibrotic diseases, the number of mesenchymal cells dramatically increases, whereas during pancreatic inflammation (e.g. pancreatitis or autoimmune diabetes), the number of inflammatory cells present in the pancreas may be significantly increased. In these pathologic situations, the purity of acinar cells that are isolated based on lectin affinity could be significantly affected by the affinity of lectin to these other cell populations. Thus, we purified acinar cells from either UEA-I-labeled C57BL/6 mouse pancreas  based onpancreas  based onpancreas . The purThe purity of sorted acinar cells based on UEA-I or PNA binding, or by tomato fluorescence in Ela-Cre; TOM mice was compared to the unlabeled dissociated pancreatic cells from C57BL/6 mice by examination of mRNA levels of specific cell marker genes, including amylase (for acinar cells), SYN (for endocrine cells), CK19 (for duct cells), CD31 , vimentin , and CD45 (for inflammatory cells).For each cell type population, the fold change for that cell type-specific gene (e.g. CK19 for duct cells) is graphed, compared to the transcript level for that gene in the total pancreas. Thus, for example the CK19 value for PNA-sorted pancreas is roughly 17% of the CK19 for the whole pancreas. If we estimate that 2% of the whole pancreas typically contains ducts, then only 17% of those 2%, or 0.34% of the cells in the PNA-sorted preparation are contaminating duct cells.We found that the purity of acinar cells sorted by PNA was equivalent to that of Ela-Cre; TOM mice , suggestTo confirm the above findings, we used UEA-I  and PNA in vivo, and in vitro, through either gene manipulations or application of growth hormone cocktails18192021in vitro, which in turn is dependent on optimal purity of the cell population.The generation of functional pancreatic beta-cells to compensate for the short supply of donor human islets has long been regarded as a potential solution for the treatment of diabetes40Very pure acinar cell populations can be isolated from genetically modified mice in which a fluorescent reporter is expressed under an acinar-cell-specific promoter, e.g. previously described Ela-Cre; TOM mouse3343444546As discussed before, early studies used lectin binding to acinar cells to purify them by flow cytometry21UEA-I was used in previous studies for purifying acinar cells21Here we focused on lectin-purification of acinar cells with regard to possible applications to diabetes research. However, purification of acinar cells may be an important adjunct to studies in pancreatitis and pancreatic carcinoma, since acinar cells may be involved in these diseases. Our study provides evidence for the preferred use of PNA rather than UEA-I to purify primary acinar cells from mouse pancreas, especially in diseases involving inflammation and tissue remodeling.How to cite this article: Xiao, X. et al. PNA lectin for purifying mouse acinar cells from the inflamed pancreas. Sci. Rep.6, 21127; doi: 10.1038/srep21127 (2016).


Successfully treated HIV-infected patients may still have an increased risk for cardiovascular morbidity and mortality, which might be related not only to traditional risks, but also to inflammation and dyslipidemia induced by HIV and/or antiretroviral therapy , 2. We eSubjects were selected among consecutive HIV-infected males ≥18 years of age with an undetectable viral load (<50 copies/mL of HIV1-RNA), seen at the University Hospital for Infectious Diseases, Zagreb, Croatia, in the period from January 2012 to March 2013. Plasma inflammatory biomarkers  >75th percentile were considered elevated and an IBS was constructed as the presence of zero, one, two, or three or more elevated biomarkers. Correlations between the IBS and lipid parameters were examined using Spearman's Rho and by ordered logistic regression proportional odds model to estimate the odds of more elevated (>75th percentile) biomarkers.181 male patients were included into the study, the median age was 46.7  years and the median current CD4 cell count was 553.0  per microliter. The patients were mainly treated with two nucleoside reverse transcriptase inhibitor (NRTI) plus one non-NRTI (NNRTI)  or two NRTI plus lopinavir . There was a significant correlation between the IBS and serum cholesterol , triglycerides  and cholesterol/HDL-cholesterol ratio . In the multivariable model a one unit increase in cholesterol/HDL-cholesterol ratio was associated with a 1.72-fold  increased odds of having a greater IBS. One unit increase (mmol/L) of cholesterol and triglycerides was associated with a 1.41-fold  and 1.37-fold  increased odds of having a greater IBS, respectively.Our study suggests that in virologically suppressed patients there is a significant association between markers of inflammation and serum levels of cholesterol and triglycerides as well as the cholesterol/HDL-cholesterol ratio.


Accessory navicular can become symptomatic in childhood, and in some cases, the condition is associated with progressive flattening of the longitudinal arch. Moreover, some severe, rigid flatfoot deformities are associated with an accessory navicular. We investigated the results of concomitant calcaneo-cuboid-cuneiform osteotomies (triple C) and the modified Kidner procedure for severe flatfoot associated with a symptomatic accessory navicular in children and adolescents.Twenty-one feet of 13 patients  with severe flatfoot associated with a symptomatic accessory navicular who were treated with concomitant triple C and the modified Kidner procedure were evaluated based on clinical and radiographic examinations preoperatively and at a mean follow-up of 22.4 months . We measured 12 variables on the anteroposterior (AP) and lateral weight-bearing radiographs, and we used the American College of Foot and Ankle Surgeons (ACFAS) score for clinical assessment.p < 0.001) in eight of the 12 radiographic measurements: the AP talo-first metatarsal (MTT) angle, AP talo-navicular coverage angle, AP talo-calcaneal angle, lateral talo-first MTT angle, calcaneal pitch, lateral talo-calcaneal angle, lateral talo-horizontal angle, and naviculo-cuboid overlap. Average ACFAS scores were significantly improved at the time of the last follow-up (p < 0.001). The only complication was overcorrection of the hindfoot in one patient.We found significant improvements (Concomitant triple C and the modified Kidner procedure result in favorable radiographic and clinical outcomes in the treatment of severe flatfoot associated with a symptomatic accessory navicular in children and adolescents. An accessory navicular is a supernumerary ossicle that occurs within the tibialis posterior tendon (TPT) at its navicular insertion. Symptoms develop with recurrent stress at the synchondrosis between the accessory bone and the medial navicular tuberosity secondary to a pull of the TPT or trauma . Kidner As with most orthopedic conditions, the first line of treatment for an accessory navicular is nonoperative. Operative intervention is indicated for patients who are recalcitrant to nonoperative treatment and have persistent pain and dysfunction. Current operative treatments include simple excision of the accessory bone from within the TPT and the modified Kidner procedure, which involves excision with takedown and reattachment of the TPT (with or without advancement) -7. TheseIn the current study, we assessed the outcomes of combined triple C and the modified Kidner procedure for severe, symptomatic flatfoot associated with an accessory navicular in children and adolescents. We hypothesized that these procedures would lead to improvement of pain, function, and correction of the flatfoot deformity.This study was approved by the institutional review board at our institution. The presence of an accessory navicular was confirmed on plain radiographs, and the diagnosis of flatfoot deformity was based on a poor formation of the arch and a valgus position of the heel during weight bearing. The inclusion criteria were (1) consecutive patients who underwent a combined triple C and modified Kidner procedure for severe flatfoot deformity associated with an accessory navicular between 2006 and 2012, (2) patients who had preoperative and postoperative weight-bearing anteroposterior (AP) and lateral foot radiographs, and (3) patients with a minimum follow-up of 1 year. Patients with severe deformity were those who could not reconstitute the arch and who had a varus position of the heel while on tiptoes. The exclusion criteria were underlying neurological or syndromic conditions, tarsal coalition, or inadequate radiographs for measurement.During the study period, 66 patients (70 feet) were diagnosed with a symptomatic accessory navicular and underwent operative treatment. Of those patients, 55 had concomitant flatfoot deformity. Twenty-four feet had severe deformity and underwent a combined triple C and modified Kidner procedure. Three feet were lost to follow-up. The other 31 feet were treated with the modified Kidner procedure only. Twenty-one feet of 13 patients were available for follow-up and were included in this study. There were nine boys and four girls with a mean age at the time of surgery of 12.7  years. The mean follow-up was 22.4  months. All were type II accessory navicular. All procedures were performed by a single surgeon (KJR).The surgical technique for triple C was similar to that reported by Rathjen and Mubarak . The latThe modified Kidner procedure was then performed through another medial incision. The ossicle was identified and excised together with the entire synchondrosis and any residual navicular prominence. The tendon was completely detached from the TPT insertion site. The plantar medial tuberosity of the navicular was decorticated. Positioning of the anchor was confirmed under fluoroscopy to ensure that the optimal reattachment point was used and that the navicular-cuneiform joint was not violated. The TPT was advanced and attached to the navicular at neutral resting tension, utilizing the two no. 2 braided nonabsorbable sutures attached to the anchor  Figure c, d. WitPreoperative and final postoperative follow-up weight-bearing AP and lateral radiographs of the foot were available for all cases. The standing AP radiographs were used to calculate the following four measurements: talo-calcaneal angle, talo-first metatarsal (MTT) angle, talo-fifth MTT angle, and talo-navicular coverage. The standing lateral foot radiographs were used to calculate the following eight measurements: talo-calcaneal angle, talo-first MTT angle, talo-horizontal angle, calcaneal pitch, calcaneo-fifth MTT angle, first-fifth MTT angle, naviculo-cuboid overlap, and medial-lateral column ratio. All radiographic measurements were performed as in Davids et al.  and MoraThree orthopedic surgeons  assessed the interobserver reliability of the radiographic measurements. Each examiner took the measurements independently, without knowledge of the patients’ clinical information and the other orthopedic surgeons’ measurements. All measurements were collected by a research assistant (LZ) who did not otherwise participate in this study.Clinical outcomes were assessed with the American College of Foot and Ankle Surgeons (ACFAS) scoring system preoperatively and at the time of final follow-up. The flatfoot module of the ACFAS score includes both subjective and objective assessment . This syt-tests were used for the statistical analysis. All analyses were performed with SPSS version 21.0 for Windows . p values <0.05 were considered statistically significant.Paired With the exception of the medial-lateral column ratio, all radiographic measurements showed good to excellent interobserver reliability Table .Table 1p = 0.015). However, this measurement was not useful because interobserver reliability was 0.58 , 0.14–0.82). The average ACFAS score improved significantly from 26.1 (±6.8) preoperatively to 44.8 (±5.7) postoperatively (p < 0.001) (Table Correction of preoperative deformity was achieved to the normal range in 78% of patients for AP talo-first MTT angle, 69% for AP talo-navicular coverage angle, 91% for talo-calcaneal angle, 46% for AP talo-fifth MTT angle, 70% for lateral talus-first MTT angle, 72% for calcaneal pitch angle, 67% for lateral talo-calcaneal angle, 74% for lateral talo-horizontal angle, 63% for lateral naviculo-cuboid overlap, 45% for lateral calcaneo-fifth MTT angle, and 47% for lateral first-fifth MTT angle. A comparison of the preoperative and final follow-up weight-bearing radiographs showed significant improvement in eight of the 12 radiographic measurements: AP talo-first MTT angle, AP talo-navicular coverage angle, AP talo-calcaneal angle, lateral talo-first MTT, calcaneal pitch, lateral talo-calcaneal angle, lateral talo-horizontal angle, and lateral naviculo-cuboid overlap. We also found marginally significant improvements in three of the 12 radiographic measurements: AP talo-fifth MTT angle, lateral calcaneo-fifth MTT angle, and lateral first-fifth MTT angle (Table On physical examination, we observed a neutral arch in 17 cases and a flat arch in three cases. There was one varus alignment of the hindfoot due to overcorrection. All forefeet were in a neutral position. There were no wound complications or infections. All patients rated their results as good or excellent. No patients had developed any recurrence of flatfoot deformity either clinically or radiographically at the latest follow-up Figure .Figure 2The overall effect of an accessory navicular on foot mechanics is a source of controversy, particularly regarding the association with flatfoot deformity. Flatfoot deformity is present in approximately 50% of patients who become symptomatic, although its role in the development of symptomatic accessory navicular remains unclear . After tThe initial management of a symptomatic accessory navicular is always conservative. When symptoms become intractable despite adequate conservative treatment, surgical intervention may be indicated. Type II accessory navicular is most likely to require surgical management. The classic Kidner procedure requires excision of the ossicle and rerouting of the TPT to the plantar aspect of the navicular in order to restore the TPT’s ability to support the medial arch. Several investigators have questioned the need for detachment of the TPT, opting instead for excision of the ossicle through a split in the tendon that is subsequently repaired ,15-17. WIn cases that involve a more severe, yet flexible, deformity, additional correction can be achieved in conjunction with a Kidner-type procedure with the addition of a subtalar arthroereisis . The proMany investigators have noted that radiographic measurements are related to clinical outcomes because they provide reliable information on the relationships of different parts of the hindfoot ,23. ReceOur study had some limitations. First, we included only a small number of patients with a short follow-up period and no control group. Second, the inclusion of unilateral and bilateral cases in the same group could violate the principle of statistical independence. Nevertheless, the study population was homogeneous in terms of the etiology of the condition. Our results have demonstrated that combining triple C with the modified Kidner procedure can correct both the painful prominence and flatfoot deformity. These procedures lead to an appropriate length of the otherwise healthy TPT, and the TPT is able to support the arch, as in a person with normal arch mechanics. Therefore, triple C combined with the modified Kidner procedure can be an effective treatment option for severe flatfoot deformity associated with a symptomatic accessory navicular in children and adolescents.Triple C combined with the modified Kidner procedure can be an effective treatment option for severe flatfoot deformity associated with a symptomatic accessory navicular in children and adolescents.


Internet-based cognitive-behavioral therapy (iCBT) for social anxiety disorder has been found effective, as attested by independently conducted randomized controlled trials in four languages. The study aim is to test the efficacy of an iCBT program in a culture where it was not tested before (i.e. Romania).n = 76) were recruited, screened and randomized to either a nine-week guided iCBT or a wait-list control group in April and May 2012. Self-report measures were collected before (April 2012) and after the intervention (July 2012), as well as six months later (January 2013). Although social anxiety was assessed with multiple measures, the Liebowitz Social Anxiety Scale - Self Report version (LSAS-SR) and Social Phobia Inventory (SPIN) were used as the primary outcome measures.Participants (d = 1.19 for LSAS-SR and d = 1.27 for SPIN). Recovery rates show that 36.8% (n = 14) in the treatment group score below the SPIN clinical cut-off compared to only 2.6% (n = 1) in the wait-list control group. Post-intervention clinical interviews also revealed that 34.2% (n = 13) of the treatment group was completely recovered (full remission) while additionally 36.8% (n = 14) retained some social anxiety symptoms . However, an important study limitation is that post-intervention interviewers were not blinded to the study conditions. The program also effectively reduced depression and dysfunctional thinking . Moreover, the iCBT intervention appears to have a long-term impact for participants’ functioning, as the treatment gains were maintained six months later.A significant difference with a large between-group effect size in favor of iCBT was found (Cohen´s Internet-delivered interventions display a high potential to quickly and widely disseminate effective evidence-based programs around the world. This study provides support for guided iCBT as a promising treatment approach in Romania.NCT01557894ClinicalTrials.gov  Social anxiety disorder (SAD), also referred to as social phobia , is defiSAD varies in severity and a distinction is commonly made between limited/non-generalized  and generalized SAD . The disThere are a range of psychological and pharmacological treatments for SAD .The mostCognitive models of SAD emphasize the importance of dysfunctional beliefs in generating the disorder, by transforming innocuous social cues into threats , 18. ThuDespite the availability of effective interventions, only a minority of individuals suffering from SAD seek and receive appropriate treatment , 21. ManIn an attempt to increase the accessibility of adequate, cost-effective treatments, researchers have focused on developing internet-based cognitive-behavioral therapy interventions (iCBT). In most studies, a guided self-help approach has been used, where a web-based presented program is combined with minimal, but regular therapist contact by e-mail or phone . Aside fd = .86) and moderate effects on quality of life (d = .53), and comorbid anxiety and depression (d = .43). Considering these encouraging results, it is important that iCBT for SAD are further examined in randomized clinical trials, with major theoretical and practical objectives.The first iCBT trials for SAD were conducted by Andersson and colleagues using a pInternet Social Phobia (iSOFIE)-was originally written in Swedish[The first study objective was to investigate the efficacy of any CBT program in a culture where it has not been tested before. Our intervention—called n Swedish, being sConsistent with the evidence-based approach that encourages the effort to test the same program in various contexts and cultures, we investigated the efficacy of an intervention program in a different culture, filling this gap in the literature. Besides enlarging the empirical support of an existing psychological service , the international availability of an empirically-supported intervention has the potential to alleviate the suffering of many socially anxious individuals in Romania .The second study objective was to look at modifications in appraisal/evaluative beliefs following this brief iCBT intervention. To date, only one study compared the rational-emotive behavior therapy (REBT) with the classical CBT approach and found them equally effective in reducing SAD symptoms. To our The protocol for this trial and supporting CONSORT checklist are available as supporting information see .This study was approvedby the Council of Scientific Research Ethics Comission of Babes-Bolyai University, Cluj-Napoca, Romania (Registration No 30273). Written informed consent was obtained from all participants by surface mail.We designed a single-center, parallel group randomized clinical trial (RCT), with equal randomization for the two conditions . Stratified randomization with diagnostic status as a factor  allowed the wait-list sub-clinical participants to serve as controls for their counterparts allocated to the iCBT condition.n = 291). Since the iSOFIE is a disorder-specific program, we only selected participants who could attain the maximum benefits from this intervention . After a rigorous check 76participants were recruited for the intervention (see SD = 8.04). The demographic characteristics for each of the two groups as well as for the total sample are summarized in Volunteer applicants from across the country were screened for the study  [Social Interaction and Anxiety Scale (SIAS)  [Liebowitz Social Anxiety Scale—Self Report version (LSAS-SR)  [Social Phobia Screening Questionnaire (SPSQ) [Structured Clinical Interview for DSM-IV-TR (SCID) [Beck Depression Inventory-II (BDI-II) [Screening Questionnaire of the SCID), (f) not currently receiving other forms of psychological treatment for SAD, (g) having access to a computer connected to the internet, (h) if on medication, the dose should be constant for at least 1 month, and participants should agree to keep the dosage unchanged for the whole duration of the study, and (i) having no diagnosis of psychoses or borderline personality disorder on the SCID. The accepted cut-off scores for the SAD measure according to the recommendations found for each instrument in their original article were used. However, because SPIN has the most reliable empirical support for its clinical cut-off, the recovery rates were estimated only for SPIN.Eligibility criteria for the study included: (a) being over 18 years old, (b) exceeding the cut-off score on ore ≥ 19), Social re ≥ 24) , and Liere ≥ 30) ,(c) fulfe (SPSQ) , (d) havR (SCID) , (e) pre(BDI-II) , and notTo be included in the study, participants were allowed to have prior history of treatment for SAD, but in the treatment group only five participants reported receiving treatment within the last five years (which speaks of the low rate of SAD treatment in Romania).The present intervention was adapted from a manual  previoushttps://www.iterapi.se/sites/isofie) was provided. Interested participants registered online, read the informed consent, and filled in the screening questionnaires over the internet. Those who fulfilled the minimal inclusion criteria  were contacted for a telephone interview. The interview was conducted within 12 days after the completion of the screening, and was based on the SCID. The purpose of this interview was to assess the applicant’s diagnostic status according to the DSM-IV criteria for SAD, and to briefly check for possible major problems undetected during screening. Sixty-six applicants (out of 121 who were contacted for the interview) meet the DSM-IV criteria for SAD. Ten additional applicants presented subclinical symptoms . In total, 76 applicants were included in the study as we predicted that they could benefit from this low-intensity psychosocial intervention. The flow of participants throughout the study is diagramed in The trial was briefly presented in various national and local newspapers , where a link to the project website (n = 38) or to the wait-list control condition (n = 38) via a computerized randomization procedure  consisted of a short telephone interview  and the same online questionnaires as the ones completed during the screening phase. In order to assess the satisfaction with the intervention, the iSOFIE participants were further invited to fill in a treatment satisfaction questionnaire. After the post-intervention week , participants were no longer able to communicate on the platform, but they were able to access all the modules for the next six months . Finally, control group participants started the iCBT intervention one week after the iSOFIE group completed the program.Liebowitz Social Anxiety Scale—Self-Report version (LSAS-SR) [The LSAS-SR)  presentsLSAS-SR)  and the LSAS-SR)  and adolLSAS-SR) .Social Phobia Inventory (SPIN) [The y (SPIN) , is a bry (SPIN) , 43and wy (SPIN) .Social Interaction and Anxiety Scale (SIAS) [Besides these primary outcome measures, social anxiety symptoms were assessed with two additional measures. The e (SIAS) , is a 20e (SIAS) .Social Phobia Screening Questionnaire (SPSQ)[The re (SPSQ), is a dire (SPSQ). SPSQ ware (SPSQ), 37, 38.Beck Depression Inventory-II (BDI-II) [The severity of depressive symptoms according to DSM-IV was measured with the (BDI-II) . Each it(BDI-II)  and it w(BDI-II) .Cognitive factors related to emotional problems were explored to see whether the iCBT program significantly contributes to their reduction .Automatic Thoughts Questionnaire (ATQ) [Attitude and Belief Scale-II (ABS-II) [Participants’ negative thoughts and related cognitive processes (most of them descriptions/inferences) were measured with the re (ATQ)  and irra(ABS-II) , 47. AltOnce registered on the treatment platform, participants received a unique study code. An independent person was asked to randomly assign the codes to one of two conditions  using a computer-generated randomization procedure with stratification by diagnostic status  as an additional criterion. The interviewers were blind to which group the participants had been randomized. However, because the difference between the iSOFIE and the control condition could not be masked, online therapists and participants were not blinded regarding the condition assignment.t-test and the Chi-square test. Because not all participants completed every weekly, post-test or follow-up measure, the missing data  were imputed with multiple imputation procedures [d for both within- and between-group comparisons, with the pooled standard deviation of the compared groups as denominator. Paired sample t-tests were used to test whether treatment gains were maintained between post-intervention and the six-month follow-up assessment. Finally, 95% confidence intervals (CI) were used to estimate the precision of the effect size for all measures. All analyses were performed in SPSS version 17.0 .At baseline, group differences in demographics were analyzed using the ocedures . No pattocedures . Effect Recovery rates based on patient’s self-reported social anxiety was estimated by the proportion of patients who scored below the clinical cut-off at the post-intervention assessment. Participants who did not meet the DSM-IV criteria for SAD at the post-intervention SCID interview  were considered responders. However, participants who did not meet the DSM-IV criteria for SAD but retained some residual symptoms at post-intervention were considered in partial remission. Clinically significant change was calculated on the main outcome measures using a conservative method suggested by Jacobson and Truax . The parF and χ2displaying a p between. 15 and. 81). Further details about the demographic characteristics are presented in The two conditions displayed similar demographic characteristics, and no statistically significant group differences were observed in terms of age, gender, educational level, professional and marital status, and previous psychotherapy attendance at baseline , and the number of completed modules is presented in the flow diagram . On average, participants completed 2.6 weekly worksheets. Overall, the iSOFIE participants completed 662 worksheets  were observed at post-intervention for all measures  of the participants in the iSOFIE group were considered symptom free  at the post-intervention SCID interview. Moreover, 14 additional participants (36.80%) significantly improved as they did not meet the DSM-IV criteria for SAD but maintained some residual social anxiety symptoms at the end of the treatment . Compared to the WLCG (n = 4), a significant proportion of the iSOFIE participants (n = 27) were considered responders as they obtained either total or partial remission  = 28.82; p<.01). Recovery rates show that 14 participants (36.80%) from the iSOFIE group and one participant (2.6%) from the WLCG score below the SPIN clinical cut-off  at post-intervention assessment. Most participants who scored below the SPIN clinical cut-off were also considered in total remission at the post-intervention SCID interview. Moreover, six-month later, in the absence of any intervention, the recovery rates ascended to 55.60% as 15 participants scored beyond the SPIN clinical cut-off.As concerning the treatment response, a total of 34.20% (χ2(1) = 10.53, p<.01.Additionally, the SPIN outcome measure showed that 44.7% (17/38) of the iSOFIE participants scores were clinically and significantly changed, and these results were different from the WLCG  = 13.30, p<.01).For the second treatment phase  44.7% (17/38) met the clinically significant change criterion for LSAS-SR and 60.5% (23/38) for SPIN.We further investigated the impact of the treatment using a clinical significant change criterion. The LSAS-SR outcome measure showed that47.4% (18/38) of the iSOFIE participants  reached this criterion, versus 13.2% (5/38) in the WLCG. This difference is statistically significant good and very good (28% and 72% respectively). The overall satisfaction with the program was also high, participants being satisfied or very satisfied with the intervention . Only a few responders declared to be neither satisfied nor dissatisfied with the program (14%), and none rated it as unsatisfactory. While the intervention was generally considered demanding or very demanding , all responders viewed the iSOFIE program as helpful or very helpful . On average participants spent 6.8 hours per week solving the various iSOFIE tasks  and most of them declared to have had an overall positive experience. Few complaints were made about the limited time available for each module (three participants), the perceived distant communication with her online therapists (one participant) and a remediable technical difficulty (one participant). Finally, participants were asked to indicate their level of agreement on a ten-point scale, where ten indicate the highest agreement. Participants rated the iSOFIE program approach as logical (9/10), and were willing to recommend it to a friend with similar problems (9/10).At the end of the program, participants who completed the treatment satisfaction questionnaire rated the quality of the iSOFIE modules as t-tests comparing pre- to post-intervention scores for the second treatment group revealed significant reductions for all measures (see last two rows of ES = 1.78 to 3.23) supporting once again the efficacy of this intervention (see n = 10) of the second treatment group participants scored below the SPIN clinical cut-off at post-intervention. No follow-up data were collected for this group.After the first group completed the intervention, participants from the wait-list control group were given the chance to receive the same intervention. Paired-sample tion see .RecoveryThis study was a randomized controlled trial aimed to investigate the efficacy of an internet based cognitive-behavioral intervention compared to a wait-list control group in the treatment of SAD. The Romanian iCBT intervention, named iSOFIE, was based on a Swedish internet-delivered CBT protocol which had been tested in previous studies , 39, 52.Overall, the results suggest that the iSOFIE program is effective not only in terms of treating SAD symptoms, but also in reducing depression and dysfunctional thinking . The intervention had a statistically significant effect on social anxiety symptoms, with large effect sizes for all the social anxiety symptoms measures. This supports previous findings presented in the literature , 53. FurIn terms of the secondary outcomes , significant changes favoring the iSOFIE group were observed, with moderate to large effect sizes. Since the changes in depression were obtained in the absence of direct focus of the treatment, these effects could show generalized effects of the intervention. Indeed, as depression and anxiety symptoms are often comorbid, they may have common cognitive mechanisms  that were addressed in the present study. Moreover, the treatment effects appear to be stable for both primary and secondary outcome measures, as most scores remained unchanged or decreased six months after the program ended. It is possible that, once participants did not meet the primary diagnostic criteria for SAD the secondary/associated depression symptoms were also reduced . Similar decreases in secondary outcome measures were found in previous studies. In termn = 204), a large between group ES at post-intervention was obtained on the LSAS-SR . Intere and. 99;). ConsidThis study presents a number of limitations. First, an active control condition matched for clients’ expectations, time, and therapist involvement was not included. Therefore the obtained results could be due to non-specific factors associated with any active treatment and the reported results should be interpreted with caution . Although we could not demonstrate that our iCBT program impacts participant’s life beyond the merely placebo effects, previous studies, 52, 54aInternet Social Phobia (iSOFIE). The overall results of the iSOFIE group were superior compared to the WLCG, satisfaction with the treatment being high. Comparable ES on both primary and secondary outcome measures were obtained with both the long and the short version of the SOFIE treatment manual. Overall, it seems that the iSOFIE intervention can be effectively used to overcome the problem of high prevalence versus low access to evidence-based treatment for SAD .These findings further support and replicate the efficacy of an iCBT program for SAD called To further advance the field of internet-delivered interventions future studies could include an active treatment condition, and investigate the external validity of such programs in psychiatric care. In this context, a tailored treatment might be needed in order to address potential co-morbid disorders . A progrS1 CONSORT Checklist(DOCX)Click here for additional data file.S1 Dataset(SAV)Click here for additional data file.S2 Dataset(SAV)Click here for additional data file.S1 Protocol(DOCX)Click here for additional data file.S2 Protocol(DOCX)Click here for additional data file.


Hepcidin reduces iron absorption by binding to the intestinal iron transporter ferroportin, thereby causing its degradation. Although short-term administration of testosterone or growth hormone (GH) has been reported to decrease circulating hepcidin levels, little is known about how hepcidin is influenced in human endocrine conditions associated with anemia.We used a sensitive and specific dual–monoclonal antibody sandwich immunoassay to measure hepcidin-25 in patients (a) during initiation of in vitro fertilization when endogenous estrogens were elevated vs. suppressed, (b) with GH deficiency before and after 12 months substitution treatment, (c) with hyperthyroidism before and after normalization, and (d) with hyperprolactinemia before and after six months of treatment with a dopamine agonist.In response to a marked stimulation of endogenous estrogen production, median hepcidin levels decreased from 4.85 to 1.43 ng/mL (p < 0.01). Hyperthyroidism, hyperprolactinemia, or GH substitution to GH-deficient patients did not influence serum hepcidin-25 levels.In humans, gonadotropin-stimulated endogenous estrogen markedly decreases circulating hepcidin-25 levels. No clear and stable correlation between iron biomarkers and hepcidin-25 was seen before or after treatment of hyperthyroidism, hyperprolactinemia or growth hormone deficiency. Hepcidin-25 plays a key role in the regulation of iron metabolism in humans by controlling the absorption of iron from the intestine . It is mHepcidin-25 was originally described as an antimicrobial protein belonging to the defensin group , 7. Howe.The endocrine system plays a major role in the regulation of erythropoiesis and this may in part be through effects on Hepcidin-25. For example, we have previously shown that 3 weeks of GH treatment in healthy subjects decreased hepcidin-25 concentrations by approximately 65%, presumably by stimulating erythropoiesis . In addiHAMP), and mice with disruption of the hepcidin gene expression do not display a polycythemic phenotype . Then, 100 μL hepcidin-25 calibrators (Peptides Insitute)  were added to the wells to generate a standard calibration curve. Serum samples were diluted with 20 parts assay buffer and added to their respective wells, and the ELISA plate was incubated for 1 h at room temperature. After aspiration, wells were washed 3 times with TBST, and 100 μL of a solution of conjugate antibody  diluted with 1000 parts TBST was added to each well for a 1-h incubation at room temperature. After aspiration, wells were washed 3 times with TBST, and the plate was developed with an MSD reader, which recorded ruthenium electrochemiluminescence.Hepcidin-25 concentrations were measured with a high-sensitivity sandwich ELISA specific for hepcidin-25, as described previously . In brieCRP concentrations were evaluated by immunoturbidimetry; this was done on a DxC/LX analyzer . Serum ferritin was determined on Beckman Coulter DXI by chemiluminescent immunoassay. Serum iron and TIBC were measured by spectrometry on a Beckman Coulter DXC 800 (Beckman Coulter Unicel DXC systems). All samples were analyzed according to the manufacturer’s advice at the Department of clinical chemistry, Karolinska University Hospital.Serum levels of free thyroxine (fT4) were measured using a modular analytics P170/P800 from Roche Diagnostics GmbH . Serum prolactin was measured using a commercial chemiluminescence immunoassay . Serum was not screened for macroprolactin due to the high specificity of the prolactin method. Free estradiol was measured using routine clinical assays  as described previously . Total sWilcoxon signed-rank test. Correlations were tested by calculation of the Spearman rank correlation coefficient. Data are expressed as median, (lower and upper quartile). Significance level was set at p < 0.05. Statistical analyses were performed using Statistica, Statsoft version 10.0 .Statistical analyses of hepcidin-25, CRP, ferritin, transferrin, TIBC and hormone concentrations were calculated using Altogether 31 healthy females were studied during suppression and stimulation of endogenous estrogen levels as part of pretreatment for IVF . There wPatients with GH deficiency all had markedly reduced IGF-I levels that were normalized following treatment with GH for one year . DespiteHyperthyroid patients had elevated fT4 and suppressed TSH levels (TSH data not shown) that were normalized following treatment . There wTreatment of hyperthyroidism decreased levels of ferritin significantly, and increased transferrin levels, see Patients with prolactinoma were studied before and after 6 months of therapy with dopamine agonist therapy, resulting in normalized prolactin levels . Also inCirculating hepcidin-25 is a major physiological regulator of iron metabolism and haematopoesis. In order to explore the possible influence of a set of human hormones on hepcidin-25 levels, we studied several endocrine perturbations in well-characterized clinical conditions. Of particular relevance is the fact that we were able to compare situations of over- or under-function in the same individuals, thereby eliminating the influence of inter-individual variation in hepcidin-25 levels, which is known to be wide , 28. OurPremenopausal non-pregnant women with regular menstruation cycles are often deficient in iron ; the oppIn male mice castration significantly increased hepcidin levels, whereas hepcidin mRNA levels were unchanged in ovariectomized female mice . Thus, tin vitro in human liver cells and in vivo in mice. An estrogen antagonist  prevented the estradiol-induced decrease and, independent of estradiol treatment, caused an increase in hepcidin mRNA, indicating that the estrogen receptor may have a ligand-independent effect on hepcidin expression. Our in vivo data support such a mechanism of estrogenic action on hepcidin-25 which in part seems to be independent of the moderate iron deficiency seen in our group of premenopausal, non-pregnant women.It has previously been shown that premenopausal women also have a lower hepcidin-25 level at the basal state  which isThe small increase in the CRP level during initiation of IVF did not correlate with estrogens levels (data not shown), supporting previous studies , 38 concWe have previously shown that daily administration of supraphysiological doses of GH to healthy volunteers for 3 weeks caused a sustained and significant suppression of hepcidin-25 concentrations . In thisIn the present work, ferritin levels were increased, and transferrin levels decreased, in the hyperthyroid state, without significant changes in hepcidin-25 or iron. This indicates that thyroid hormone does not primarily influence the hepatic production of hepcidin-25. We also explored whether there was any major influence of excess prolactin on hepcidin-25, but were not able to find any substantial change when this was normalized.per se [In conclusion, while testosterone per se , 11 has


Treponema pallidum PCR (Tp-PCR) has been noted as a valid method for diagnosing syphilis. We compared Tp-PCR to a combination of darkfield microscopy (DFM), the reference method, and serologic testing in a cohort of 273 patients from France and Switzerland and found the diagnostic accuracy of Tp-PCR was higher than that for DFM.  Treponema pallidum, has increased steadily worldwide since the early 2000s, especially in at-risk populations  to be a valid diagnostic method along with darkfield microscopy (DFM) ; most  were men. Mean delay from ulcer appearance to date of first medical visit was 20.4 days . Most patients were men who have sex with men (n = 185 [71.4%]). Ulcer localization was genital , anorectal , or oral . HIV status was known for 226 patients (82.8%); 53 were HIV positive, and 36 were receiving antiretroviral drug therapy. Nine patients received an initial HIV diagnosis at the time of the diagnostic work-up for syphilis.Tp-PCR specimens were positive; 13 of these were from patients who had negative DFM results. The proportion of negative DFM/positive Tp-PCR results was significantly higher for the 2 centers where DFM was performed only occasionally (6/15 [40.0%]) than for centers who performed DFM more often . The diagnostic performance of Tp-PCR against DFM was high (DFM results were assessed for 170 patients (62.3%); 32 had positive results (18.8%). Results for 43 was high , and agrTp-PCR were less sensitive and had a lower negative predictive value when serologic tests results were used as reference than when DFM results were used as reference (Tp-PCR provided higher specificity and positive predictive value when compared with this definition than when compared to either DFM or serologic test results alone (Tp-PCR and the enhanced definition (Specimens from 255 patients underwent serologic testing; 88 patients had positive results, and 16 patients had undetermined results. Results for eference . Under tts alone . When DFfinition , DFM senTp-PCR has a high degree of accuracy for the definitive diagnosis of primary syphilis from lesion exudate or tissue. As expected, the clinical value of this test appeared sensitive to the choice of reference test but was hampered by misclassification errors from DFM. By definition, any discrepancy between Tp-PCR and DFM results has been considered primarily an error in Tp-PCR. However, this assumption may not always be accurate. Our results demonstrate that Tp-PCR appeared higher, indicating that Tp-PCR has a high clinical usefulness either for confirming or for ruling out a suspicion of syphilis.The reliability of DFM in our study was strongly associated with routine performance. We classified cases with negative DFM results, positive serologic results, and a clinical picture evocative of syphilis as false negatives of the DFM. When we used this definition as a reference, the diagnostic performance of Tp-PCR in a unique laboratory. The study sample was also representative of patients who may benefit from Tp-PCR in the future. The main limitation was the lack of a standard protocol for serologic testing, which could have affected the validity of some analyses. However, we attempted to minimize intercenter variability by using a blind assessment of all serologic assays by 2 experts.The strengths of our study are its prospective and multicenter design and the performance of Tp-PCR provides better sensitivity/specificity than DFM when compared with clinical suspicion of syphilis (Tp-PCR compared with DFM (Our results concur with those of Grange et al., who reported that Currently, DFM is less often used in routine testing than it has been in the past (Tp-PCR as the reference diagnostic test for early-phase syphilis may be reasonable (Tp-PCR. First, Tp-PCR was more accurate than DFM when assessed against the enhanced definition in our study. Second, high-quality readings of DFM are difficult to obtain (Tp-PCR test relies less on human expertise than DFM, which may make Tp-PCR results more reproducible and testing less costly if it is performed on a routine basis.In summary, our results confirm that using Detailed methods and results for study of patients who had a genital, anal, or oral ulcer suggestive of syphilis after at-risk sexual intercourse during September 2011–September 2013 in 5 centers in Switzerland and France.