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1.

Background

Understanding factors that influence public support for “nudging” policies, like pictorial cigarette pack warnings, may offer insight about how to increase such support. We sought to examine factors that influence smokers’ support for requiring pictorial warnings on cigarette packs.

Methods

In 2014 and 2015, we randomly assigned 2149 adult US smokers to receive either pictorial warnings or text-only warnings on their cigarette packs for 4 weeks. The outcome examined in the current study was support for a policy requiring pictorial warnings on cigarette packs in the US.

Results

Support for pictorial warnings was high at baseline (mean: 3.2 out of 4). Exposure to pictorial warnings increased policy support at week 4 (β = .05, p = .03). This effect was explained by increases in perceived message effectiveness (p < .001) and reported conversations about policy support (p < .001). Message reactance (i.e., an oppositional reaction to the warning) partially diminished the impact of pictorial warnings on policy support (p < .001).

Conclusions

Exposing people to a new policy through implementation could increase public support for that policy by increasing perceived effectiveness and by prompting conversations about the policy. Reactance may partially weaken the effect of policy exposure on public support.
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2.

?

Helicobacter bilis is a commensal bacterium causing chronic hepatitis and colitis in mice. In humans, enterohepatic Helicobacter spp. are associated with chronic hepatobiliary diseases.

Purpose

We aimed at understanding the microbial etiology in a patient with X-linked agammaglobulinemia presenting with suppurative cholangitis.

Methods

16S rDNA PCR directly performed on a liver biopsy retrieved DNA of H. bilis.

Results

Clinical outcome resulted in the normalization of clinical and biological parameters under antibiotic treatment by a combination of ceftriaxone, metronidazole, and doxycyclin followed by a 2-week treatment with moxifloxacin and a 2-month treatment with azithromycin.

Conclusion

In conclusion, these data suggest a specific clinical and microbiological approach in patients with humoral deficiency in order to detect H. bilis hepatobiliary diseases.
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3.

Background

Recently, two functional IL18 promoter variants, ?607C>A (rs1946518) and ?137G>C (rs187238), were associated with viral clearance in patients with hepatitis C. The present study focused on their relevance for treatment response.

Methods

Seven hundred fifty-seven chronically infected European patients and 791 controls were enrolled in the study. IL18 genotyping was performed by allele-specific PCR. Liver histology was available in 67.9%.

Results

Genotype and allele frequencies were equally distributed in patients and controls. No significant association with various disease characteristics was observed. However, when comparing patients with sustained virological response (SR) and non-SR, statistically significant associations were found for both variants (p?=?0.0416 and p?=?0.0274, respectively). In viral genotype 1, the ?607A allele was positively associated with treatment response (p?=?0.0190; OR 1.537; 95% CI, 1.072–2.205) and the ?137G allele with a higher rate of nonresponse (p?=?0.0302; OR 1.524; 95% CI, 1.040–2.233).

Conclusions

The association of IL18 variants with treatment response in genotype 1 hepatitis C patients implies a predictive and modifying role of these genetic variants.
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4.
5.

Introduction and objective

Helicobacter pylori (H. pylori) is a human gastric pathogen. Because of presence of H. pylori oral cavity, there is a possibility of H. pylori transmission by oral-oral route. In a crosssectional study, the presence of some virulence factors of H. pylori and also non-pylori Helicobacters in dental plaque samples of participants was investigated.

Methods

The samples were collected from at least two teeth surfaces. DNA of the samples was extracted using specific kit. The presence of dupA (jhp0917 and jhp0918) and babA2 genes and also Helicobacter genus and H. pylori species was investigated using polymerase chain reaction by specific primers.

Results

In total 44% (20/45) and 86.7% (39/45) of samples was positive for ureC and 16SrRNA genes respectively. The frequency of babA2, jhp0917 and jhp0918 genes in H. pylori isolates were 40, 20 and 65% respectively. It seems 19 samples were positive probably non-pylori Helicobacters.

Conclusion

In the present study we report for the first time the presence of non-pylori Helicobacter species and also high frequency of babA2 and dupA genotypes in dental plaque samples.
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6.

Background

To investigate the expression of chemokine ligand 2 (CCL2), chemokine ligand 18 (CCL18), and vascular endothelial growth factor (VEGF) in peripheral blood of patients with gastric cancer and their correlation with presence of malignancy and disease progression.

Methods

Sixty patients with pathological proved gastric cancer were prospectively included into study. The levels of CCL2, CCL18, and VEGF in peripheral blood were examined by enzyme-linked immunosorbentassay (ELISA). Peripheral blood from 20 healthy people was examined as control.

Results

The preoperative serum levels of CCL2, CCL18 and VEGF in gastric cancer patients were significantly higher than that of controls (P <0.001, P <0.001, and P <0.001, respectively). ROC curve analysis showed that with a cut-off value of ≥1272.8, the VEGF*CCL2 predicted the presence of gastric cancer with 83% sensitivity and 80% specificity. Preoperative serum CCL2 was significantly correlated to N stage (P =0.040); CCL18 associated with N stage (P =0.002), and TNM stage (P =0.002); VEGF correlated to T stage (P =0.000), N stage (P =0.015), and TNM stage (P =0.000).

Conclusion

Preoperative serum levels of CCL2 and VEGF could play a crucial role in predicting the presence and progression of gastric cancer.
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7.

Background

Postmenopausal women experience estrogen deficiency-related menopausal symptoms (e.g., hot flashes and mood swings) and a dramatic increase in the incidence of chronic diseases. Although estrogen-replacement therapy (ERT) can reduce mortality from cardiovascular disease and improve osteoporosis and menopausal symptoms, its side effects have limited recent use. This study investigated the estrogen-like activity of aqueous extract from Agrimonia pilosa Ledeb.

Methods

The estrogenic activity of A. pilosa was investigated by using several in vitro assays. The binding activity of A. pilosa on estrogen receptors was examined using a fluorescence polarization-based competitive binding assay. The proliferative activity of A. pilosa was also examined using MCF-7 cells. Furthermore, the effect of A. pilosa on the expression of 3 estrogen-dependent genes was assessed.

Results

Using liquid chromatography-mass spectrometry, the 3 major peaks of A. pilosa aqueous extract were identified as apigenin-hexose, luteolin-glucuronide, and apigenin-glucuronide. The aqueous extract induced the proliferation of estrogen receptor-positive MCF-7 cells (p?<?0.05). A. pilosa-stimulated proliferation was blocked on adding the estrogen antagonist ICI 182,780. Moreover, A. pilosa treatment increased the mRNA expression of the estrogen-responsive genes pS2 and PR (p?<?0.05).

Conclusions

These results suggest A. pilosa can be used to improve estrogen deficiency-related menopausal symptoms or to treat diseases in postmenopausal women.
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8.

Background

Mutations in the cyclin-dependent kinase-like 5 gene cause a clinical variant of Rett syndrome (CDKL5-RTT). A role for the acute-phase response (APR) is emerging in typical RTT caused by methyl-CpG-binding protein 2 gene mutations (MECP2-RTT). No information is, to date, available on the inflammatory protein response in CDKL5-RTT. We evaluated, for the first time, the APR protein response in CDKL5-RTT.

Methods

Protein patterns in albumin- and IgG-depleted plasma proteome from CDKL5-RTT patients were evaluated by two-dimensional gel electrophoresis/mass spectrometry. The resulting data were related to circulating cytokines and compared to healthy controls or MECP2-RTT patients. The effects of omega-3 polyunsaturated fatty acids (ω-3 PUFAs) were evaluated.

Results

CDKL5-RTT mutations resulted in a subclinical attenuated inflammation, specifically characterized by an overexpression of the complement component C3 and CD5 antigen-like, both strictly related to the inflammatory response. Cytokine dysregulation featuring a bulk increase of anti-inflammatory cytokines, predominantly IL-10, could explain the unchanged erythrocyte sedimentation rate and atypical features of inflammation in CDKL5-RTT. Omega-3 PUFAs were able to counterbalance the pro-inflammatory status.

Conclusion

For the first time, we revealed a subclinical smouldering inflammation pattern in CDKL5-RTT consisting in the coexistence of an atypical APR coupled with a dysregulated cytokine response.
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9.

Purpose

This study examined the developmental trajectories of self-esteem and body mass index (BMI) from adolescence to mid-adulthood and the way the association between self-esteem and BMI changed during a 26-year follow-up.

Methods

Participants of a Finnish cohort study in 1983 at 16 years (N?=?2194) were followed up at ages 22 (N?=?1656), 32 (N?=?1471), and 42 (N?=?1334) using postal questionnaires. Measures at each time point covered self-esteem and self-reported weight and height. Analyses were done using latent growth curve models (LGM) and difference scores.

Results

In LGM analyses among females both the initial levels (r?=??0.13) and slopes (r?=?-0.26) of the self-esteem and BMI trajectories correlated negatively. Among males, there were no significant correlations between self-esteem and BMI growth factors. The association between increasing BMI and decreasing self-esteem among females was strongest between ages 22 and 32 (r?=??0.16), while among males, increases in BMI and self-esteem correlated positively (r?=?0.11) during that period. Among females, cross-sectional correlations between self-esteem and BMI showed an increasing trend (p?<?0.001) from age 16 (r?=??0.07) to age 42 (r?=??0.17), whereas among males negative correlation (r?=??0.08) emerged only in mid-adulthood at age 42.

Conclusion

Among females, higher and increasing BMI is associated with lower and more slowly increasing self-esteem. This association is not restricted to adolescent years but persists and gets stronger in mid-adulthood. Among males, associations are weaker but indicate more age-related differences. The results highlight the need for interventions that tackle weight-related stigma and discrimination, especially among women with higher body weight and size.
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10.

Objective

The functional PTPN22 R620W polymorphism (rs2476601) is clearly associated with susceptibility to several autoimmune diseases (ADs). However, the PTPN22 R263Q polymorphism (rs33996649) has been scarcely explored in different ADs. Here we aimed to examine the associations of the PTPN22 R620W and R263Q polymorphisms with susceptibility to or protection against rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and Graves’ disease (GD) among Mexican patients.

Methods

We conducted a case–control study including 876 patients (405 with SLE, 388 with RA, and 83 with GD) and 336 healthy control individuals. PTPN22 genotypes were determined using the TaqMan 5′ allele discrimination assay.

Results

PTPN22 R620W was associated with GD susceptibility (OR 4.3, p = 0.004), but was not associated with SLE (OR 1.8, p = 0.19). We previously demonstrated that this polymorphism is associated with RA susceptibility (OR 4.17, p = 0.00036). Moreover, PTPN22 R263Q was associated with protection against SLE (OR 0.09, p = 004) and RA (OR 0.28, p = 0.045), but was not associated with GD.

Conclusions

Our data provide the first demonstration that PTPN22 R620W confers GD susceptibility among Latin-American patients. Moreover, this is the second report documenting the association of PTPN22 R263Q with protection against SLE and RA.
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11.

Purpose

Vaccination uptake remained low, although annual subsidies are provided to encourage 6–72-month-old Hong Kong children to be vaccinated against seasonal influenza. This study was aimed to investigate the psychosocial influences on parental decision-making regarding young children’s seasonal influenza vaccination.

Methods

One-thousand two-hundred twenty-six parents of eligible children were recruited using random digit dialing in August–October 2012 to assess baseline perceptions and re-contacted in March 2013 to record children’s vaccination uptake. Structural equation modeling (SEM) was performed to examine factors associated with parental decision about children’s vaccination based on the complete data of 1222 respondents.

Results

Of the 1226 respondents who completed the follow-up survey, 34.3 % reported that their child was vaccinated during the follow-up period. Child’s past influenza vaccination history (β?=?0.48), belief in vaccination safety (β?=?0.35), and social norms (β?=?0.25) were strongly associated with parental intention to vaccinate their child which directly predicted child vaccination uptake (β?=?0.57). Belief in vaccination safety (β?=?0.42) and social norms (β?=?0.36) were strongly associated with vaccination intention of parents whose children never received influenza vaccine.

Conclusion

Interventions that address concerns on vaccination safety and utilize social norms may be effective to initiate Chinese parents to vaccinate their children.
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12.

Objective

The aim of this study was to investigate the involvement of TLR adaptor molecules, such as TRIF, MyD88, and TBK1 in the induction of iNOS and nitric oxide (NO) production in Pam2CSK4 and Pam3CSK4-treated mouse macrophages.

Method

Mouse macrophage cell line (RAW264.7) was transfected with trif, myd88, and tbk1 siRNAs before stimulated with Pam2CSK4 and Pam3CSK4. The iNOS gene and protein expression were determined by RT-PCR and immunoblotting, respectively. The NO production was determined by Griess reaction assay.

Results

The results showed that the induction of iNOS expression and NO production by Pam2CSK4 and Pam3CSK4 were diminished in tbk1 and myd88-depleted mouse macrophages but not trif-depleted cells.

Conclusion

These results suggested that the TBK1 and MyD88 molecules were essential for the induction of iNOS expression and NO production by both Pam2CSK4 and Pam3CSK4 via TLR2 signaling.
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13.

Objective

This study aimed at investigating the in vitro activity of minocycline and doxycycline on human polymorphonuclear (h-PMN) cell function.

Methods

h-PMNs were isolated from whole venous blood of healthy subjects; PMN oxidative burst was measured by monitoring ROS-induced oxidation of luminol and transendothelial migration was studied by measuring PMN migration through a monolayer of human umbilical vein endothelial cells. Differences between multiple groups were determined by ANOVA followed by Tukey’s multiple comparison test; Student’s t test for unpaired data for two groups.

Results

Minocycline (1–300 µM) concentration dependently and significantly inhibited oxidative burst of h-PMNs stimulated with 100 nM fMLP. Ten micromolar concentrations, which are superimposable to C max following a standard oral dose of minocycline, promoted a 29.8 ± 4 % inhibition of respiratory burst (P < 0.001; n = 6). Doxycycline inhibited ROS production with a lesser extent and at higher concentrations. 10–100 µM minocycline impaired PMN transendothelial migration, with maximal effect at 100 µM (42.5 ± 7 %, inhibition, n = 5, P < 0.001).

Conclusions

These results added new insight into anti-inflammatory effects of minocycline exerted on innate immune h-PMN cell function.
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14.

Purpose

The purpose of this study was to investigate the impact of malignancy and chemotherapy on the clinical and microbiological characteristics of Clostridium difficile infections (CDI).

Methods

CDI patients with a history of malignancy within 5 years were defined as the cancer group. The characteristics of the patients were compared according to the presence of malignancy.

Results

Of 580 patients with CDI, 159 (27.4 %) belonged to the cancer group and 421 (72.6 %) to the non-cancer group. More of the patients in the cancer group than those in the non-cancer group had been hospitalized within the prior 2 months (P?<?0.001). Leukocytosis was more common in the non-cancer group (P?=?0.034), while infection by PCR ribotype 017 strains was more common in the cancer group, with marginal significance (P?=?0.07). Recurrence was more frequent in the cancer group (20.4 % vs. 9.5 %, P =0.005) and cancer was an independent risk factor for recurrence of CDI (OR?=?2.66, 95 % CI 1.34-5.29, P =0.005). Age also contributed to the recurrence of CDI (OR?=?1.03, 95 % CI 1.00-1.06, P =0.026).

Conclusions

Malignancy and age are independent risk factors for recurrence of CDI. Cancer patients require careful observation for recurrence after treatment of CDI.
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15.

Background

The focus on translational research in clinical trials has the potential to generate clinically relevant genetic data that could have importance to patients. This raises challenging questions about communicating relevant genetic research results to individual patients.

Methods

An exploratory pharmacogenetic analysis was conducted in the international ovarian cancer phase III trial, AGO-OVAR 16, which found that patients with clinically important germ-line BRCA1/2 mutations had improved progression-free survival prognosis. Mechanisms to communicate BRCA results were evaluated, because these findings may be beneficial to patients and their families.

Results

Communicating individual BRCA results was not anticipated during clinical trial design. Consequently, options were not available for patients to indicate their preference for receiving their individual results when they signed pharmacogenetic informed consent. Differences in local requirements, clinical practice, and opinion regarding the ethical aspects of how to convey genetic results to patients are all potential barriers to returning individual BRCA results to patients. Communicating the aggregate BRCA result from this study provided clinical investigators with a mechanism to disseminate the overall study finding to patients while taking individual circumstances, local guidelines and clinical practice into account.

Conclusion

This study illustrates the importance of increasing the clarity and scope of informed consent and the need for patient engagement to ensure clinical trial participants can indicate their preference regarding receipt of potentially important individual pharmacogenetic results.

Trial registration

This study was registered in the NCT Clinical Trial Registry under NCT00866697 on March 19, 2009, following approval from participating ethics committees (Additional file 1).
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16.

Background

Obesity is one of the main causes of morbidity and mortality worldwide. More than 120 genes have been shown to be associated with obesity related phenotypes. The aim of this study was to determine the effect of selected genetic polymorphisms in Uncoupling protein 1 (UCP1) and Niemann-Pick C1 (NPC1) genes in an obese population in Saudi Arabia.

Methods

The genotypes of rs1800592, rs10011540 and rs3811791 (UCP1 gene) and rs1805081 and rs1805082 (NPC1 gene) were determined in a total of 492 subjects using TaqMan chemistry by Real-time PCR. In addition, capillary sequencing assay was performed to identify two specific polymorphisms viz., rs45539933 (exon 2) and rs2270565 (exon 5) of UCP1 gene.

Results

A significant association of UCP1 polymorphisms rs1800592 [OR, 1.52 (1.10–2.08); p?=?0.009] was observed in the obese cohort after adjusting with age, sex and type 2 diabetes. Further BMI based stratification revealed that this association was inconsistent with both moderate and extreme obese cohort. A significant association of UCP1 polymorphisms rs3811791 was observed only in the moderate-obese cohort [OR?=?2.89 (1.33–6.25); p?=?0.007] but not in the extreme-obese cohort indicating an overlying genetic complexity between moderate-obesity and extreme-obesity. The risk allele frequencies, which were higher in moderate-obese cohort, had abnormal HDL, LDL and triglyceride levels.

Conclusion

The rs1800592 and rs3811791 of UCP1 gene are associated with obesity in general and in the moderate-obese group in particular. The associated UCP1 polymorphisms in the moderate-obese group may regulate the impaired energy metabolism which plays a significant role in the initial stages of obesity.
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17.

Background

We examined the therapeutic potential of a proprietary Croton palanostigma extract (Zangrado®) in the management of emesis and itch.

Methods

Emesis was induced in ferrets with morphine-6-glucuronide (0.05 mg/kg sc) in the presence of Zangrado (3 mg/kg, ip) and the cannabinoid receptor 1 antagonist, AM 251 (5 mg/kg, ip). Topical Zangrado (1%) was assessed for anti-pruretic actions in the 5-HT-induced scratching model in rats and evaluated in capsaicin-induced gastric hyperemia as measured by laser doppler flow. In the Apc Min mouse model of precancerous adenomatosis polyposis, mice received Zangrado (100 μg/ml in drinking water) from the age of 6 – 16 weeks for effects on polyp number. In RAW 264.7 cells Zangrado was examined for effects on lipopolysaccharide-induced nitrite production.

Results

Zangrado was a highly effective anti-emetic, reducing morphine-induced vomiting and retching by 77%. These benefits were not associated with sedation or hypothermia and were not reversed by cannabinoid receptor antagonism. Itch responses were blocked in both the morphine and 5-HT models. Zangrado did not exacerbate the Apc Min condition rather health was improved. Capsaicin-induced hyperemia was blocked by Zangrado, which also attenuated the production of nitric oxide by activated macrophages.

Conclusion

Zangrado is an effective anti-emetic and anti-itch therapy that is devoid of common side-effects, cannabinoid-independent and broadly suppresses sensory afferent nerve activation. This complementary medicine represents a promising new approach to the management of nausea, itch and irritable bowel syndrome.
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18.

Background

Physical examinations may reveal the instability of a glenohumeral joint but cannot diagnose the bony Bankart lesions. Soft tissue Bankart lesion cannot be visualized on traditional radiogram. Magnetic resonance images have high cost and availability issues. The purpose of the study was to access the diagnostic performance of the Computed Tomography (CT) in the assessment of patients with shoulder instability and to diagnose the Bankart and bony Bankart lesions.

Methods

A total of 145 patients with shoulder instability were included in the study. Patients were subjected to clinical examination tests, traditional radiography, and CT. Two orthopedic surgeons, two engineers (trained in musculoskeletal imaging), and two physiotherapists have analyzed the radiological images, CT scans, and the clinical examination tests respectively. The Chi-square test or one-way ANOVA/ Dunnett Multiple comparisons test was performed at 99% of confidence level.

Results

Sensitivity (0.972?±?0.18 vs. 1, p?=?0.11) and accuracy (0.942?±?0.17 vs. 1, p?<?0.0001, q?=?3.88) for the clinical examination tests combining the traditional radiological images were same to CT. However, the clinical examination tests combining the traditional radiological images had more inconclusive results (5 vs. 1), false-positive results (6 vs. 5), and false negative results (4 vs. 1) than CT. The area that detects the Bankart and bony Bankart lesions at least one time for CT was higher than that of the clinical examination tests combining the traditional radiological images.

Conclusion

CT should be considered for evaluation in patients with shoulder instability and suspected Bankart and bony Bankart lesions.

Trial registration

Researchregistry3990 dated 15 December 2014 (www.researchregistry.com).
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19.

Background

International Service Learning Trips (ISLT) provide health professional students the opportunity to provide healthcare, under the direction of trained faculty, to underserved populations in developing countries. Despite recent increases in international service learning trips, there is scant literature addressing concerns students have prior to attending such trips. This study focuses on identifying concerns before and after attending an ISLT and their impact on students.

Methods

A survey comprised of closed and open-ended questions was developed to elucidate student concerns prior to attending an ISLT and experiences which might influence concerns. A five-point Likert-scale (extremely concerned?=?1, minimally concerned?=?5) was used to rate apprehension and satisfaction. Paired t-test was used to compare pre- and post-trip concerns; Chi-Square test was used to compare groups.

Results

Thirty-five students (27 medical, 8 pharmacy) attended ISLTs in December 2013. All completed pre and post-trip surveys. Significant decreases were seen in concerns related to cultural barriers (4.14 vs 4.46, P?=?.047), disease/epidemics (3.34 vs 4.60, P?<?.001), natural disasters (3.94 vs 4.94, P?<?.001), terrorism (4.34 vs 4.94, P?<?.001), travel (3.86 vs 4.51, P?<?.001) monetary issues (3.80 vs 4.60, P?<?.001), hospitality (3.94 vs 4.74, P?=?.001) and food (3.83 vs 4.60, P?<?.001). Language and group dynamics remained concerns post-trip. On open-ended questions, students described benefits of attending an ISLT.

Conclusions

Students had multiple concerns prior to attending an ISLT. Most decreased upon return. Addressing concerns has the potential to decrease student apprehension. The results of this study highlight the benefits of providing ISLTs and supporting development of a curriculum incorporating trip-related concerns.
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20.

Background

The human leukemia cell line HL-60 is considered an alternative cell culture model to study neutrophil differentiation and migration. The aim of this study was to characterize the suitability of HL-60 cells differentiated to neutrophil-like cells (nHL-60) as substitute for blood-derived human neutrophils to investigate the interaction of neutrophils with Staphylococcus aureus.

Methods

For this purpose, antimicrobial activity, bacterial uptake, production of reactive oxygen species and the release of neutrophil extracellular traps (NETs) by nHL-60 cells were analyzed and compared to primary blood-derived neutrophils using Staphylococcus aureus as important human and animal pathogen.

Results

Overall, the antimicrobial activities of nHL-60 cells were distinctly lower compared to blood-derived neutrophils. Furthermore, production of reactive oxygen species as well as NET formation was clearly impaired in nHL-60 cells.

Conclusion

This study indicates that HL-60 cells are of limited usage as an alternative model to study antimicrobial functions of neutrophils against Staphylococcus aureus.
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