An improved tetra-primer PCR approach for the detection of the FGFR3 G380R mutation responsible for achondroplasia

Achondroplasia is the most common form of dwarfism and has an incidence of approximately 1/7500. In more than 98% of cases, the disease is associated with a G to A or G to C substitution at nucleotide position 1138 (p.G380R) of the fibroblast growth factor receptor 3 (FGFR3) gene. We have developed a sensitive single tube tetra-primer PCR assay to detect both the c.1138G>A and c.1138G>C mutations and can successfully distinguish DNA samples that are homozygous and heterozygous for the c.1138G>A mutation. Titration studies showed that the assay could reliably detect one copy of the mutant allele in a mix of 100 wild-type alleles. The assay has been tested in 50 healthy controls, 3 known patients with achondroplasia, and 5 amniotic fluids suspected of having achondroplasia and for whom we had previously determined the genotypes for the c.1138G>A mutation by PCR-RFLP. We have observed complete concordance between methods. Our tetra-primer PCR assay is sensitive, low-cost, and easy to use method for FGFR3 p.G380R genotyping, which could be used even in "low-tech" laboratories.     

Case Report: Acute Abdomen Caused by Brucellar Hepatic Abscess

Brucellosis, a zoonosis that is common worldwide, is endemic in many countries, primarily those of the Mediterranean region (including Turkey). Human brucellosis is a systemic infection with a wide clinical spectrum. Although hepatic involvement is very common during the course of chronic brucellosis, hepatic abscess is a very rare complication of Brucella spp. infection. We present a case of hepatic abscess caused by Brucella melitensis, which resembled the clinical presentation of surgical acute abdomen.     

To what extent and why are COPD and Willis-Ekbom disease associated?

Aim

Willis-Ekbom disease (RLS/WED) is common in chronic obstructive pulmonary disease (COPD). Patients with RLS/WED have poorer quality of sleep and more fatigue and depressive symptoms. The prevalence of RLS/WED in patients with COPD has been reported to vary between 29.1 and 36.8 %. However, during exacerbation, the prevalence can increase up to 54 %. These rates are higher than those seen in general population. We have not enough knowledge regarding the association between RLS and COPD. In this study, we aimed to determine the frequency of RLS in patients with stable COPD without comorbid conditions. In addition, we also aimed to determine possible related causative factors.

Method

We included 80 COPD patients without comorbid conditions who presented to our outpatient clinic between April 2013 and September 2013 for RLS/WED evaluation. Three cases that have polyneuropathy and one case that refused undergoing electromyography (EMG) examination were excluded from the study. Demographic data, P-A chest X-rays, pulmonary function tests (PFT), biochemical parameters (including hemogram), and dyspnea scales were evaluated for each patient. In addition, the RLS/WED rating scale and Epworth Sleep Scale (ESS) were applied. Further, each patient diagnosed with RLS/WED underwent a detailed neurological examination (performed by a neurologist) and an EMG examination to rule out polyneuropathy.

Results

Out of 76 COPD cases included in our study, 26.3 % (n?=?20) were diagnosed with RLS/WED (mean age 60.4?±?7.5 years, 20 males). The cases with RLS/WED had significantly lower body mass index (BMI) than cases without RLS/WED (p?=?0.009). There were no significant differences between cases with and without RLS/WED with respect to PFT, dyspnea scales, and arterial blood gas values. However, ESS was significantly different (p?=?0.016). There were no significant differences in RLS/WED scores and mean hs-CRP levels between COPD stages (p?=?0.424; p?=?0.518, respectively), while ESS was significantly different (p?=?0.016). ESS was significantly higher in stage B COPD than in stages A and D (p?=?0.005, p?=?0.008, respectively). Based on our model, we found that exacerbations and iron binding capacity (UIBC) were predictive factors for RLS/WED (p?<?0.100)

Conclusion

RLS/WED is a common disease in cases with stable COPD. Despite our hypothesis suggesting that the prevalence of RLS/WED in COPD is related with systemic inflammation, we did not find a significant association between hs-CRP and COPD cases with RLS/WED. However, we did find that UIBC is a predictive factor for the development of RLS/WED. Nonetheless, further studies are needed to understand the relationships between UIBC, low BMI, and the development of RLS/WED in COPD.

     

Effects of posterior capsular opacification on the evaluation of retinal nerve fiber layer as measured by Stratus optical coherence tomography

ObjectiveTo evaluate the effects of posterior capsular opacification (PCO) and Nd:YAG laser capsulotomy on the evaluation of peripapillary retinal nerve fibre layer (RNFL) as measured by Stratus (Carl Zeiss Meditec, Dublin, CA) optical coherence tomography (OCT).DesignProspective interventional case series.ParticipantsWe studied 98 eyes of 89 patients experiencing PCO.MethodsWe performed a complete ophthalmologic examination and evaluation of RNFL as measured by Stratus OCT before and after Nd:YAG capsulotomy. The patients were classified on the basis of PCO score or signal strength (SS) obtained by prelaser OCT scans.ResultsThe preoperative and postoperative peripapillary RNFL thicknesses were 90.24 ± 15.9 μm and 98.27 ± 14.1 μm, respectively (p < 0.001). The mean preoperative SSs were 5.4 ± 1.6, and they improved to 9.5 ± 0.5 postoperatively (p < 0.001). Prelaser and postlaser RNFL thicknesses differed significantly in eyes with preoperative SSs less than 7 (p < 0.05). Also, in patients with preoperative PCO grades higher than 2, there were significant increases in RNFL thickness after laser capsulotomy (p < 0.05).ConclusionRNFL thickness measured by the Stratus OCT is affected by PCO. RFNL thickness may be underestimated in eyes with preoperative PCO grades higher than 2 and in eyes with preoperative SSs lower than 7.     

Can rituximab be used in the treatment of pemphigus vulgaris during the COVID‐19 pandemic?

Rituximab is a monoclonal antibody that targets CD20, a B‐lymphocyte antigen; that leads to a decline in the B‐cell counts for at least a year. The patients who have received rituximab treatment in the previous 5 years with the diagnosis of pemphigus group of diseases at Cerrahpa?a Medical Faculty were questioned for COVID‐19 infection. A total of 48 patients were included in this study; only one male patient had COVID‐19 infection which had a mild course. There is no significant difference in the total number of lymphocytes between patients who have received rituximab within the previous 5 years or last year. The number of lymphocytes is independent of the number of courses of rituximab treatment received. Therefore, we suggest that all pemphigus patients who have received rituximab treatment within the previous 5 years should be careful of the preventive measures against the COVID‐19 infection irrespective of the number of treatment courses or the number of years which has passed since the treatment. The disease course was mild in the only infected patient. Thus, rituximab may be used in the treatment of pemphigus vulgaris during the COVID‐19 pandemic if its use is necessary.     

E-health literacy of nursing students and investigation of factors affecting e-health literacy during COVID-19 pandemic process: A cross-sectional study

The lack of knowledge on health literacy affects all segments of society, particularly health workers.The objectives were to identify nursing students’ means of accessing information during the coronavirus disease 2019 pandemic, their level of health literacy, and the factors that affect it.This cross-sectional study was conducted in Turkey among 398 nursing students of Çukurova University and Van Yüzüncü Yil University between June 1 and June 30, 2020. As a data collection tool, E-Health Literacy Scale was used, with students’ characteristics and personal information form related to Internet use. These forms were converted to the online format. The survey link was sent to the students’ smartphones and/or e-mails to ask them to participate.Majority of participants were Van Yüzüncü Yil University nursing students (63.8%). E-SYO score average of all students was found to be 29.42 ± 4.39 (min = 14, max = 40); it was is found be at a good level. They used the Internet as the first source of information about coronavirus disease 2019 (65.1%).Among the participants, 65.8% stated that it was important to access the health resource on the Internet and 19.1% of the participants thought that it was very important. It was found that Internet use was being used for >3 times a day (72.9%). The age, class, gender, family type, income level, high school from which they graduated from, and their working status significantly were statistically affecting their health literacy (P < .05). The health literacy scale scores were significant and higher than those who did not know the concept of health literacy, and those who perceived Internet skills well and very well than those who perceived them poorly (P < .05).Nursing students were found to have good average health literacy averages. Improving the health literacy is important for making individuals healthier.     

High contrast,isotropic, and uniform 3D-imaging of centimeter-scale scattering samples using structured illumination light-sheet microscopy with axial sweeping

Light-sheet fluorescent microscopy (LSFM) has, in recent years, allowed for rapid 3D-imaging of cleared biomedical samples at larger and larger scale. However, even in cleared samples, multiple light scattering often degrades the imaging contrast and widens the optical sectioning. Accumulation of scattering intensifies these negative effects as light propagates inside the tissue, which accentuates the issues when imaging large samples. With axially swept light-sheet microscopy (ASLM), centimeter-scale samples can be scanned with a uniform micrometric optical sectioning. But to fully utilize these benefits for 3D-imaging in biomedical tissue samples, suppression of scattered light is needed. Here, we address this by merging ASLM with light-sheet based structured illumination into Structured Illumination Light-sheet Microscopy with Axial Sweeping (SILMAS). The SILMAS method thus enables high-contrast imaging, isotropic micrometric resolution and uniform optical sectioning in centimeter-scale scattering samples, creating isotropic 3D-volumes of e.g., whole mouse brains without the need for any computation-heavy post-processing. We demonstrate the effectiveness of the approach in agarose gel phantoms with fluorescent beads, and in an PFF injected alpha-synuclein transgenic mouse model tagged with a green fluorescent protein (SynGFP). SILMAS imaging is compared to standard ASLM imaging on the same samples and using the same optical setup, and is shown to increase contrast by as much as 370% and reduce widening of optical sectioning by 74%. With these results, we show that SILMAS improves upon the performance of current state-of-the-art light-sheet microscopes for large and imperfectly cleared tissue samples and is a valuable addition to the LSFM family.