Label-Free Identification of Exosomes using Raman Spectroscopy and Machine Learning.

Exosomes, nano-sized extracellular vesicles (EVs) secreted from cells, carry various cargo molecules reflecting their cells of origin. As EV content, structure, and size are highly heterogeneous, their classification via cargo molecules by determining their origin is challenging. Here, a method is presented combining surface-enhanced Raman spectroscopy (SERS) with machine learning algorithms to employ the classification of EVs derived from five different cell lines to reveal their cellular origins. Using an artificial neural network algorithm, it is shown that the label-free Raman spectroscopy method's prediction ratio correlates with the ratio of HT-1080 exosomes in the mixture. This machine learning-assisted SERS method enables a new direction through label-free investigation of EV preparations by differentiating cancer cell-derived exosomes from those of healthy. This approach will potentially open up new avenues of research for early detection and monitoring of various diseases, including cancer.

The Microwave Temperature and Humidity Profiler: Description and Preliminary Results.

This manuscript presents the Microwave Temperature and Humidity Profiler (MTHP), a dual-band spectroradiometer designed for measuring multi-incidence angle temperature and humidity atmospheric profiles from an aircraft platform. The MTHP bands are at 60 GHz for measuring the oxygen complex lines, therefore at this band, MTHP has a hyperspectral radiometer able to provide 2048 channels over an 8 GHz bandwidth, and 183 GHz for measuring water vapor, which only uses four channels since this absorption band's spectral richness is simpler. The MTHP builds upon the Microwave Temperature Profiler (MTP) with the inclusion of the hyperspectral radiometer. The instrument's design, components, and calibration methods are discussed in detail, with a focus on the three-point calibration scheme involving internal calibration loads and static air temperature readings. Preliminary results from the Technological Innovation into Iodine and GV aircraft Environmental Research (TI3GER) campaign are presented, showcasing the instrument's performance during flights across diverse geographical regions. The manuscript presents successful antenna temperature measurements at 60 GHz and 183 GHz. The hyperspectral measurements are compared with a simulated antenna temperature using the Atmospheric Radiative Transfer Simulator (ARTS) showing an agreement better than R2 > 0.88 for three of the flights analyzed. Additionally, the manuscript draws attention to potential Radio Frequency Interference (RFI) effects observed during a specific flight, underscoring the instrument's sensitivity to external interference. This is the first-ever airborne demonstration of a broadband and hyperspectral multi-incidence angle 60 GHz measurement. Future work on the MTHP could result in an improved spatial resolution of the atmospheric temperature vertical profile and, hence, help in estimating the Planetary Boundary Layer (PBL) with better accuracy. The MTHP and its hyperspectral multi-incidence angle at 60 GHz have the potential to be a valuable tool for investigating the PBL's role in atmospheric dynamics, offering insights into its impact on Earth's energy, water, and carbon cycles.

Reliability and validity of the Turkish version of the Vocal Fatigue Index

Background/aim: The aim of this study is to evaluate the reliability and validity of the Turkish version of the Vocal Fatigue Index (VFI). Materials and methods: The study was conducted with 535 participants including 285 patients with voice disorders and 250 healthy controls. Internal consistency and test-retest reliability were calculated for the reliability analysis. The mean VFI factor scores of both groups were compared. The strength and direction of the relation between VFI and Voice-Related Quality of Life (V-RQOL) measure was evaluated for the validity analysis. Results: The Cronbach's alpha coefficient of VFI factor scores was found to be 0.920 for tiredness and avoidance of voice use, 0.879 for physical discomfort with voice use, and 0.882 for improvement of symptoms with voice rest. The test-retest reliability revealed 0.877 for the tiredness and avoidance of voice use, 0.913 for the physical discomfort with voice use, and 0.820 for the improvement of symptoms with voice rest. When compared with healthy individuals, VFI factor scores were statistically significant higher in patients with voice disorders. The V-RQOL scores decreased significantly as the VFI scores increased. Conclusion: The Turkish version of the VFI is a good reliable and valid instrument for evaluating vocal fatigue symptoms in the Turkish-speaking community.

[Investigation of congenital CMV infection with the presence of CMV DNA in saliva samples of new born babies]. / Yenidogan bebeklerin tükürük örneginde CMV DNA varligi ile konjenital CMV enfeksiyonunun arastirilmasi.

Cytomegalovirus (CMV), is the most common cause among congenital infections and is the most seen etiology in long-term sensorineural hearing loss (SNHL) and neurological impairment. Congenital CMV infection (CCMV) was reported in 0.15-2.2% of live-borne neonates in studies from different countries. A significant proportion of infected infants are asymptomatic after birth and might only be detected by routine screening methods during the new born period. The aim of this study was to screen the saliva of live-born neonates with areal-time PCR based method for the detection of CCMV in our hospital. Saliva samples collected in half an hour after birth by dry dacron swabs and were evaluated for CMV DNA (Rt-PCR, Abbott Molecular USA) from 1000 babies born in Ege University Faculty of Medicine Hospital Obstetrics Clinic between October 2015-October 2017. For the confirmation of CCMV, saliva positive newborns were evaluated with the same method for CMV DNA from their urine or blood within 21 days. All newborns were screened for sensorineural hearing tests. Subjects were 497 girls (49.7%) and 503 boys (50.3%), with a mean weight of 3116.8 g and mean of 37.61 birth week. CMV DNA was positive in the saliva of 16 newborns (1.6%). Fourteen newborns were weakly positive for CMV DNA in their saliva and were not confirmed for CCMV infection. Congenital CMV was confirmed in only two (0.2%) with the CMV DNA results in urine and/or blood samples. One of the two newborns with CCMV was symptomatic and had a neurosensorial hearing loss. The other one was asymptomatic. Saliva samples, taken immediately after birth with a noninvasive and easy method for the detection of CMV DNA is very important for diagnosis of CCMV. Positive samples should be confirmed with CMV DNA in urine or blood samples of these newborns. In this study, detection of positivity in saliva samples that were confirmed with other samples of our newborn population for CCMV was 0.2%. The specific diagnosis for CCMV in newborns with a noninvasive and easy collecting sample is important to avoid sequelae and for public health concerns.

The Effect of Mandibulectomy and Mandibulotomy on Cochlear Function.

Mandibulectomy and mandibulotomy procedures are performed with the oscillating saw and the acoustic energy generated during the osteotomies is transferred to the cochlea via temporomandibular joint and air conduction. The aim of this study was to assess the effect of mandibulectomy and mandibulotomy on cochlear function. This study was carried out on 15 patients who underwent at an otolaryngology department of a tertiary medical center between January 2013 and August 2015. The study was designed as a prospective study. All of the data were prospectively collected. These included demographic data, date of the surgery, type of surgery, preoperative pure-tone audiometry, preoperative and postoperative distortion product otoacoustic emission (DPOAE) measurements. Of the DPOAE measurements, there was a statistically significant difference between the signal-to-noise ratio measurement on the right ear measurement at 4 kHz (P <0.05). Additionally, there was a statistically significant correlation between the signal-to-noise ratio measurements and the side of the osteotomy (P <0.05). In conclusion, it is found that mandibulectomy and mandibulotomy procedure has a negative effect on cochlear function in the early period. The DPOAEs were diminished on the osteotomy site in day one postoperatively. Results with a longer follow-up time can yield more information on the prognosis of the cochlear damage.

The role of C-reactive protein/albumin ratio and prognostic nutritional index in the diagnosis of complicated acute appendicitis.

Objectives: C-reactive protein (CRP) levels increase and albumin levels decrease in patients with inflammation. CRP/albumin ratio (CAR) is a new inflammation-associated prognostic indicator. The prognostic nutritional index (PNI) was described as a simple and neutral indicator of adverse outcomes not only in chronic diseases but also in acute conditions. The aim of this study was to investigate the clinical significance of the CAR and PNI value in differentiating complicated acute appendicitis (AA). Material and Methods: We retrospectively examined the medical records of 187 patients with AA. Patients were divided into two groups according to pathological results [non-complicated (n= 161) and complicated (n= 26)]. Demographic, clinical, laboratory, and pathological data were examined and compared between the groups. Logistic regression analyses were performed to determine the independent predictors for complicated AA. Results: Median age of the study group was 32 (23-41) years, and most of the patients were males (n= 101, 54%). Patients in the complicated AA group were significantly older compared to the patients in the non-complicated AA group [38 (32-49.5) years vs. 30 (22-41) years, p= 0.002]. The complicated AA group had significantly higher CAR level compared to the non-complicated AA group (p= 0.001). The length of hospital stay was significantly longer in the complicated AA group compared to the non-complicated AA group [2.5 (2-4.25) days vs. 1 (1-2) days, p <0.001]. Other variables (including PNI) did not significantly differ between the groups. In univariate logistic regression analysis, only age was found to be a significant variable (OR= 1.045, 95% CI= 1.016-10.74, p= 0.002), but in multiple variate logistic regression analysis, no variable was found to be significant in predicting complicated AA. Conclusion: We concluded that CAR and PNI value are not independent predictors of complicated AA.

[A presentation of a practical algorithm which can be used in the management of vocal cord nodules, polyps and intraepithelial neoplasias]. / Vokal kord nodül, polip ve intraepitelyal neoplazilerinin yönetiminde kullanilabilecek pratik bir algoritma sunumu.

OBJECTIVES: This study aims to offer an accurate diagnosis for vocal cord nodules, polyps, and intraepithelial neoplasias through videolaryngostroboscopy (VLS) and multi-dimensional voice analysis program (MDVP) and to decide management modalities. PATIENTS AND METHODS: A total of 397 patients (214 males, 183 females; mean age 46.2 years; range 17 to 71 years) with 218 nodules, 101 polyps and 78 vocal cord intraepithelial neoplasias who were admitted for treatment and follow-up in phoniatry unit of our clinic in the past five years were retrospectively analyzed in terms of VLS findings, the percent of jitter, percent of shimmer, fundamentally frequency, noise-harmonic ratio values, GRBAS [Grade of dysphonia (G), roughness (R), breathiness (B), asthenicity (A) and strain (S)] scores, and voice handicap index (VHI) obtained by MDVP. RESULTS: We obtained better voice parameters with improved GRBAS and SHE scores in patients with vocal cord nodules who underwent voice training and those with polyps who were operated, while these scores worsened following diagnostic phonosurgery in the intraepithelial group. CONCLUSION: In the initial examination, patients who are prediagnosed vocal cord nodules should have voice therapy as the first-line treatment modality and checked for the response to treatment. If laryngeal cancer is suspected, surgery should be planned according to the biopsy result and biopsy should be performed, if polyp is present.

Automated Image Analysis for Characterization of Circulating Tumor Cells and Clusters Sorted by Magnetic Levitation.

Magnetic levitation-based sorting technologies have revolutionized the detection and isolation of rare cells, including circulating tumor cells (CTCs) and circulating tumor cell clusters (CTCCs). Manual counting and quantification of these cells are prone to time-consuming processes, human error, and inter-observer variability, particularly challenging when heterogeneous cell types in 3D clusters are present. To overcome these challenges, we developed "Fastcount," an in-house MATLAB-based algorithm for precise, automated quantification and phenotypic characterization of CTCs and CTCCs, in both 2D and 3D. Fastcount is 120 times faster than manual counting and produces reliable results with a ±7.3% deviation compared to a trained laboratory technician. By analyzing 400 GB of fluorescence imaging data, we showed that Fastcount outperforms manual counting and commercial software when cells are aggregated in 3D or staining artifacts are present, delivering more accurate results. We further employed Fastcount for automated analysis of 3D image stacks obtained from CTCCs isolated from colorectal adenocarcinoma and renal cell carcinoma blood samples. Interestingly, we observed a highly heterogeneous spatial cellular composition within CTCCs, even among clusters from the same patient. Overall, Fastcount can be employed for various applications with lab-chip devices, such as CTC detection, CTCC analysis in 3D and cell detection in biosensors.

Evaluation of Vestibulo-Ocular Reflex (VOR) in Tinnitus Patients with Normal Hearing.

OBJECTIVES: This study aimed to evaluate the correlation between tinnitus parameters (duration, severity, reaction, handicap levels) and vestibulo-ocular reflex (VOR) gain values in patients with tinnitus with normal hearing without vertigo or any other complaints and to compare the VOR gains with a healthy group. MATERIALS AND METHODS: The study group consisted of 30 individuals aged between 18 and 65 years who suffered from tinnitus but not from hearing loss and vertigo. The control group also consisted of 30 individuals who were categorized as healthy adults. The tinnitus handicap inventory, tinnitus reaction questionnaire, and tinnitus handicap questionnaire were applied to each individual in the tinnitus group, and the video head impulse test (VHIT) was conducted in 2 groups. RESULTS: Statistically, a significant difference was found between the 2 groups in terms of VOR gain values in horizontal and vertical semicircular canal planes (p<0.05). However, there was no statistically significant correlation between tinnitus parameters, age, and VOR gain values in the study group (p>0.05). CONCLUSION: Considering the lower VOR gain values of the study group than the control group, these patients may need to be followed up for vestibular dysfunction associated with tinnitus, which can be a symptom of peripheral vestibular disorder. Moreover, this study will contribute to the literature because we determined a high-frequency component of VOR by VHIT, which was used to evaluate the relationship between tinnitus parameters and peripheral vestibular function.

BIO-INSPIRED MAGNETIC BEADS FOR ISOLATION OF SPERM FROM HETEROGENOUS SAMPLES IN FORENSIC APPLICATIONS.

Rapid and efficient processing of sexual assault evidence will accelerate forensic investigation and decrease casework backlogs. The standardized protocols currently used in forensic laboratories require the continued innovation to handle the increasing number and complexity of samples being submitted to forensic labs. Here, we present a new technique leveraging the integration of a bio-inspired oligosaccharide (i.e., Sialyl-LewisX) with magnetic beads that provides a rapid, inexpensive, and easy-to-use strategy that can potentially be adapted with current differential extraction practice in forensics labs. This platform (i) selectively captures sperm; (ii) is sensitive within the forensic cut-off; (iii) provides a cost effective solution that can be automated with existing laboratory platforms; and (iv) handles small volumes of sample (∼200 µL). This strategy can rapidly isolate sperm within 25 minutes of total processing that will prepare the extracted sample for downstream forensic analysis and ultimately help accelerate forensic investigation and reduce casework backlogs.

Soft Ring-Shaped Cellu-Robots with Simultaneous Locomotion in Batches.

Untethered mini-robots can move single cells or aggregates to build complex constructs in confined spaces and may enable various biomedical applications such as regenerative repair in medicine and biosensing in bioengineering. However, a significant challenge is the ability to control multiple microrobots simultaneously in the same space to operate toward a common goal in a distributed operation. A locomotion strategy that can simultaneously guide the formation and operation of multiple robots in response to a common acoustic stimulus is developed. The scaffold-free cellu-robots comprise only highly packed cells and eliminate the influence of supportive materials, making them less cumbersome during locomotion. The ring shape of the cellu-robot contributes to anisotropic cellular interactions which induce radial cellular orientation. Under a single stimulus, several cellu-robots form predetermined complex structures such as bracelet-like ring-chains which transform into a single new living entity through cell-cell interactions, migration or cellular extensions between cellu-robots.

A confirmatory test for sperm in sexual assault samples using a microfluidic-integrated cell phone imaging system.

Rapid and efficient processing of sexual assault evidence to accelerate forensic investigation and decrease casework backlogs is urgently needed. Therefore, the standardized protocols currently used in forensic laboratories can benefit from continued innovation to handle the increasing number and complexity of samples being submitted to forensic labs. To our knowledge, there is currently no available rapid and portable forensic screening technology based on a confirmatory test for sperm identification in a sexual assault kit. Here, we present a novel forensic sample screening tool, i.e., a microchip integrated with a portable cell phone imaging platform that records and processes images for further investigation and storage. The platform (i) precisely and rapidly screens swab samples (<15 min after sample preparation on-chip); (ii) selectively captures sperm from mock sexual assault samples using a novel and previously published SLeX-based surface chemistry treatment (iii) separates non-sperm contents (epithelial cells and debris in this case) out of the channel by flow prior to imaging; (iv) captures cell phone images on a portable cellphone-integrated imaging platform, (v) quantitatively differentiates sperm cells from epithelial cells, using a morphology detection code that leverages Laplacian of Gaussian and Hough gradient transform methods; (vi) is sensitive within a forensic cut-off (>95% accuracy) compared to the manual counts; (vii) provides a cost-effective and timely solution to a problem which in the past has taken a great deal of time; and (viii) handles small volumes of sample (20 µL). This integration of the cellphone imaging platform and cell recognition algorithms with disposable microchips can be a new direction toward a direct visual test to screen and differentiate sperm from epithelial cell types in forensic samples for a crime laboratory scenario. With further development, this integrated platform could assist a sexual assault nurse examiner (SANE) in a hospital or sexual assault treatment center facility to flag sperm-containing samples prior to further downstream testing.

Levitating Cells to Sort the Fit and the Fat.

Density is a core material property and varies between different cell types, mainly based on differences in their lipid content. Sorting based on density enables various biomedical applications such as multi-omics in precision medicine and regenerative repair in medicine. However, a significant challenge is sorting cells of the same type based on density differences. Here, a new method for real-time monitoring and sorting of single cells based on their inherent levitation profiles driven by their lipid content is reported. As a model system, human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) from a patient with neutral lipid storage disease (NLSD) due to loss of function of adipose triglyceride lipase (ATGL) resulting in abnormal lipid storage in cardiac muscle are used. This levitation-based strategy detects subpopulations within ATGL-deficient hiPSC-CMs with heterogenous lipid content, equilibrating at different levitation heights due to small density differences. In addition, sorting of these differentially levitating subpopulations are monitored in real time. Using this approach, sorted healthy and diseased hiPSC-CMs maintain viability and function. Pixel-tracking technologies show differences in contraction between NLSD and healthy hiPSC-CMs. Overall, this is a unique approach to separate diseased cell populations based on their intracellular lipid content that cannot be achieved using traditional flow cytometry techniques.

Total Microfluidic chip for Multiplexed diagnostics (ToMMx).

Microfluidic technologies offer new platforms for biosensing in various clinical and point-of-care (POC) applications. Currently, at the clinical settings, the gold standard diagnostic platforms for multiplexed sensing are multi-step, time consuming, requiring expensive and bulky instruments with a constant need of electricity which makes them unsuitable for resource-limited or POC settings. These technologies are often limited by logistics, costly assays and regular maintenance. Although there have been several attempts to miniaturize these diagnostic platforms, they stand short of batch fabrication and they are dependent on complementary components such as syringe pumps. Here, we demonstrated the development and clinical testing of a disposable, multiplexed sensing device (ToMMx), which is a portable, high-throughput and user-friendly microfluidic platform. It was built with inexpensive plastic materials and operated manually without requiring electrical power and extensive training. We validated this platform in a small cohort of 50 clinical samples from patients with cardiovascular diseases and healthy controls. The platform is rapid and gives quantifiable results with high sensitivity, as low as 5.29 pg/mL, from only a small sample volume (4 µL). ToMMx platform was compared side-by-side with commercial ELISA kits where the total assay time is reduced 15-fold, from 5 h to 20 min. This technology platform is broadly applicable to various diseases with well-known biomarkers in diagnostics and monitoring, especially with potential future impact at the POC settings.

Evaluation of the Correlation Between Turkish Voice Handicap Index-10 and Turkish Voice-Related Quality of Life Scale.

OBJECTIVE: Aim of this sudy was to evaluate the correlation between Turkish Voice Handicap Index-10 (TVHI-10) and Turkish Voice-Related Quality of Life Questionnaries (TV-RQOL) on patients with dysphonia. METHODS: Two different groups were formed with patients with dysphonia and healthy individuals between February and July 2016. After medical history and otorhinolaringologic and phoniatric examinations all the subjects completed TVHI-10 and TV-RQOL questionnaries. The correlation between scores of parameters of questionnaries were evaluated. RESULTS: Data of 104 patients (59 women, 45 men) with a mean age of 46±15 years in dysphonia group and 75 individuals (38 women, 37 men) with a mean age of 45±13.8 years in healthy group were evaluated. TVHI-10 and TV-RQOL parameters' scores were significantly higher in dysphonia group than healthy group. There were positive and significant correlations between scores of TVHI-10 and TV-RQOL parameters of all 179 individuals. There was significant positive correlation between total scores of TVHI-10 and TV-RQOL values of all individuals (r=0.949, p<0.001). CONCLUSION: There is positive significant correlation between validated Turkish versions of VHI-10 and V-RQOL questionnaries as a self-assessment measurement tools. The results of studies which use TV-RQOL can be compared with the results of the studies using TVHI-10.

[Validation and reliability of Turkish Singing Voice Handicap index]. / Özgün San Sesi Handikap Endeksi ölçeginin Türkçe versiyonunun geçerlik ve güvenirligi.

OBJECTIVES: This study aims to constitute a valid and reliable Turkish version of the original Singing Voice Handicap Index. PATIENTS AND METHODS: An authorized committee assessed the reliability and validity of the content, scope, and language of the original Singing Voice Handicap Index which underwent a back translation process. The Turkish version of the questionnaire was answered twice with a 7 to 10-day interval by two singing voice groups with or without singing voice problems. The reliability and validity analyses were performed based on these answers. RESULTS: Of a total of 123 individuals (64 females, 59 males; mean age 26.2±7.3 years), 81 were without a voice pathology and 42 were with a voice pathology. The total Cronbach's alpha coefficient was 0.917. The item-total correlations ranged between 0.51 and 0.89. The weighted kappa values of test-retest correlation values of the items were 0.82-0.91. The Cronbach's alpha values of two part of the questionnaire based on the split-half method were 0.89 and 0.84. The mean total scale scores were 21.8±18.5 and 53.6±28.9 in normal and pathology groups, respectively and there was a statistically significant difference in scores between these two groups (p=0.000). CONCLUSION: The Turkish version of the Singing Voice Handicap Index is a valid and reliable scale which can be used in the evaluation of voice problems of Turkish-speaking singing voice users.