Subtyping on Live Lymphoma Cell Lines by Raman Spectroscopy

Raman spectroscopy is a well-defined spectroscopic technique sensitive to the molecular vibrations of materials, since it provides fingerprint-like information regarding the molecular structure of the analyzed samples. It has been extensively used for non-destructive and label-free cell characterization, particularly in the qualitative and quantitative estimation of amino acids, lipids, nucleic acids, and carbohydrates. Lymphoma cell classification is a crucial task for accurate and prompt lymphoma diagnosis, prognosis, and treatment. Currently, it is mostly based on limited information and requires costly and time-consuming approaches. In this work, we are proposing a fast characterization and differentiation methodology of lymphoma cell subtypes based on Raman spectroscopy. The study was performed in the temperature range of 15–37 °C to identify the best cell measurement conditions. The proposed methodology is fast, accurate, and requires minimal sample preparation, resulting in a potentially promising, non-invasive strategy for early and accurate cell lymphoma characterization.

Importance of Adequate qPCR Controls in Infection Control

Respiratory screening assays lacking Sample Adequacy Controls (SAC) may result in inadequate sample quality and thus false negative results. The non-adequate samples might represent a significant proportion of the total performed tests, thus resulting in sub-optimal infection control measures with implications that may be critical during pandemic times. The quantitative sample adequacy threshold can be established empirically, measuring the change in the frequency of positive results, as a function of the numerical value of “sample adequacy”. Establishing a quantitative threshold for SAC requires a big number/volume of tests to be analyzed in order to have a statistically valid result. Herein, we are offering for the first time clear clinical evidence that a subset of results, which did not pass minimal sample adequacy criteria, have a significantly lower frequency of positivity compared with the “adequate” samples. Flagging these results and/or re-sampling them is a mitigation strategy, which can dramatically improve infection control measures.

Molecular and MALDI-TOF MS identification of swallow bugs Cimex hirundinis (Heteroptera: Cimicidae) and endosymbionts in France

Background The Cimicidae are obligatory blood-feeding ectoparasites of medical and veterinary importance. We aim in the current study to assess the ability of MALDI-TOF MS to identify Cimex hirundinis swallow bugs collected in house martin nests. Methods Swallow bugs were picked out from abandoned nests of house martin swallows and identified morphologically to the species level. The bugs were randomly selected, dissected and then subjected to MALDI-TOF MS and molecular analyses. Results A total of 65 adults and 50 nymphs were used in the attempt to determine whether this tool could identify the bug species and discriminate their developmental stages. Five adults and four nymphs of C. hirundinis specimens were molecularly identified to update our MS homemade arthropod database. BLAST analysis of COI gene sequences from these C. hirundinis revealed 98.66–99.12% identity with the corresponding sequences of C. hirundinis of the GenBank. The blind test against the database supplemented with MS reference spectra showed 100% (57/57) C. hirundinis adults and 100% (46/46) C. hirundinis nymphs were reliably identified and in agreement with morphological identification with logarithmic score values between 1.922 and 2.665. Ninety-nine percent of C. hirundinis specimens tested were positive for Wolbachia spp. The sequencing results revealed that they were identical to Wolbachia massiliensis, belonging to the new T-supergroup strain and previously isolated from C. hemipterus. Conclusions We report for the first time to our knowledge a case of human infestation by swallow bugs (C. hirundinis) in France. We also show the usefulness of MALDI-TOF MS in the rapid identification of C. hirundinis specimens and nymphs with minimal sample requirements. We phylogenetically characterized the novel Wolbachia strain (W. massiliensis) infecting C. hirundinis and compared it to other recognized Wolbachia clades. Graphical Abstract

Targeted Protein Profiling of In Vivo NIPP-Treated Tissues Using DigiWest Technology

Non-invasive physical plasma (NIPP) is a novel therapeutic tool, currently being evaluated for the treatment of cancer and precancerous lesions in gynecology and other disciplines. Additionally, patients with cervical intraepithelial neoplasia (CIN) may benefit from NIPP treatment due to its non-invasive, side-effect-free, and tissue-sparing character. However, the molecular impact of in vivo NIPP treatment needs to be further investigated. For this purpose, usually only very small tissue biopsies are available after NIPP treatment. Here, we adapted DigiWest technology, a high-throughput bead-based Western blot, for the analysis of formalin-fixed paraffin-embedded (FFPE) cervical punch biopsies with a minimal sample amount. We investigated the molecular effects of NIPP treatment directly after (0 h) and 24 h after in vivo application. Results were compared to in vitro NIPP-treated human malignant cervical cells. NIPP effects were primarily based on an inhibitory impact on the cell cycle and cell growth factors. DigiWest technology was suitable for detailed protein profiling of small, primary FFPE biopsies.

Image Segmentation for Reflected-Light Microscopy: Some Theoretical Approaches

Often when we approach the study of lithologies coming from the urban environment, but in general, even from any other environment, be it a quarry, a mine, an outcrop of our interest, the first study we carry out is the one in reflected-light optical-microscopy. Reflected-light microscopy in respect to transmitted-light microscopy requires minimal sample preparation, having to polish a single surface and without the need to thin the samples to allow light to pass through them. It may be useful, already in the first analysis, to try to produce quantitative data on what we are observing. A further advantage of reflected light in an urban environment is that of being able to observe and describe the formation or interaction between opaque minerals and the environment. Information that we lose by passing directly to the transmitted light. The information that can be useful to us and that we can obtain are the relative porosity of the sample, the texture (when easily recognizable in reflected light), the maximum size and shape of the edges of the grains. To all this is added the relationship between the areas of the different crystallites identified and the possible background mass, which cannot be solved on the observation scale. When we are dealing with many samples, we do not always have the time to be able to study individually sample by sample through images, so we resort to the use of image analysis tools for image segmentation and analysis. Among these, the main thresholding method with the Otsu method, the segmentation with the k averages algorithm, and, finally, a neural network of the SOM type. In this short work, we will review the main methods of image segmentation plus an innovative method developed by our group, highlighting its strengths and weaknesses.

ATTITUDE TOWARDS MYTHS AND MISCONCEPTIONS AMONG ADOLESCENT GIRLS DURING THE MENSTRUATION

Adolescents are dened as the young people between the age groups of 10 and 19 years as per the WHO records. Adolescence is a transition period from childhood to adult life as it is regarded as the intermediate period between the childhood age and the adult age. Menstruation occurs every month periodically except during pregnancy. The maintenance of menstrual hygienic methods is a very challenging task to the adolescent girls. Social prohibitions and traditional beliefs blocked the access to get the right kind of information to adolescent girls that led to poor hygiene practices. Menstruation has often been dealt with secrecy in many cultures. Aim: Attitude towards myths and misconceptions among adolescent girls during the menstruation. To assess the attitude regarding myths Objective: and misconceptions among adolescent girls during the menstruation. To examine the level of attitude of adolescent girls towards Myths and Misconceptions Tool: The Tool used for the present study was self structured questionnaire which was developed by the researcher. Results: Many of number of adolescent girls is having low attitude i.e., having many myths and misconceptions towards menstruation and very few sample is having high attitude which is indicating that minimal sample is not having negative attitude towards the myths and misconceptions. Sample: Sample of the present study consists of 300 adolescent girls from government vocational colleges in Chittoor District of Andhra Pradesh State. There are signicant differences between age, religion, educational quali Conclusion: cations of mother's, type of family and mode of communication among adolescent girls with regard to their attitude towards myths and misconceptions during the menstruation. There is low level of attitude towards the myths and misconceptions among adolescent girls during the menstruation

Artificial Intelligence Enabled Reagent-free Imaging Hematology Analyzer

Leukocyte differential test is a widely performed clinical procedure for screening infectious diseases. Existing hematology analyzers require labor-intensive work and a panel of expensive reagents. Here we report an artificial-intelligence enabled reagent-free imaging hematology analyzer (AIRFIHA) modality that can accurately classify subpopulations of leukocytes with minimal sample preparation. AIRFIHA is realized through training a two-step residual neural network using label-free images of isolated leukocytes acquired from a custom-built quantitative phase microscope. By leveraging the rich information contained in quantitative phase images, we not only achieved high accuracy in differentiating B and T lymphocytes, but also classified CD4 and CD8 cells, therefore outperforming the classification accuracy of most current hematology analyzers. We validated the performance of AIRFIHA in a randomly selected test set and cross-validated it across all blood donors. Owing to its easy operation, low cost, and accurate discerning capability of complex leukocyte subpopulations, we envision AIRFIHA is clinically translatable and can also be deployed in resource-limited settings, e.g., during pandemic situations for the rapid screening of infectious diseases. Corresponding author(s) Email: [email protected], [email protected]