A Molecular Approach for Detecting Bacteria and Fungi in Healthcare Environment Aerosols: A Systematic Review.

Molecular methods have become integral to microbiological research for microbial identification. This literature review focuses on the application of molecular methods in examining airborne bacteria and fungi in healthcare facilities. In January 2024, a comprehensive electronic search was carried out in esteemed databases including PubMed, Web of Science, and Scopus, employing carefully selected keywords such as ((bacteria) OR (virus) OR (fungi)) AND (aerosol) AND ((hospital) OR (healthcare) OR (dental office)) AND ((molecular) OR (PCR) OR (NGS) OR (RNA) OR (DNA) OR (metagenomic) OR (microarray)), following the PRISMA protocol. The review specifically targets healthcare environments with elevated concentrations of pathogenic bacteria. A total of 487 articles were initially identified, but only 13 met the inclusion criteria and were included in the review. The study disclosed that the prevalent molecular methodology for appraising aerosol quality encompassed the utilization of the PCR method, incorporating either 16S rRNA (bacteria) or 18S rRNA (fungi) amplification techniques. Notably, five diverse molecular techniques, specifically PFGE, DGGE, SBT, LAMP, and DNA hybridization methods, were implemented in five distinct studies. These molecular tests exhibited superior capabilities compared to traditional bacterial and fungal cultures, providing precise strain identification. Additionally, the molecular methods allowed the detection of gene sequences associated with antibiotic resistance. In conclusion, molecular testing offers significant advantages over classical microbiological culture, providing more comprehensive information.

Optogenetic actuator - ERK biosensor circuits identify MAPK network nodes that shape ERK dynamics.

Combining single-cell measurements of ERK activity dynamics with perturbations provides insights into the MAPK network topology. We built circuits consisting of an optogenetic actuator to activate MAPK signaling and an ERK biosensor to measure single-cell ERK dynamics. This allowed us to conduct RNAi screens to investigate the role of 50 MAPK proteins in ERK dynamics. We found that the MAPK network is robust against most node perturbations. We observed that the ERK-RAF and the ERK-RSK2-SOS negative feedback operate simultaneously to regulate ERK dynamics. Bypassing the RSK2-mediated feedback, either by direct optogenetic activation of RAS, or by RSK2 perturbation, sensitized ERK dynamics to further perturbations. Similarly, targeting this feedback in a human ErbB2-dependent oncogenic signaling model increased the efficiency of a MEK inhibitor. The RSK2-mediated feedback is thus important for the ability of the MAPK network to produce consistent ERK outputs, and its perturbation can enhance the efficiency of MAPK inhibitors.

The Temporary Mental Nerve Paresthesia as an Outcome of Dentigerous Cyst Removal during Preparation for Dental Implant Placement: A Case Report.

The usage of xenograft material is widely used in almost all oral cavity bone defects for regenerative and reconstructive purposes. The presented xenograft usage in the following care report enabled good bone defect healing and enabled the preservation of affected premolars. It is quite common to use any possible variations of bone materials to ensure bone defect improved healing. In some cases, the scope of surgeries requires the removal of each cyst within close proximity to various nerves and vessels. The inferior alveolar, infraorbital, lingual, and mental nerves are those most commonly adjacent to most operating sites in jaw bones. The usage of some additional materials such as collagen sponges, bone substitutes, resorbable membranes, or other additional materials are useful in each bone defect reconstruction but should be handled with care, as described in the following case. Before planning their usage, it is important to perform each surgery with close cone beam computed tomography imaging, which is very helpful to establish the scope of each lesion and the proximity of vital structures. There are a lot of factors that might influence any possible nerve damage, especially the different nerve anatomical variations. Even factors including the subperiosteal preparation and compression of adjacent tissues might influence later nerve function. When the lesion is expanding through the buccal cortical plate and when soft tissue fluctuation is present, some special care is needed. Similar to the presented case, a limitation in crushing, blowing, or any irritation of nerve fibers improves later postoperative outcomes. When the wound and surrounding tissues are handled with care, a limited possibility of any damage or paresthesia can occur. When the nerve itself is damaged or cut, loss of function can be permanent. Immediately after or even prophylactic prescription 1-2 days before the surgery of Vitamin B with NSAIDs (Non-steroidal anti-inflammatory drugs) (or other additional supplementary medicaments can improve nerve function in time. Possible nerve damage can be divided into many etiological factors. A quite different situation arises when the nerve is pulled in by the cyst growth into the cyst wall. The presented case report describes the outcomes of a cyst removal from the mandibular basis and treatment modalities.

Review on fluoride varnishes currently recommended in dental prophylaxis.

In dentistry, fluoride compounds play a very important role in the development of teeth hard tissue. They have been modifying the development of the carious process for many years in accordance with the principles of minimally invasive therapy. Studies have confirmed their effectiveness in the prevention and treatment of carious lesions and erosion of deciduous and permanent teeth, as well as in the dentin hypersensitivity treatment. Typically, each varnish consists of 3 basic components, i.e., a resin usually in the form of mastic, shellac and/or rosin, an alcohol-based organic solvent (usually ethanol) and active agents. In the first-generation varnishes, the active agent is fluorine compounds, most often in the form of 5% NaF, while in second-generation varnishes, the composition is further enriched with calcium and phosphorus compounds in the form of CPP-ACP/CPP-ACPF, ACP, TCP, fTCP, CSPS, TMP, CXP, or CaGP. This influences the bioavailability of fluoride in the oral environment by increasing both its release from the product and its subsequent accumulation in enamel and plaque, promotes more efficient closure of dentinal tubules, and facilitates pH buffering in the oral cavity.

CODEX, a neural network approach to explore signaling dynamics landscapes.

Current studies of cell signaling dynamics that use live cell fluorescent biosensors routinely yield thousands of single-cell, heterogeneous, multi-dimensional trajectories. Typically, the extraction of relevant information from time series data relies on predefined, human-interpretable features. Without a priori knowledge of the system, the predefined features may fail to cover the entire spectrum of dynamics. Here we present CODEX, a data-driven approach based on convolutional neural networks (CNNs) that identifies patterns in time series. It does not require a priori information about the biological system and the insights into the data are built through explanations of the CNNs' predictions. CODEX provides several views of the data: visualization of all the single-cell trajectories in a low-dimensional space, identification of prototypic trajectories, and extraction of distinctive motifs. We demonstrate how CODEX can provide new insights into ERK and Akt signaling in response to various growth factors, and we recapitulate findings in p53 and TGFß-SMAD2 signaling.

Morphology of the antegonial notch and its utility in the determination of sex on skeletal materials.

A number of criteria for the assessment of biological sex, which are applied to living or recently deceased individuals, have been developed, such as genetic, chromosomal, gonadal, hormonal, and phenotypic criteria. Features of a metric and descriptive nature are used to assess the sex of skeletal materials. The diagnostic features of the skull are concentrated in the craniofacial region and around the eye sockets. The mandible is a diagnostically important part of the skull, on which a complex of features is visible. These features develop up to the third decade of life. The goal of the research was to assess the suitability of the parameters of the preangular notch, in other words, the length, height, and surface area for sex prediction applied to skeletal materials. The study included computed tomography images of the masticatory system of 194 patients, consisting of 83 females and 111 males, aged from 16 to 93 years. The three straight lines which correspond to the sides of the triangle representing the notch, in addition to its height, were determined and measured digitally. The receiver operating characteristic method was used to assess the usefulness of the studied features for the purposes of sex prediction. The sensitivity of the test ranged from 51.4% to 67% for the parameters of the preangular notch on the right-hand side, and from 44.4% to 80.2% on the left-hand side. The most reliable predictive models were obtained for two features (shown in the graphical abstract). However, when taking into account, the specificity and sensitivity of the tests presented here, only the length of side AB (a basal length of the notch) can be regarded as a feature that supports the assessment of sex on the basis of other diagnostic features of the mandible. The size and shape of the preangular notch should be treated as auxiliary features in the assessment of sex. For this reason, it is recommended that they should be applied simultaneously to the set of features described in the section on anthropological standards.

A Dimerosesquiterpene and Sesquiterpene Lactones from Artemisia argyi Inhibiting Oncogenic PI3K/AKT Signaling in Melanoma Cells.

A library of more than 2500 plant extracts was screened for activity on oncogenic signaling in melanoma cells. The ethyl acetate extract from the aerial parts of Artemisia argyi displayed pronounced inhibition of the PI3K/AKT pathway. Active compounds were tracked with the aid of HPLC-based activity profiling, and altogether 21 active compounds were isolated, including one novel dimerosequiterpenoid (1), one new disesquiterpenoid (2), three new guaianolides (3-5), 12 known sesquiterpenoids (6-17), and four known flavonoids (19-22). A new eudesmanolide derivative (13b) was isolated as an artifact formed by methanolysis. Compound 1 is the first adduct comprising a sesquiterpene lactone and a methyl jasmonate moiety. The absolute configurations of compounds 1 and 3-18 were established by comparison of their experimental and calculated ECD spectra. The absolute configuration for 2 was determined by X-ray diffraction analysis. Guaianolide 8 was the most potent sesquiterpene lactone, inhibiting the PI3K/AKT pathway with an IC50 value of 8.9 ± 0.9 µM.

Combining Activity Profiling with Advanced Annotation to Accelerate the Discovery of Natural Products Targeting Oncogenic Signaling in Melanoma.

The discovery of bioactive natural products remains a time-consuming and challenging task. The ability to link high-confidence metabolite annotations in crude extracts with activity would be highly beneficial to the drug discovery process. To address this challenge, HPLC-based activity profiling and advanced UHPLC-HRMS/MS metabolite profiling for annotation were combined to leverage the information obtained from both approaches on a crude extract scaled down to the submilligram level. This strategy was applied to a subset of an extract library screening aiming to identify natural products inhibiting oncogenic signaling in melanoma. Advanced annotation and data organization enabled the identification of compounds that were likely responsible for the activity in the extracts. These compounds belonged to two different natural product scaffolds, namely, brevipolides from a Hyptis brevipes extract and methoxylated flavonoids identified in three different extracts of Hyptis and Artemisia spp. Targeted isolation of these prioritized compounds led to five brevipolides and seven methoxylated flavonoids. Brevipolide A (1) and 6-methoxytricin (9) were the most potent compounds from each chemical class and displayed AKT activity inhibition with an IC50 of 17.6 ± 1.6 and 4.9 ± 0.2 µM, respectively.

High-Content Screening Pipeline for Natural Products Targeting Oncogenic Signaling in Melanoma.

The incidence of melanoma, the most fatal dermatological cancer, has dramatically increased over the last few decades. Modern targeted therapy with kinase inhibitors induces potent clinical responses, but drug resistance quickly develops. Combination therapy improves treatment outcomes. Therefore, novel inhibitors targeting aberrant proliferative signaling in melanoma via the MAPK/ERK and PI3K/AKT pathways are urgently needed. Biosensors were combined that report on ERK/AKT activity with image-based high-content screening and HPLC-based activity profiling. An in-house library of 2576 plant extracts was screened on two melanoma cell lines with different oncogenic mutations leading to pathological ERK/AKT activity. Out of 140 plant extract hits, 44 were selected for HPLC activity profiling. Active thymol derivatives and piperamides from Arnica montana and Piper nigrum were identified that inhibited pathological ERK and/or AKT activity. The pipeline used enabled an efficient identification of natural products targeting oncogenic signaling in melanoma.

Mining single-cell time-series datasets with Time Course Inspector.

SUMMARY: Thanks to recent advances in live cell imaging of biosensors, microscopy experiments can generate thousands of single-cell time-series. To identify sub-populations with distinct temporal behaviours that correspond to different cell fates, we developed Time Course Inspector (TCI)-a unique tool written in R/Shiny to combine time-series analysis with clustering. With TCI it is convenient to inspect time-series, plot different data views and remove outliers. TCI facilitates interactive exploration of various hierarchical clustering and cluster validation methods. We showcase TCI by analysing a single-cell signalling time-series dataset acquired using a fluorescent biosensor. AVAILABILITY AND IMPLEMENTATION: https://github.com/pertzlab/shiny-timecourse-inspector. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Nanohydroxyapatite as a Biomaterial for Peripheral Nerve Regeneration after Mechanical Damage-In Vitro Study.

Hydroxyapatite has been used in medicine for many years as a biomaterial or a cover for other biomaterials in orthopedics and dentistry. This study characterized the physicochemical properties (structure, particle size and morphology, surface properties) of Li+- and Li+/Eu3+-doped nanohydroxyapatite obtained using the wet chemistry method. The potential regenerative properties against neurite damage in cultures of neuron-like cells (SH-SY5Y and PC12 after differentiation) were also studied. The effect of nanohydroxyapatite (nHAp) on the induction of repair processes in cell cultures was assessed in tests of metabolic activity, the level of free oxygen radicals and nitric oxide, and the average length of neurites. The study showed that nanohydroxyapatite influences the increase in mitochondrial activity, which is correlated with the increase in the length of neurites. It has been shown that the doping of nanohydroxyapatite with Eu3+ ions enhances the antioxidant properties of the tested nanohydroxyapatite. These basic studies indicate its potential application in the treatment of neurite damage. These studies should be continued in primary neuronal cultures and then with in vivo models.

Temporal perturbation of ERK dynamics reveals network architecture of FGF2/MAPK signaling.

Stimulation of PC-12 cells with epidermal (EGF) versus nerve (NGF) growth factors (GFs) biases the distribution between transient and sustained single-cell ERK activity states, and between proliferation and differentiation fates within a cell population. We report that fibroblast GF (FGF2) evokes a distinct behavior that consists of a gradually changing population distribution of transient/sustained ERK signaling states in response to increasing inputs in a dose response. Temporally controlled GF perturbations of MAPK signaling dynamics applied using microfluidics reveal that this wider mix of ERK states emerges through the combination of an intracellular feedback, and competition of FGF2 binding to FGF receptors (FGFRs) and heparan sulfate proteoglycan (HSPG) co-receptors. We show that the latter experimental modality is instructive for model selection using a Bayesian parameter inference. Our results provide novel insights into how different receptor tyrosine kinase (RTK) systems differentially wire the MAPK network to fine-tune fate decisions at the cell population level.

DISORDERS OF THE INTESTINAL FLORA AND IT IS EFFECT ON SKELETAL SYSTEM DISEASES.

The intestinal microflora plays an important role in the proper functioning of human. It is complex, diverse and dynamic ecosystem that constantly strives to maintain homeostasis throughout the body. The process of acquiring intestinal microflora begins immediately after human birth. Elementary factors that determine the modification of the composition of the microflora are age, sex, diet, lifestyle. Determinate individual composition of the microflora. Microorganisms inhabiting the intestine perform various functions, from metabolic, immune to trophic. Early changes in the intestinal microflora are reflected in the state of human health. Recently, many studies have been carried out confirming the hypothesis that the bacterial flora of the gastrointestinal tract affects the normal metabolism of bone tissue, and disorders in its composition can lead to the appearance of bumps in the processes of physiological remodeling of bone tissue and contribute to the formation of many inflammations. Microorganisms that rot in the digestive tract regulate bone metabolism through three mechanisms: the impact on the immune system, the hormonal system and the impact on the absorption of minerals. Disorders in the microflora of the digestive tract can lead to the development of inflammatory bowel diseases, and as a consequence to the accelerated development of osteoporosis or arthrosis of the joints. The aim: To collect available publications confirming the impact of microflora on the skeletal system.

FAT TISSUE AND SYNTHESIZED BY HER ADIPOKINES AS MARKERS INDICATING THE DEVELOPMENT AND PROGRESS OF OSTEOARTHRITIS.

Arthrosis as osteoarthritis is a global problem that affects more and more people and is associated with severe chronic pain, reduced mobility and, consequently, disability. The etiology of degenerative disease is complex and depends on many factors. However, its course was not fully understood. One of the factors affecting the development of arthrosis is obesity. Obesity is a growing problem. Over the past 30 years, the number of overweight people has almost doubled. In people suffering from obesity, whose body mass index is above 30kg/m2, the risk of developing degenerative changes in articular cartilage is six times higher than the risk of developing this disease in people with normal body weight. Osteoarthritis is detected when the symptoms get worse where the changes are already at some stage. Therefore, a lot of research is currently underway to find suitable biomarkers, which would indicate the potential development of degenerative changes in the future and at the same time the possibility of inhibiting their activity. One of them may be adipokines, which are synthesized by adipose tissue and affect cartilage. In obese people, adipokines may contribute to the inflammation of the low charterer, whichaccompanies both obesity and arthrosis. These compounds can be specific biomarkers to assess the degree of progression and severity of osteoarthritis. The aim: To assess the importance of obesity and adipokines produced by adipose tissue as specific markers of arthrosis.

Effects of Nd:YAG laser irradiation on the growth of Candida albicans and Streptococcus mutans: in vitro study.

The purpose of this study was to evaluate the effects of Nd:YAG laser with flat-top handpiece on the in vitro growth of Candida albicans and Streptococcus mutans. The incidence of C. albicans (opportunistic commensal) and S. mutans (facultatively anaerobic) infections is increasing, despite available treatments. Cultures of Streptococcus mutans and Candida albicans were irradiated using Nd:YAG laser (LightWalker, Fotona) with flat-top handpiece (Genova, LightWalker, Fotona) at the following parameters: group G1: 0.25 W, 10 Hz, 15 s, 3 J and group G2: 1 W, 10 Hz, 60s, 59 J. The results were evaluated directly and 24 h after irradiation using a quantitative culture method (estimation of colony-forming units in 1 ml of suspension, cfu/ml), and microscopic analysis with Janus green stain and compared with control group in which laser was not applied. C. albicans was reduced by 20 up to 54% for G1, and for G2 by 10 up to 60% directly after the application. The cfu/ml values for S. mutans decreased by 13% (p = 0.1771) for G1 and 89% (p < 0.0001) for G2. In both test groups 24 h after the application, the number of colony-forming units decreased by 15-46% for G1 and by 15-64% for G2. The arrested cell division, increasing the surface area and increasing the number of metabolically inactive cells, were observed in morphometric analysis. Macroscopic and microscopic analyses revealed a reduction in cell number and a significant decrease of cell metabolism after laser application for both C. albicans and S. mutans.

Automated Comparative Metabolite Profiling of Large LC-ESIMS Data Sets in an ACD/MS Workbook Suite Add-in, and Data Clustering on a New Open-Source Web Platform FreeClust.

The technological development of LC-MS instrumentation has led to significant improvements of performance and sensitivity, enabling high-throughput analysis of complex samples, such as plant extracts. Most software suites allow preprocessing of LC-MS chromatograms to obtain comprehensive information on single constituents. However, more advanced processing needs, such as the systematic and unbiased comparative metabolite profiling of large numbers of complex LC-MS chromatograms remains a challenge. Currently, users have to rely on different tools to perform such data analyses. We developed a two-step protocol comprising a comparative metabolite profiling tool integrated in ACD/MS Workbook Suite, and a web platform developed in R language designed for clustering and visualization of chromatographic data. Initially, all relevant chromatographic and spectroscopic data (retention time, molecular ions with the respective ion abundance, and sample names) are automatically extracted and assembled in an Excel spreadsheet. The file is then loaded into an online web application that includes various statistical algorithms and provides the user with tools to compare and visualize the results in intuitive 2D heatmaps. We applied this workflow to LC-ESIMS profiles obtained from 69 honey samples. Within few hours of calculation with a standard PC, honey samples were preprocessed and organized in clusters based on their metabolite profile similarities, thereby highlighting the common metabolite patterns and distributions among samples. Implementation in the ACD/Laboratories software package enables ulterior integration of other analytical data, and in silico prediction tools for modern drug discovery.

Protective effects of levamisole, acetylsalicylic acid, and α-tocopherol against dioxin toxicity measured as the expression of AhR and COX-2 in a chicken embryo model.

Polychlorinated dibenzo-p-dioxins and dibenzofurans (dioxins) are classed as persistent organic pollutants and have adverse effects on multiple functions within the body. Dioxins are known carcinogens, immunotoxins, and teratogens. Dioxins are transformed in vivo, and interactions between the products and the aryl hydrocarbon receptor (AhR) lead to the formation of proinflammatory and toxic metabolites. The aim of this study was to determine whether α-tocopherol (vitamin E), acetylsalicylic acid (ASA), and levamisole can decrease the amount of damage caused by dioxins. Fertile Hubbard Flex commercial line chicken eggs were injected with solutions containing 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or containing TCDD and the test compounds. The chicken embryos and organs were analyzed after 7 and 13 days. The levels at which AhR and cyclooxygenase-2 (COX-2) proteins (which are induced during inflammation) were expressed were evaluated by performing immunohistochemical analyses on embryos treated with TCDD alone or with TCDD and the test compounds. TCDD caused developmental disorders and increased AhR and COX-2 expression in the chicken embryo tissues. Vitamin E, levamisole, ASA, and ASA plus vitamin E inhibited AhR and COX-2 expression in embryos after 7 days and decreased AhR and COX-2 expression in embryos after 13 days. ASA, levamisole, and ASA plus vitamin E weakened the immune response and prevented multiple organ changes. Vitamin E was not fully protective against developmental changes in the embryos.

Frequency modulation of ERK activation dynamics rewires cell fate.

Transient versus sustained ERK MAP kinase (MAPK) activation dynamics induce proliferation versus differentiation in response to epidermal (EGF) or nerve (NGF) growth factors in PC-12 cells. Duration of ERK activation has therefore been proposed to specify cell fate decisions. Using a biosensor to measure ERK activation dynamics in single living cells reveals that sustained EGF/NGF application leads to a heterogeneous mix of transient and sustained ERK activation dynamics in distinct cells of the population, different than the population average. EGF biases toward transient, while NGF biases toward sustained ERK activation responses. In contrast, pulsed growth factor application can repeatedly and homogeneously trigger ERK activity transients across the cell population. These datasets enable mathematical modeling to reveal salient features inherent to the MAPK network. Ultimately, this predicts pulsed growth factor stimulation regimes that can bypass the typical feedback activation to rewire the system toward cell differentiation irrespective of growth factor identity.

Effect of tocopherol on biochemical blood parameters in pleuritis-induced rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The aim of this study was to evaluate the effect of tocopherol on pleuritis-induced rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Rats were treated with a single TCDD dose of 5 µg/kg body weight (b.w.) and then for 3 weeks they were daily supplemented with tocopherol at a dose of 30 mg/kg b.w. The inflammation was initiated by intrapleural injection of a single dose of 1% carrageenin solution in a volume of 0.15 ml. Changes in biochemical blood parameters were measured three times at the 24th, 72nd and 120th hour of pleuritis and the blood was collected from 20 animals of each group of rats (group with the control inflammation; group treated with TCDD and with control inflammation; group treated with TCDD, supplemented with tocopherol and with the inflammation). The following biochemical parameters were measured: tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-2, IL-4, IL-6, procollagen, telopeptide, fibrinogen, cholesterol, urea, creatinine, aspartate aminotransferase (AspAT) and alanine aminotransferase (AlAT). Daily supplementation of tocopherol caused significant changes in the level of TNF, IL-1, IL-4, IL-6, urea, creatinine, AspAT and AlAT. According to the results of these studies, we suggest that tocopherol supplementation in high doses could act as a protective treatment to improve liver metabolism.

[Currently Recommended Restorative Materials in Modern Conservative Dentistry]. / Aktualnie zalecane materialy odtwórcze w nowoczesnej stomatologii zachowawczej.

Conservative treatment which restores the function, aesthetics and protects remaining tooth structure, and what is the most important, the viability of the tooth pulp, is still needed. Dental fillings replace specialized tissues of the tooth that have been lost due to caries or injury. Any decision concerning the use of a particular restorative material should be individualized and based on the competence regarding the composition, properties and characteristics of the specific restorative material. This requires continuous updating of knowledge about available dental materials as well as education of patients who, according to actual models of dental care, should be active partners in the therapeutic process. The selection of restorative materials is often related to financial abilities of the patients, and more generally to the economic model of organized health care in a particular country. Nowadays, amalgam is increasingly dislodged by adhesive materials which permit to save more tooth structure and allow to preserve natural teeth for a longer time. In the nearest future we can expect further development of minimally invasive techniques and improvements of restorative materials, especially their mechanical properties like strength and wear resistance as well as biocompatibility. The article presents restorative materials used in modern dentistry.