The Performance and Clinical Applicability of HER2 Digital Image Analysis in Breast Cancer: A Systematic Review.

This systematic review aims to address the research gap in the performance of computational algorithms for the digital image analysis of HER2 images in clinical settings. While numerous studies have explored various aspects of these algorithms, there is a lack of comprehensive evaluation regarding their effectiveness in real-world clinical applications. We conducted a search of the Web of Science and PubMed databases for studies published from 31 December 2013 to 30 June 2024, focusing on performance effectiveness and components such as dataset size, diversity and source, ground truth, annotation, and validation methods. The study was registered with PROSPERO (CRD42024525404). Key questions guiding this review include the following: How effective are current computational algorithms at detecting HER2 status in digital images? What are the common validation methods and dataset characteristics used in these studies? Is there standardization of algorithm evaluations of clinical applications that can improve the clinical utility and reliability of computational tools for HER2 detection in digital image analysis? We identified 6833 publications, with 25 meeting the inclusion criteria. The accuracy rate with clinical datasets varied from 84.19% to 97.9%. The highest accuracy was achieved on the publicly available Warwick dataset at 98.8% in synthesized datasets. Only 12% of studies used separate datasets for external validation; 64% of studies used a combination of accuracy, precision, recall, and F1 as a set of performance measures. Despite the high accuracy rates reported in these studies, there is a notable absence of direct evidence supporting their clinical application. To facilitate the integration of these technologies into clinical practice, there is an urgent need to address real-world challenges and overreliance on internal validation. Standardizing study designs on real clinical datasets can enhance the reliability and clinical applicability of computational algorithms in improving the detection of HER2 cancer.

Epidemiological parameters and monitoring of analysis of treatment of children and adolescents with type 1 diabetes mellitus in insulin pump therapy with modified educational program.

OBJECTIVE: Aim: The aim of the present published work is efficacy evaluation of the modified educational program in achieving the target levels of glycemia in children and adolescents with type 1 diabetes mellitus in pump insulin therapy in Almaty.. PATIENTS AND METHODS: Materials and Methods: This study involved 125 children and adolescents with type 1 DM and evaluated the effectiveness of a modified educational program at the School of Type 1 Diabetes Mellitus. Participants were divided into subgroups based on their method of glycemia evaluation. The program's effectiveness was assessed through pre-and post-training questionnaires and measurement of glycohemoglobin levels. Statistical analysis was conducted using the Statistica application. RESULTS: Results: The study evaluated the effectiveness of a modified educational program for children and adolescents with type 1 DM. The results indicated that those who participated in the modified program demonstrated significant improvements in their knowledge and ability to manage their diabetes. They were able to correctly answer 80-90% of the questionnaire questions six months to a year after the training. Additionally, those in the modified program exhibited better carbohydrate metabolism rates and achieved higher rates of their individual treatment goals, especially when using the FreeStyle Libre system for continuous monitoring of blood glucose levels. These findings suggest that a modified educational approach can significantly enhance diabetes management and treatment outcomes in children and adolescents. CONCLUSION: Conclusions: The study concluded that a modified educational program leads to better target therapy levels in children and adolescents with type 1 diabetes mellitus, highlighting the importance of motivated parents and frequent blood glycemia measurements.

Lateralized Movements during the Mating Behavior, Which Are Associated with Sex and Sexual Experience, Increase the Mating Success in Alphitobius diaperinus (Coleoptera: Tenebrionidae).

In the present study, we explored the effects of displacement directionality in mating behavior (i.e., lateralized and non-lateralized movements) on mating success (i.e., copulation occurs) and efficiency (i.e., time length at which copulation is achieved), and its association with sex and sexual experience in A. diaperinus. To do so, we carried out mating experiments and recorded the behavior of the mating pair during the whole mating sequence (i.e., precopulatory and copulatory phases). During the precopulatory phase, independently of sex and sexual experience, all beetles performed non-lateralized (i.e., backside or frontside) approaches; however, only sexually experienced beetles showed lateralized approaches (i.e., right-side and left-side). Notably, experienced males exhibited greater mating success than virgin males. After the approach, both virgin and experienced males displayed lateralized and non-lateralized mounts on the females with distinct mating success. Regardless of their sexual experience, 100% of successful mating attempts were achieved when males mounted from the females' right side. Furthermore, the development of lateralized approaches and mounts reduces the time of mating sequence span compared with non-lateralized behaviors. We highlight the importance of lateralization in mating behavior and sexual experience to achieve higher mating success, addressing a potential learning ability of beetles based on experience.

Mass Spectrometry-Based Flavor Monitoring of Peruvian Chocolate Fabrication Process.

Flavor is one of the most prominent characteristics of chocolate and is crucial in determining the price the consumer is willing to pay. At present, two types of cocoa beans have been characterized according to their flavor and aroma profile, i.e., (1) the bulk (or ordinary) and (2) the fine flavor cocoa (FFC). The FFC has been distinguished from bulk cocoa for having a great variety of flavors. Aiming to differentiate the FFC bean origin of Peruvian chocolate, an analytical methodology using gas chromatography coupled to mass spectrometry (GC-MS) was developed. This methodology allows us to characterize eleven volatile organic compounds correlated to the aromatic profile of FFC chocolate from this geographical region (based on buttery, fruity, floral, ethereal sweet, and roasted flavors). Monitoring these 11 flavor compounds during the chain of industrial processes in a retrospective way, starting from the final chocolate bar towards pre-roasted cocoa beans, allows us to better understand the cocoa flavor development involved during each stage. Hence, this methodology was useful to distinguish chocolates from different regions, north and south of Peru, and production lines. This research can benefit the chocolate industry as a quality control protocol, from the raw material to the final product.

Photophysics of structurally modified flavin derivatives in the blue-light photoreceptor YtvA: a combined experimental and theoretical study.

The light-induced processes of two flavin mononucleotide derivatives (1- and 5-deaza flavin mononucleotide, 1DFMN and 5DFMN), incorporated into the LOV domain of YtvA protein from Bacillus subtilis, were studied by a combination of experimental and computational methods. Quantum mechanics/molecular mechanics (QM/MM) calculations were carried out in which the QM part was treated by density functional theory (DFT) using the B3LYP functional for geometry optimizations and the DFT/MRCI method for spectroscopic properties, whereas the MM part was described by the CHARMM force field. 1DFMN is incorporated into the protein binding site, yielding a red-shifted absorption band (λ(max) =530 nm compared to YtvA wild-type λ(max) =445 nm), but does not undergo any LOV-typical photoreactions such as triplet and photoadduct formation. QM/MM computations confirmed the absence of a channel for triplet formation and located a radiation-free channel (through an S1/S0 conical intersection) along a hydrogen transfer path that might allow for fast deactivation. By contrast, 5DFMN-YtvA-LOV shows a blue-shifted absorption (λ(max) =410 nm) and undergoes similar photochemical processes to FMN in the wild-type protein, both with regard to the photophysics and the formation of a photoadduct with a flavin-cysteinyl covalent bond. The QM/MM calculations predict a mechanism that involves hydrogen transfer in the T1 state, followed by intersystem crossing and adduct formation in the S0 state for the forward reaction. Experimentally, in contrast to wild-type YtvA, dark-state recovery in 5DFMN-YtvA-LOV is not thermally driven but can only be accomplished after absorption of a second photon by the photoadduct, again via the triplet state. The QM/MM calculations suggest a photochemical mechanism for dark-state recovery that is accessible only for the adduct with a C4a--S bond but not for alternative adducts with a C5--S bond.

Spectroscopic and theoretical study on electronically modified chromophores in LOV domains: 8-bromo- and 8-trifluoromethyl-substituted flavins.

Two chemically synthesized flavin derivatives, 8-trifluoromethyl- and 8-bromoriboflavin (8-CF(3)RF and 8-BrRF), were photochemically characterized in H(2)O and studied spectroscopically after incorporation into the LOV domain of the blue light photoreceptor YtvA from Bacillus subtilis. The spectroscopic studies were paralleled by high-level quantum chemical calculations. In solution, 8-BrRF showed a remarkably high triplet quantum yield (0.97, parent compound riboflavin, RF: 0.6) and a small fluorescence quantum yield (0.07, RF: 0.27). For 8-CF(3)RF, the triplet yield was 0.12, and the fluorescence quantum yield was 0.7. The high triplet yield of 8-BrRF is due to the bromine heavy atom effect causing a stronger spin-orbit coupling. Theoretical calculations reveal that the decreased triplet yield of 8-CF(3)RF is due to a smaller charge transfer and a less favorable energetic position of T(2), required for intersystem crossing from S(1) to T(1), as an effect of the electron-withdrawing CF(3) group. The reconstitution of the LOV domain with the new flavins resulted in the typical LOV photochemistry, consisting of triplet state formation and covalent binding of the chromophore, followed by a thermal recovery of the parent state, albeit with different kinetics and photophysical properties.

The antibiotics roseoflavin and 8-demethyl-8-amino-riboflavin from Streptomyces davawensis are metabolized by human flavokinase and human FAD synthetase.

The non-pathogenic Gram-positive soil bacterium Streptomyces davawensis synthesizes the riboflavin (vitamin B(2)) analogs roseoflavin (RoF) and 8-demethyl-8-amino-riboflavin (AF). Both compounds are antibiotics. Notably, a number of other riboflavin analogs are currently under investigation with regard to the development of novel antiinfectives. As a first step towards understanding the metabolism of riboflavin analogs in humans, the key enzymes flavokinase (EC 2.7.1.26) and FAD synthetase (EC 2.7.7.2) were studied. Human flavokinase efficiently converted RoF and AF to roseoflavin mononucleotide (RoFMN) and 8-demethyl-8-amino-riboflavin mononucleotide (AFMN), respectively. Human FAD synthetase accepted RoFMN but not AFMN as a substrate. Consequently, roseoflavin adenine dinucleotide (RoFAD) was synthesized by the latter enzyme but not 8-demethyl-8-amino-riboflavin adenine dinucleotide (AFAD). The cofactor analogs RoFMN, AFMN and RoFAD have different physicochemical properties as compared to FMN and FAD. Thus, the cofactor analogs have the potential to render flavoenzymes inactive, which may negatively affect human metabolism. RoF, but not AF, was found to inhibit human flavokinase. In summary, we suggest that AF has a lower toxic potential and may be better suited as a lead structure to develop antimicrobial compounds.

Modulation of the photocycle of a LOV domain photoreceptor by the hydrogen-bonding network.

An extended hydrogen-bonding (HB) network stabilizes the isoalloxazine ring of the flavin mononucleotide (FMN) chromophore within the photosensing LOV domain of blue-light protein receptors, via interactions between the C(2)═O, N(3)H, C(4)═O, and N(5) groups and conserved glutamine and asparagine residues. In this work we studied the influence of the HB network on the efficiency, kinetics, and energetics of a LOV protein photocycle, involving the reversible formation of a FMN-cysteine covalent adduct. The following results were found for mutations of the conserved amino acids N94, N104, and Q123 in the Bacillus subtilis LOV protein YtvA: (i) Increased (N104D, N94D) or strongly reduced (N94A) rate of adduct formation; this latter mutation extends the lifetime of the flavin triplet state, i.e., adduct formation, more than 60-fold, from 2 µs for the wild-type (WT) protein to 129 µs. (ii) Acceleration of the overall photocycle for N94S, N94A, and Q123N, with recovery lifetimes 20, 45, and 85 times faster than for YtvA-WT, respectively. (iii) Slight modifications of FMN spectral features, correlated with the polarization of low-energy transitions. (iv) Strongly reduced (N94S) or suppressed (Q123N) structural volume changes accompanying adduct formation, as determined by optoacoustic spectroscopy. (v) Minor effects on the quantum yield, with the exception of a considerable reduction for Q123N, i.e., 0.22 vs 0.49 for YtvA-WT. The data stress the importance of the HB network in modulating the photocycle of LOV domains, while at the same time establishing a link with functional responses.

Chromophore exchange in the blue light-sensitive photoreceptor YtvA from Bacillus subtilis.

YtvA from Bacillus subtilis was found as the first prokaryotic phototropin-like blue-light-responsive photoreceptor. It is composed of two domains, the photoactive LOV (light, oxygen, voltage) domain, which binds a flavin mononucleotide (FMN) as a chromophore and a STAS (sulfate transporter/anti-sigma-factor antagonist) domain, which generates a physiological signal. Here we present a routine chromophore-exchange protocol that allows chemically synthesized, structurally modified chromophores instead of the naturally present flavin mononucleotide (FMN) chromophore to be introduced. FMN was exchanged for riboflavin (RF), flavin adenine dinucleotide (FAD), 7,8-didemethyl flavin mononucleotide (DMFMN), and 8-isopropyl flavin mononucleotide (iprFMN). LOV domains reconstituted with new flavins undergo the same photocycle as native YtvA LOV, consisting of triplet formation and covalent binding of the chromophore followed by a thermal recovery of the parent state, albeit with different kinetics and photophysical properties. Interestingly, the iprFMN chromophore, inducing steric hindrances to the protein, exhibits a very fast light-to-dark-conversion and shows a high fluorescence quantum yield (0.4). Incorporation of FAD causes an increase of its fluorescence quantum yield from 0.04 (H(2)O) to 0.2.

Excited state processes in 1-deazariboflavin studied by ultrafast fluorescence kinetics.

The photochemical reactions of the 1-deaza derivative of riboflavin (RF) have been studied by sub-ps time-resolved fluorescence spectroscopy. A potential effect of the solvent polarity and hydrogen bonding capability on the excited state (ES) dynamics was tested by measuring 1-deaza RF in water and, after acetylation of the sugar moiety, also in acetonitrile. Opposite to former reports, which indicated no fluorescence after laser excitation (Spencer et al. [1977] Biochemistry 16: 3586-3594), we find a significant fluorescence in either solvent. For both compounds, the fluorescence decays are biexponential with lifetimes in the ps time domain (1.8 and 12.5 ps for 1-deaza RF, and 3 and 87 ps for Ac(4)-1-deaza RF). In addition, a third, independent fluorescence decay was observed for both compounds. The remarkably slower second decay process for the measurements of the tetra-acetyl derivative in acetonitrile points to the involvement of hydrogen bonding or changes in the protonation/tautomerization state in the ES. In contrast to the parent compound (RF), no triplet state formation can be detected for 1-deaza RF, making the internal conversion and the fluorescence the only decay processes of the ES.

Photophysical properties of structurally and electronically modified flavin derivatives determined by spectroscopy and theoretical calculations.

Four different riboflavin (RF) derivatives, two electronically modified compounds (1- and 5-deazariboflavin, 1DRF and 5DRF) and two sterically modified compounds (7,8-didemethyl- and 8-isopropylriboflavin, DMRF and iprRF), were subjected to a combination of time-resolved measurements (absorption and fluorescence) and high-level quantum chemical investigations. Both alkyl-modified flavins showed similar fluorescence properties as the parent compound, yet 5DRF had a larger quantum yield of fluorescence (PhiF = 0.52) than RF (PhiF = 0.27). Interestingly, 1DRF did not show fluorescence at all under these steady state conditions. The triplet quantum yield was different for the modified flavins such that no triplet formation was found for 1DRF, whereas the other compounds all formed triplet states (PhiTR for 5DRF of 0.64 and 0.50 and 0.23 for iprRF and DMRF, respectively). The triplet states of the two alkyl-modified flavins decayed with similar time constants as the parent compound, whereas a shorter lifetime was measured for 5DRF (tauTR = 15 micros, compared to tauTR = 29 micros for RF). In the calculations, the flavin derivatives were modeled as lumiflavins, that is, without the ribityl chain. We conclude that for aqueous solutions of DMRF, iprRF, and 5DRF intersystem crossing (ISC) takes place from the S1 1(pipi*) to the T2 3(pipi*) state by a vibronic spin-orbit coupling mechanism, a process common to most flavins, whereas ISC is slow in excited 1DRF due to the absence of a close-by triplet state.

A glucosinolate metabolism pathway in living plant cells mediates broad-spectrum antifungal defense.

Selection pressure exerted by insects and microorganisms shapes the diversity of plant secondary metabolites. We identified a metabolic pathway for glucosinolates, known insect deterrents, that differs from the pathway activated by chewing insects. This pathway is active in living plant cells, may contribute to glucosinolate turnover, and has been recruited for broad-spectrum antifungal defense responses. The Arabidopsis CYP81F2 gene encodes a P450 monooxygenase that is essential for the pathogen-induced accumulation of 4-methoxyindol-3-ylmethylglucosinolate, which in turn is activated by the atypical PEN2 myrosinase (a type of beta-thioglucoside glucohydrolase) for antifungal defense. We propose that reiterated enzymatic cycles, controlling the generation of toxic molecules and their detoxification, enable the recruitment of glucosinolates in defense responses.