A novel combinatory treatment against a CDDP-resistant non-small cell lung cancer based on a Ruthenium(II)-cyclopentadienyl compound.

The therapeutic approach to many solid tumors, including non-small cell lung cancer (NSCLC), is mainly based on the use of platinum-containing anticancer agents and is often characterized by acquired or intrinsic resistance to the drug. Therefore, the search for safer and more effective drugs is still an open challenge. Two organometallic ruthenium(II)-cyclopentadienyl compounds [Ru(η5-C5H4CHO)(Me2bipy)(PPh3)]+ (RT150) and [Ru(η5-C5H4CH2OH)(Me2bipy)(PPh3)][CF3SO3] (RT151) were tested against a panel of cisplatin-resistant NSCLC cell lines and xenografts. They were more effective than cisplatin in inducing oxidative stress and DNA damage, affecting the cell cycle and causing apoptosis. Importantly, they were found to be inhibitors of drug efflux transporters. Due to this property, the compounds significantly increased the retention and cytotoxicity of cisplatin within NSCLC cells. Notably, they did not display high toxicity in vitro against non-transformed cells (red blood cells, fibroblasts, bronchial epithelial cells, cardiomyocytes, and endothelial cells). Both compounds induced vasorelaxation and reduced endothelial cell migration, suggesting potential anti-angiogenic properties. RT151 confirmed its efficacy against NSCLC xenografts resistant to cisplatin. Either alone or combined with low doses of cisplatin, RT151 showed a good biodistribution profile in the liver, kidney, spleen, lung, and tumor. Hematochemical analysis and post-mortem organ pathology confirmed the safety of the compound in vivo, also when combined with cisplatin. To sum up, we have confirmed the effectiveness of a novel class of drugs against cisplatin-resistant NSCLC. Additionally, the compounds have a good biocompatibility and safety profile.

Neurotoxicity, Neuroprotection, In Vitro MAOA/MAOB Inhibitory Activity Assessment, Molecular Docking, and Permeability Assay Studies of Newly Synthesized Hydrazones Containing a Pyrrole Ring.

Neurodegenerative diseases such as Parkinson's and Alzheimer's continue to be some of the most significant challenges in modern medicine. Recent research related to the molecular mechanisms of parkinsonism has opened up new approaches to antiparkinsonian therapy. In response to this, we present the evaluation of the potential neuroprotective and MAOA/MAOB inhibitory effects of newly synthesized hydrazones, containing a pyrrole moiety in the carboxyl fragment of the structure. The substances were studied on different brain subcellular fractions, including rat brain synaptosomes, mitochondria, and microsomes. The single application of 50 µM of each compound to the subcellular fractions showed that all substances exhibit a weak neurotoxic effect, with 7b, 7d, and 8d being the least neurotoxic representatives. The corresponding neuroprotective and antioxidant effects were also evaluated in different injury models on subcellular fractions, single out 7b, 7d, and 8d as the most prominent derivatives. A 1 µM concentration of each molecule from the series was also studied for potential hMAOA/hMAOB inhibitory effects. The results revealed a lack of hMAOA activity for all evaluated structures and the appearance of hMAOB effects, with compounds 7b, 7d, and 8d showing effects similar to those of selegiline. The best hMAOB selectivity index (>204) was determined for 7d and 8d, distinguishing these two representatives as the most promising molecules for further studies as potential selective MAOB inhibitors. The performed molecular docking simulations defined the appearance of selective MAOB inhibitory effects based on the interaction of the tested molecules with Tyr398, which is one of the components of the aromatic cage of MAOB and participated in π-π stabilization with the aromatic pyrrole ring. The preliminary PAMPA testing indicated that in relation to the blood-brain barrier (BBB) permeability, the tested pyrrole-based hydrazones may be considered as high permeable, except for 8a and 8e, which were established to be permeable in the medium range with -logP of 5.268 and 5.714, respectively, compared to the applied references.

Dufourmentel flap for surgical treatment of medium-sized congenital melanocytic nevi on the extremities.

Congenital melanocytic nevi (CMN) are quite common benign proliferations of cutaneous melanocytes. They are present at birth or arise during the first few weeks of life being upper and lower extremities one of the most common locations. To date, consistent guidelines for clinical management of CMN do not yet exist and the main reasons for removing them are medical and cosmetic. Regardless of the cause of having a CMN removed, when it comes to surgical excision of the lesion in daily practice, the single most important decision to make is how to properly close the post-excisional defect. The local Dufourmentel skin flap seems to be a reliable solution for surgical treatment of medium-sized CMN on the limbs. It takes advantage of skin laxity adjаcent to thе defect to allоw the transpositiоn of tissuе with similаr charactеristics tо the tissuе еxcisеd, which is the key for achieving good aesthetic and functional outcomes. In this brief clinical study, the author identified a group of adult patients, who had medium-sized CMN located on their extremities. The surgical technique is explained and useful tips are given. No complications and high patient satisfaction rate were registered in the series. Dufourmentel flap is a useful tool in the armamentarium of dermatologic surgery when dealing with medium-sized CMN on the extremities. Furthermore, due to its versatility this flap could also be applied for other clinical indications both benign and malignant.

In vitro toxicity evaluation of lomefloxacin-loaded MCM-41 mesoporous silica nanoparticles.

Lomefloxacin (LF) is interesting as a model molecule from a safety point of view because of its high potential for serious adverse drug effects (i.e. phototoxic reactions). In this study, MCM-41 mesoporous silica nanoparticles (MCM-41) were loaded with lomefloxacin, aiming to overcome the drug's intrinsic cytotoxicity. The good biocompatibility of the empty drug carrier (0.1-1.0 mg/ml) was established by the absence of red blood cell lysis (hemolysis assay). The cytotoxicity of empty MCM-41 and lomefloxacin-loaded MCM-41 (LF-MCM-41) was evaluated by using a battery of in vitro cytotoxicity assays: Alamar blue, lactate dehydrogenase release and reactive oxygen species formation by dichlorofluorescein assay. Three cell cultures models: hepatoma HepG2, fibroblasts L929 and endothelial EA.hy926 cells were used to compare the cytotoxicity and reactive oxygen species formation by free drug, empty MCM-41, and LF-MCM-41. The findings from the study indicated that empty MCM-41 (0.1-1.0 mg/ml) showed a low cytotoxic potential in HepG2, followed by L929 and EA.hy926 cells. Lomefloxacin loading in MCM-41 mesoporous silica nanocarrier reduced the cytotoxicity of the free lomefloxacin, especially in the high concentration (1.0 mg/ml MCM-41, containing 120 µg/ml LF). L929 and EA.hy926 cells were more sensitive to the protective effects of LF-MCM-41, compared to HepG2 cells. The results indicate that an improvement in lomefloxacin safety might be expected after incorporation in an appropriate drug delivery system.

Nonsurgical Correction of Secondary Contour Deformities of the Penile Girth.

BACKGROUND: Dealing with remaining penilе deformities following surgery to correct complications after injection of nonabsorbable substаnces tends to be troublesome. There is a lack of adequate information regarding the options for the management of such residual irregularities. In morе than 2 decades of clinical application, hyaluronic acid (HA) fillers have proved to be a safе and reliable solution fоr a great variety of age-related changes, tissue atrophy, and contour deformities. OBJECTIVES: This brief clinical study aimed to demоnstrate the author's initial еxperience with the novel use of an HA-based filler for the correction of secondary deformities of penile girth. METHODS: Rеtrospective analysis was conducted, encompassing patients who underwent correction of secondary penile deformities with HA between August 2018 and February 2020. Тhe procedures were carried оut in an ambulatory setting and a blunt cannula was used for filler infiltration. A 9-month follow-up was established. RESULTS: Five patients aged between 29 and 44 years (average, 35.8 years) were found and included in the study. The mean volumе of HA product used for the correction was 3.32 mL. An analysis of the outcomes demonstrated a high satisfaction ratе among pаtients with no complications in thе series. CONCLUSIONS: The procedure is safe, effective, and relatively simple with reproducible and reliable outcomes. Due to its excellent spreading and good tissue integration, the selected HA filler was very useful in cases where fibrosis was present because of previous surgeries.

Overexpression of a developing xylem cDNA library in transgenic poplar generates high mutation rate specific to wood formation.

We investigated feasibility of the Full-length complementary DNA OvereXpression (FOX) system as a mutagenesis approach in poplar, using developing xylem tissue. The main goal was to assess the overall mutation rate and if the system will increase instances of mutants affected in traits linked to the xylem tissue. Indeed, we found a high mutation rate of 17.7%, whereas 80% of all mutants were significantly affected in cellulose, lignin and/or hemicellulose. Cell wall biosynthesis is a major process occurring during xylem development. Enrichment of mutants affected in cell wall composition suggests that the tissue source for the FOX library influenced the occurrence of mutants affected in a trait linked to this tissue. Additionally, we found that FLcDNAs from mutants affected in cell wall composition were homologous to genes known to be involved in cell wall biosynthesis and most recovered FLcDNAs corresponded to genes whose native expression was highest in xylem. We characterized in detail a mutant line with increased diameter. The phenotype was caused by a poplar homolog of LONELY GUY 1 (LOG1), which encodes an enzyme in cytokinin biosynthesis and significantly increased xylem proliferation. The causative role of LOG1 in the observed phenotype was further reaffirmed by elevated cytokinin concentration in the mutant and recapitulation overexpression experiment wherein multiple independent lines phenocopied the original FOX mutant. Our experiments show that the FOX approach can be efficiently used for gene discovery and molecular interrogation of traits specific to woody perennial growth and development.

Safety assessment of a newly synthesized copolymer for micellar delivery of hydrophobic caffeic acid phenethyl ester.

Caffeic acid phenethyl ester (CAPE), a major pharmacologically active component of poplar type propolis, is known for its proapoptotic, anti-inflammatory, antioxidant, antiviral, and enzyme inhibiting activities. The aim of this study was to perform an in vitro and in vivo safety assessment of a micellar system based on a newly synthesized copolymer, consisting of polyglycidol and poly(allyl glycidyl ether) (C12-PAGE-PG) as a drug delivery platform for CAPE. The in vitro studies on HepG2 and L929 cells by MTT and LDH assays after treatment with the empty and CAPE-loaded micelles showed no cytotoxic effects of the empty micelles and retained cytotoxic activity of CAPE loaded in the micelles. No hemolysis or stimulation of mouse lymphocytes or macrophages was observed in vitro. In vivo hematological, biochemical, and histological assays on rats, treated with the empty (2580 and 5160 µg/kg) or CAPE-loaded (375 and 750 µg CAPE/kg) micelles did not reveal pathological changes of any of the parameters assayed after 14-days' treatment. In conclusion, initial toxicological data characterize C12-PAGE-PG as a non-toxic and promising copolymer for development of micellar drug delivery systems, particularly for a hydrophobic active substance as CAPE.

Hep G2 cell culture confluence measurement in phase-contrast micrographs - a user-friendly, open-source software-based approach.

Phase-contrast micrographs are often used for confirmation of proliferation and viability assays. However, they are usually only a qualitative tool and fail to exclude with certainty the presence of assay interference by test substances. The complexity of image analysis workflows hinders life scientists from routinely utilizing micrograph data. Here, we present an open-source software-based, combined ilastik segmentation/ImageJ measurement of area (ISIMA) approach for cell monolayer segmentation and confluence percentage measurement of phase-contrast micrographs of Hep G2 cells. The aim of this study is to test whether the proposed approach is suitable for quantitative confirmation of proliferation data, acquired by the 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Our results show that ISIMA is user-friendly and provides reproducible data, which strongly correlates to the results of the MTT assay. In conclusion, ISIMA is an affordable, simple, and fast approach for confluence quantification by researchers without image analysis background.

Dufourmentel Flap as an Aesthetic and Anatomic Refinement Tool in Upper Lip Repair.

Reconstruction of the upper lip represents a unique challenge because of its anatomical, functional and aesthetic requirements. In all the scientific sources and practical manuals the algorithms for central and lateral upper lip defects revolve around using advancement flaps, lip-switch techniques, and skin grafts only. The author offers a new approach for upper lip repair by using the Dufourmentel flap which is a pivotal local flap. The advantages and some tips are discussed.

Trauma on a Recently Augmented Breast as a Trigger for Mondor's Disease.

Mondor's disease is the eponym used to describe a self-limited phlebitis or thrombophlebitis of the superficial veins localized mainly on the thoracoabdominal area of the human body. Its clinical manifestation includes painful superficial cords causing skin retraction. This medical condition could be idiopathic, iatrogenic or a manifestation of underlying pathology such as breast cancer and seems to be more common than has been previously thought. The vast majority of the clinical studies and case reports to date focus on Mondor's disease as a disorder which is more or less directly related to a previous surgical intervention. In this case report, the author discusses the possible role of breast surgery as a predisposing factor only and the trauma on the operated breast as a trigger for onset and earlier manifestation of Mondor's disease. A special emphasis is put on the importance of trauma prevention in breast augmentation surgery, especially when maneuvers like postoperative massages are considered.Level of Evidence V This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Poplar PtabZIP1-like enhances lateral root formation and biomass growth under drought stress.

Developing drought-resistance varieties is a major goal for bioenergy crops, such as poplar (Populus), which will be grown on marginal lands with little or no water input. Root architecture can affect drought resistance, but few genes that affect root architecture in relation to water availability have been identified. Here, using activation tagging in the prime bioenergy crop poplar, we have identified a mutant that overcomes the block of lateral root (LR) formation under osmotic stress. Positioning of the tag, validation of the activation and recapitulation showed that the phenotype is caused by the poplar PtabZIP1-like (PtabZIP1L) gene with highest homology to bZIP1 from Arabidopsis. PtabZIP1L is predominantly expressed in roots, particularly in zones where lateral root primordia (LRP) initiate and LR differentiate and emerge. Transgenics overexpressing PtabZIP1L showed precocious LRP and LR development, while PtabZIP1L suppression significantly delayed both LRP and LR formation. Transgenic overexpression and suppression of PtabZIP1L also resulted in modulation of key metabolites like proline, asparagine, valine and several flavonoids. Consistently, expression of both of the poplar Proline Dehydrogenase orthologs and two of the Flavonol Synthases genes was also increased and decreased in overexpressed and suppressed transgenics, respectively. These findings suggest that PtabZIP1L mediates LR development and drought resistance through modulation of multiple metabolic pathways.

Quantitative trait locus mapping of Populus bark features and stem diameter.

BACKGROUND: Bark plays important roles in photosynthate transport and storage, along with physical and chemical protection. Bark texture varies extensively among species, from smooth to fissured to deeply furrowed, but its genetic control is unknown. This study sought to determine the main genomic regions associated with natural variation in bark features and stem diameter. Quantitative trait loci (QTL) were mapped using an interspecific pseudo-backcross pedigree (Populus trichocarpa x P. deltoides and P. deltoides) for bark texture, bark thickness and diameter collected across three years, two sites and three biological replicates per site. RESULTS: QTL specific to bark texture were highly reproducible in shared intervals across sites, years and replicates. Significant positive correlations and co-localization between trait QTL suggest pleiotropic regulators or closely linked genes. A list of candidate genes with related putative function, location close to QTL maxima and with the highest expression level in the phloem, xylem and cambium was identified. CONCLUSION: Candidate genes for bark texture included an ortholog of Arabidopsis ANAC104 (PopNAC128), which plays a role in lignified fiber cell and ray development, as well as Pinin and Fasciclin (PopFLA) genes with a role in cell adhesion, cell shape and migration. The results presented in this study provide a basis for future genomic characterization of genes found within the QTL for bark texture, bark thickness and diameter in order to better understand stem and bark development in Populus and other woody perennial plants. The QTL mapping approach identified a list of prime candidate genes for further validation using functional genomics or forward genetics approaches.

EARLY BUD-BREAK 1 (EBB1) is a regulator of release from seasonal dormancy in poplar trees.

Trees from temperate latitudes transition between growth and dormancy to survive dehydration and freezing stress during winter months. We used activation tagging to isolate a dominant mutation affecting release from dormancy and identified the corresponding gene EARLY BUD-BREAK 1 (EBB1). We demonstrate through positioning of the tag, expression analysis, and retransformation experiments that EBB1 encodes a putative APETALA2/Ethylene responsive factor transcription factor. Transgenic up-regulation of the gene caused early bud-flush, whereas down-regulation delayed bud-break. Native EBB1 expression was highest in actively growing apices, undetectable during the dormancy period, but rapidly increased before bud-break. The EBB1 transcript was localized in the L1/L2 layers of the shoot meristem and leaf primordia. EBB1-overexpressing transgenic plants displayed enlarged shoot meristems, open and poorly differentiated buds, and a higher rate of cell division in the apex. Transcriptome analyses of the EBB1 transgenics identified 971 differentially expressed genes whose expression correlated with the EBB1 expression changes in the transgenic plants. Promoter analysis among the differentially expressed genes for the presence of a canonical EBB1-binding site identified 65 putative target genes, indicative of a broad regulatory context of EBB1 function. Our results suggest that EBB1 has a major and integrative role in reactivation of meristem activity after winter dormancy.

Lower Eyelid Reconstruction After Ablation of Skin Malignancies: How Far Can We Get in a Single-Stage Procedure?

: Reconstruction of full-thickness lower eyelid defects tends to be a devastating procedure, especially when big amount of tissue has to be removed because of oncological reasons. The applied techniques are mostly difficult to execute and multistaged, often require extensive dissection, and result in scarring and facial disfigurement.The aim of the present study is to demonstrate the authors' experience in single-staged reconstructions of full-thickness defects of the lower eyelid with local tissue only. MATERIAL AND METHODS: A retrospective analysis was conducted identifying patients with postexcisional defects after skin cancer occurring in the lower eyelid, reconstructed in 1 stage. Appropriate demographic, pathological, preoperative, and postoperative clinical data and photo documentation were collected and analyzed. RESULTS: After excluding patients with partial thickness defects (n=5), a total of 13 patients with lower eyelid full-thickness defects were studied, including 9 women and 4 men with mean age 66.5 years (age range 53-79 years). The most common malignancy was basal cell carcinoma (n = 11). In one of the patients 2 synchronous neoplasms were excised simultaneously from the same lower eyelid. The reconstruction was successfully performed in 1 stage in all the patients by recruiting the same eyelid tissue and/or temporal tissue. No additional surgery was needed at a minimal follow-up of 9 months. CONCLUSION: Local tissue flaps have proven to be a reliable option for a single-stage reconstruction of large full-thickness defects. Tenzel flap technique seems to be a reliable option for repair of defects as large as 60% of the total eyelid length.

A systems biology approach identifies new regulators of poplar root development under low nitrogen.

In Populus, low nitrogen (LN) elicits rapid and vigorous lateral root (LR) proliferation, which is closely mirrored by corresponding transcriptomic changes. Using transcriptomic data, we built a genetic network encompassing a large proportion of the differentially regulated transcriptome. The network is organized in a hierarchical fashion, centered on 11 genes. Transgenic manipulations of only three of the 11 genes had a strong impact on root development under LN. These three genes encoded an F-box protein similar to Hawaiian Skirt (PtaHWS) and two transcription factors (PtaRAP2.11 and PtaNAC1). Up- and downregulation of the three genes caused increased and decreased root proliferation under LN conditions, respectively. The transgenic manipulations had a strong positive effect on growth under greenhouse conditions including increased shoot and root biomass. The three genes appeared to encompass a putative yet-unknown mechanism that underlies root development under LN. Specifically, the genes are predominantly expressed in roots and have a similar temporal response to LN. More importantly, transgenic manipulation for each of the three genes had a highly significant impact on the expression of the other two. The transgenic manipulations appear to also affect the expression of the regulatory miRNA (PtamiRNA164e) of one of the transcription factors (PtaNAC1), albeit in an opposite fashion. Consistent with a putative function of PtaHWS in proteasome degradation, treatment with proteasome inhibitor reversed the expression changes in the transgenic plants. The insights from this study will allow genetic modifications of root architecture for more efficient and dynamic nitrogen foraging in biofuel crops like poplar.

Synchronous Basal Cell Carcinoma of the Inferior Eyelid-Combined Surgical Approach for Single-Stage Ablation.

Basal cell carcinoma (BCC) is the most common cutaneous malignancy which can result in significant patient morbidity, especially when delicate periocular skin zone is affected. Late diagnosis of these tumors determines more aggressive surgical approach, which often requires combination of different reconstructive techniques and strategies. The authors report on a clinical case of a 67-year-old woman with synchronous BCC of the lower eyelid treated surgically in one stage by combining basic principles of both the aesthetic and reconstructive eyelid surgery. A special emphasis is put on the 5-step operative technique the authors have applied preserving form and function.

Invasive basal cell carcinoma in a xeroderma pigmentosum patient: facing secondary and tertiary aggressive recurrences.

Xeroderma pigmentosum (XP) is characterized by photohypersensitivity of sun-exposed tissues and several thousand-fold increased risk of developing malignant neoplasms of the skin and eyes. Inherited molecular defects in nucleotide excision repair genes cause the autosomal recessive condition XP. A 56-year-old woman with XP presented with an extensive multirecurrence basal cell carcinoma in the left naso-orbital region. At the time of the first visit, the patient had already received several interventions with local reconstructive techniques, a full course of radiotherapy, and bilateral neck dissection. A large tumor resection and free flap reconstruction were performed. Three years 9 months afterward, an aggressive recurrence occurred, and a second resection was needed. A new free flap was transferred, and microvascular anastomoses were done to the pedicle of the previously transferred flap. Nine months later, the patient returned with frontal bone tumoral lesions, and third microsurgical intervention was done. At that time, the reconstruction was practiced by a composite chimeric flap with a rib portion. Its pedicle was anastomosed to the one of the second free flaps. The objective of this article was to report the authors' experience concerning a unique case of XP requiring a complex reconstruction of the anterior skull base. Xeroderma pigmentosum patients need an early diagnosis and removal of cutaneous tumor lesions as some of them behave aggressively, especially those affecting the face. Free flaps are good solutions for reconstruction and should proceed from non-sun-exposed areas of the body. If reconstructed areas are highly radiated and/or skin tumors affect deep anatomical areas, complications are frequent.