Tissue Extracts Research Articles

Molecular perspective on starfish tissue extracts: Targeting human carcinoma KB cells for anticancer therapy

The marine environment is rich in natural bioactive compounds, including zebrafish and starfish species, which have garnered attention in drug research for their remarkable tissue regeneration abilities. Zebrafish, in particular, share a genetic resemblance of around 70% with humans. We chose to investigate Luidia maculata, a starfish species, due to its remarkable tissue regeneration abilities and the potential bioactive compounds it holds, making it a promising candidate for anticancer and antioxidant therapies. Human KB carcinoma cells were subjected to L. maculata tissue extracts to assess cytotoxicity using the MTT test. Intracellular ROS levels were measured via DCFH-DA, and mitochondrial membrane potential changes were evaluated using Rh-123 staining. Oxidative DNA damage was examined with the comet test, while morphological apoptotic changes were observed through the AO/EtBr dual staining technique. The active compounds in the starfish extracts were identified using HPLC, GC–MS/MS, and FTIR analyses.The bioactive fractions extracted from L. maculata demonstrated significant anti-proliferative effects on KB carcinoma cells, inducing apoptosis. These fractions led to a notable increase in intracellular ROS levels, resulting in alterations in mitochondrial membrane potential and oxidative DNA damage in the cells. Upregulation of Bax/Caspase 3 protein expression and downregulation of Bcl-2 protein expression indicated the involvement of apoptotic pathways. Comprehensive analyses confirmed 35 starfish-derived anticancer compounds that inhibit cell proliferation, induce apoptosis, and target cancer-related pathways. Additionally, our study underscores the antioxidant potential of L. maculata starfish extracts, offering insights into marine organism-based therapies with promising medical applications.

Quantification of all 12 canonical ribonucleotides by real-time fluorogenic in vitro transcription.

Enzymatic methods to quantify deoxyribonucleoside triphosphates have existed for decades. In contrast, no general enzymatic method to quantify ribonucleoside triphosphates (rNTPs), which drive almost all cellular processes and serve as precursors of RNA, exists to date. ATP can be measured with an enzymatic luminometric method employing firefly luciferase, but the quantification of other ribonucleoside mono-, di-, and triphosphates is still a challenge for a non-specialized laboratory and practically impossible without chromatography equipment. To allow feasible quantification of ribonucleoside phosphates in any laboratory with typical molecular biology and biochemistry tools, we developed a robust microplate assay based on real-time detection of the Broccoli RNA aptamer during in vitro transcription. The assay employs the bacteriophage T7 and SP6 RNA polymerases, two oligonucleotide templates encoding the 49-nucleotide Broccoli aptamer, and a high-affinity fluorogenic aptamer-binding dye to quantify each of the four canonical rNTPs. The inclusion of nucleoside mono- and diphosphate kinases in the assay reactions enabled the quantification of the mono- and diphosphate counterparts. The assay is inherently specific and tolerates concentrated tissue and cell extracts. In summary, we describe the first chromatography-free method to quantify ATP, ADP, AMP, GTP, GDP, GMP, UTP, UDP, UMP, CTP, CDPand CMP in biological samples.

In-bag extraction of tissue through an incision in the posterior vaginal wall in laparoscopic myomectomy: a large retrospective study

BackgroundOur purpose was to describe the outcomes of transvaginal in-bag tissue extraction tissue through an incision in the posterior vaginal wall the middle part incision of posterior vagina in laparoscopic myomectomy.MethodsThis was a retrospective study of patients who received laparoscopic myomectomy and in-bag tissue extraction through an incision in the posterior vaginal wall between January 2016 and December 2022. Patient characteristics, intra- and post-operative complications, and outcomes were collected and analyzed.ResultsA total of 511women were included in the analysis. The mean largest myoma diameter was 8.44 ± 3.56 cm; mean specimen weight was 789.23 ± 276.97 g; mean operative time was 129.01 ± 53.13minutes; and mean blood loss was 175.99 ± 210.96 mL. Within 30-days of surgery, no fever, infection, or vaginal bleeding was noted in any patient, and the vaginal incisions of all patients had healed well. There were no incisional hernias, pelvic infections, and vaginal adhesions noted at follow-up 3 months after the operation. There were 37 cases of vaginal delivery of the patients after surgery, and there were no lacerations of the posterior wall vaginal incision.ConclusionsTransvaginal in-bag extraction though an incision in the posterior vaginal wall is feasible and safe for removing tissue after laparoscopic myomectomy.

Cysteine-free cone snail venom peptides: Classification of precursor proteins and identification of mature peptides.

The cysteine-free acyclic peptides present in marine cone snail venom have been much less investigated than their disulfide bonded counterparts. Precursor protein sequences derived from transcriptomic data, together with mass spectrometric fragmentation patterns for peptides present in venom duct tissue extracts, permit the identification of mature peptides. Twelve distinct gene superfamiles have been identified with precursor lengths between 64 and 158 residues. In the case of Conus monile, three distinct mature peptides have been identified, arising from two distinct protein precursors. Mature acyclic peptides are often post-translationally modified, with C-terminus amidation, a feature characteristic of neuropeptides. In the present study, 20 acyclic peptides from Conus monile and Conus betulinus were identified. The common modifications of C-terminus amidation, gamma carboxylation of glutamic acid (E to ϒ), N-terminus conversion of Gln (Q) to a pyroglutamyl residue (Z), and hydroxylation of Pro (P) to Hyp (O) are observed in one or more peptides identified in this study. Proteolytic trimming of sequences by cleavage at the C-terminus of Asn (N) residues is established. The presence of an asparagine endopeptidase is strengthened by the identification of legumain-like sequences in the transcriptome assemblies from diverse Conus species. Such sequences may be expected to have a cleavage specificity at Asn-Xxx peptide bonds.

Acute changes in the metabolome following resistance exercise combined with intake of different protein sources (cricket, pea, whey).

Separately, both exercise and protein ingestion have been shown to alter the blood and urine metabolome. This study goes a step further and examines changes in the metabolome derived from blood, urine and muscle tissue extracts in response to resistance exercise combined with ingestion of three different protein sources. In an acute parallel study, 52 young males performed one-legged resistance exercise (leg extension, 4 × 10 repetitions at 10 repetition maximum) followed by ingestion of either cricket (insect), pea or whey protein (0.25g protein/kg fat free mass). Blood and muscle tissue were collected at baseline and three hours after protein ingestion. Urine was collected at baseline and four hours after protein ingestion. Mixed-effects analyses were applied to examine the effect of the time (baseline vs. post), protein (cricket, pea, whey), and time x protein interaction. Nuclear magnetic resonance (NMR)-based metabolomics resulted in the annotation and quantification of 25 metabolites in blood, 35 in urine and 21 in muscle tissue. Changes in the muscle metabolome after combined exercise and protein intake indicated effects related to the protein source ingested. Muscle concentrations of leucine, methionine, glutamate and myo-inositol were higher after intake of whey protein compared to both cricket and pea protein. The blood metabolome revealed changes in a more ketogenic direction three hours after exercise reflecting that the trial was conducted after overnight fasting. Urinary concentration of trimethylamine N-oxide was significantly higher after ingestion of cricket than pea and whey protein. The blood, urine and muscle metabolome showed different and supplementary responses to exercise and ingestion of the different protein sources, and in synergy the summarized results provided a more complete picture of the metabolic state of the body.

BRONCHOPULMONARY DYSLASIA OF NEWBORNS AND PROSPECTS OF CELL THERAPY IN UKRAINE AND WORLDWIDE

Introduction. Premature birth is a serious challenge for modern medicine – in the world, about 14.9 million babies are born before 37 weeks of pregnancy, which is approximately 11.0% of all newborns. Bronchopulmonary dysplasia (BLD) is the most common form of chronic lung disease that affects premature babies and contributes to their morbidity and mortality. BPD develops in approximately 25.0% of infants with a birth weight of less than 1,500 g. Among infants born at less than 28 weeks' gestational age, the overall incidence of BPD is estimated to be 48–68%. The limited possibilities of BPD pharmacotherapy prompted the search for new therapeutic approaches. One of the alternatives is biological therapy based on mesenchymal stem cells (MSCs), secretome of MSCs, cryopreserved tissue extracts, etc.
 The aim of the study: to characterize current information on the therapeutic potential of mesenchymal stem cells in the treatment of newborns with bronchopulmonary dysplasia.
 Materials and methods. Publications were selected based on the databases PubMed, Clinical Key Elsevier, Cochrane Library, eBook Business Collection, and Google Scholar, which covered information about the pathogenesis of BPD and the prospects for the use of MSCs in the treatment of newborns.
 Results. Over the past decades, the understanding of the pathogenesis of BPD has evolved from a complication of secondary lung damage after mechanical ventilation and the effect of increased oxygen concentration to a multifactorial disease in which prenatal and postnatal factors are involved. Adult stem cells have been found to be naturally capable of sustaining, generating and replacing terminally differentiated cells. Stem cells function in response to physiological cell turnover or tissue damage due to trauma caused by mechanical ventilation, barotrauma, volutrauma, and hyperoxia in preterm infants. It is believed that the therapeutic effect of MSCs is related to their immunomodulatory, anti-inflammatory, angiogenic, antibacterial and regenerative effects. According to ClinicalTrials.gov, out of 301 clinical studies devoted to BPD, 18 clinical studies are currently known to study the effectiveness of MSCs in the treatment of newborns with BPD, which have already involved 770 newborns. The largest number of studies is conducted in East Asian countries (5 in China, 5 in the Republic of Korea, and 2 in Taiwan).
 Conclusions. Currently, research into the clinical therapy of BPD with stem cells is mainly focused on umbilical cord blood MSCs, umbilical cord MSCs, and bone marrow MSCs. Success in proving the safety and feasibility of incorporating stem cells for BPD led to the appearance of the biological drug PNEUMOSTEM® – a product of allogeneic MSCs obtained from human umbilical cord blood.

Microcystin profiles in European noble crayfish Astacus astacus and water in Lake Steinsfjorden, Norway

Lake Steinsfjorden, an important noble crayfish (Astacus astacus) habitat, is often affected by blooms of Planktothrix spp. that produce microcystins (MCs). A poor correlation between MCs by ELISA in the water and in crayfish tissue in a study in 2015 prompted further investigation by LC–HRMS. LC–HRMS analyses of filters from water samples and on selected crayfish tissue extracts from the 2015 study revealed the presence of known and previously unreported MCs. Crayfish samples from May and June 2015 were dominated by MCs from the Planktothrix bloom, whereas in September novel MCs that appeared to be metabolites of MC-LR were dominant, even though neither these nor MC-LR were detected in the water in 2015. A water sample from October 2016 also showed MCs typical of Planktothrix (i.e., [d-Asp3]- and [d-Asp3,Dhb7]MC-RR and -LR), but low levels of MC-RR and MC-LR were detected in the lake water for the first time. In late summer and autumn, the MC profiles of crayfish were dominated by the homonorvaline (Hnv) variant MC-LHnv, a putative metabolite of MC-LR. Taken together, ELISA, LC–HRMS and previous PCR analyses showed that although Planktothrix was part of the crayfish diet, it was not the sole source of MCs in the crayfish. Possibly, crayfish in Lake Steinsfjorden may be ingesting MCs from benthic cyanobacteria or from contaminated prey. Therefore, information on the cyanobacterial or MC content in the water column cannot safely be used to make predictions about MC concentrations in the crayfish in Lake Steinsfjorden. Interestingly, the results also show that targeted LC–MS analysis of the crayfish would at times have underestimated their MC content by nearly an order of magnitude, even if all previously reported MC variants had been included in the analysis.

Synthesis, characterization of Ca (OH)2: TiO2 nanocomposite and evaluation of its antimicrobial efficacy

Introduction and Aim: Root canal treatment is a dental operation that entails the extraction of the compromised tooth pulp tissue, sterilization of the root canal system, and subsequent filling with an inert substance. To mitigate the potential for bacterial infections and subsequent difficulties, it is customary for patients to receive a prescription for antibiotics prior to undergoing a root canal procedure. The objective of this study was to assess the effectiveness of Ca(OH)2: TiO2 nanoparticles as a potential substitute for antibiotics in root canal therapies, while also investigating their antibacterial properties against specific infections. Materials and Methods: Ca (OH)2: TiO2 nanoparticles were synthesized chemically using calcium hydroxide and varying concentrations of titanium oxide powder by a sol-gel process. The Ca (OH)2: TiO2 nanoparticles synthesized was characterized using Field Emission Scanning Electron Microscope (FE-SEM), energy dispersive X-ray (EDX), Fourier Transform Infrared Spectrometer (FT-IR), and UV-Vis spectrophotometry. The antibacterial activity of the synthesized nanoparticles was evaluated against pathogens Escherichia coli, Staphylococcus aureus, and Candida albicans spp.) by gel diffusion method. Results: FE-SEM analysis indicated that the Ca (OH)2: TiO2 composite material exhibited an amorphous structure, characterized by a particle size measuring 25.88 nm. But FTIR was focused on the spectral region between 4000 and 450 cm-1. The absorption spectra of nanoparticles composed of titanium dioxide consistently displayed a prominent peak at a wavelength of 300 nm. The results of the experiment pertaining to the biological effects of composites suggest that the inclusion of TiO2 nanoparticles at concentrations of 25%, 50%, and 75% in the composite material led to a notably wider inhibitory zone in comparison to the use of Ca(OH)2 alone, with a statistically significant p-value of 0.05. Conclusion: The use of varying concentrations of Ca (OH)2TiO2 improves microbial activity against Escherichia coli, Staphylococcus, and Candida.

Microstructural Evaluation of the Mineralized Apical Barrier Induced by a Calcium Hydroxide Paste Containing Iodoform: A Case Report

IntroductionA 65-year-old man had nonsurgical retreatment using an iodoform and calcium hydroxide paste in a maxillary left canine with persistent apical periodontitis. An apical mineralized barrier (AMB) was observed 3-months postoperatively. Unfortunately, the tooth was extracted due to a cementum tear. This provided an opportunity to analyze the AMB histologically, as there is a lack of previous reports on its microstructure. MethodsAfter extraction and removal of the granulation tissue from the root surface, the canine was processed, and observed using micro–computed tomography (μCT) and light microscopy. Thereafter, the specimen was resin-embedded specimen was evaluated by scanning electron microscopy, micro–X-ray fluorescence spectroscopy and Raman spectroscopy to understand the mechanism and nature of the AMB formation during apical healing. ResultsNonsurgical retreatment was clinically successful based on the absence of clinical symptoms of apical periodontitis and the radiographic presence of an AMB. The AMB was opaque and could be readily differentiated from dentin under a light microscope. Micro–computed tomography analysis revealed that the AMB had the same mineral density as dentin. Scanning electron microscopy revealed that the AMB had two distinct layers based on the size of the calcified particles. Elemental mapping using micro–X-ray fluorescence spectroscopy showed that the localization of calcium and phosphorus differed between AMB and other areas of biomineralization. Raman spectral mapping revealed that the surface layer of the AMB consisted of collagen, calcium carbonate, and hydroxyapatite. ConclusionsThis study explored new analytical methods for elucidating the apical wound-healing process and the nature of the mineralized repair. The findings provided detailed information on the AMB highlighting a bilaminar structure with high calcium components higher on the inside and a brightness similar to cementum not dentin and the presence of hydroxyapatite.

A novel combined technology for treating hypertrophic scars: adipose tissue extract combined with fractional CO2 laser.

Introduction: Owing to the need for liposuction and its unsuitability for allogeneic transplantation, the clinical application of stromal vascular fraction gel (SVF-gel) combined with fractional CO2 laser for scar treatment is limited. Adipose tissue extract (ATE), rich in cytokines and growth factors, offers a more convenient option for clinical practice as it can be easily prepared using purely physical methods and has low immunogenicity. We aimed to evaluate the effectiveness of ATE combined with fractional CO2 laser in the treatment of hypertrophic scars. Methods: ATE was prepared using discarded liposuction fluid from patients undergoing liposuction. A rabbit ear hypertrophic scar model was established and treated with ATE, fractional CO2 laser, or a combination. PBS was used as a control. The scar appearance and histological changes were observed. The immunohistochemistry method was used to evaluate the expression of α-SMA, while perilipin was detected using immunofluorescence. Additionally, the level of adipogenic signal C/EBPα and PPARγ mRNA was studied. Results: Following treatment, the volume of hypertrophic scar decreased, resulting in a softer texture and thinner dermis. Additionally, there was a decrease in the infiltration of inflammatory cells, and the collagen arrangement became looser and more regular, and the expression of α-SMA also decreased, with the combination of ATE and fractional laser showing the most significant improvement. Moreover, the combination group was found to promote subcutaneous fat regeneration and increase the expression of adipogenic signals C/EBPα and PPARγ. Conclusion: The combination of ATE and fractional CO2 laser treatment has been shown to inhibit the development of hypertrophic scars. This effect may be attributed to the enhancement of adipogenesis and decrease in collagen deposition.

A liquid crystal in situ gel based on rotigotine for the treatment of Parkinson's disease.

One of the most common neurodegenerative illnesses is Parkinson's disease (PD). Rotigotine (RTG) is a dopamine agonist that exerts anti-Parkinsonian effects through dopamine receptor agonism to improve motor symptoms and overall performance in PD patients. In this study, an in situ liquid crystal gel called rotigotine-gel (RTG-gel) was developed using soya phosphatidyl choline (SPC) and glycerol dioleate (GDO) to provide long-acting slow-release benefits of rotigotine while minimizing side effects. This study prepared the RTG-gel precursor solution using SPC, GDO, and ethanol (in the ratio of 54:36:10, w/w/w). The internal structures of the gel were confirmed by crossed-polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), and differential scanning calorimetry (DSC). The rheological properties of the RTG-gel precursor solution indicate a favorable combination of low viscosity and excellent flowability. The gel that produced during water absorption was also highly viscous and structurally stable, which helped to maintain the drug delayed release at the injection site. In vitro release assays showed that the in vitro release of RTG-gel followed Ritger-Peppas. The RTG-gel precursor solution was administered by subcutaneous injection, and the results of in vivo pharmacokinetic tests in SD rats showed that the plasma elimination half-life (t1/2) was 59.28 ± 16.08h; the time to peak blood concentration (Tmax) was 12.00 ± 10.32h, and the peak concentration (Cmax) was 29.9 ± 10.10ng/mL. The blood concentration remained above 0.1ng/mL for 20days after administration and was still detectable after 31days of administration, and the bioavailability of RTG can reach 72.59%. The results of in vitro solvent exchange tests showed that the RTG-gel precursor solution undergoes rapid exchange upon contact with PBS, and the diffusion of ethanol can reach 48.1% within 60min and 80% within 8h. The results of cytotoxicity test showed 89.27 ± 4.32% cell survival after administration of the drug using RTG-gel. The results of tissue extraction at the administration site showed that healing of the injection site without redness and hemorrhage could be observed after 14days of injection. The results of tissue section of the administered site showed that the inflammatory cells decreased and granulation tissue appeared after 14days of administration, and there was basically no inflammatory cell infiltration after 35days of administration, and the inflammatory reaction was basically eliminated. It shows that RTG-gel has some irritation to the injection site, but it can be recovered by itself in the later stage, and it has good biocompatibility. In summary, RTG-gel might be a potential RTG extended-release formulation for treating PD.

Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants.

Food-borne illnesses caused by pathogenic Escherichia coli strains, especially enterohaemorrhagic E. coli (EHEC), are a serious public health problem, as debilitating disease and even death from such food poisonings have been repeatedly reported. Colicin M (ColM), a non-antibiotic antimicrobial protein produced by some strains of E. coli, has shown promising activity in controlling multiple enteropathogenic strains of E. coli and related pathogens. As contaminated green leafy vegetables are a frequent source of pathogenic E. coli infections, we genetically modified (GM) two edible crops, lettuce (Lactuca sativa L.) and mizuna (Brassica rapa subsp. nipposinica var. laciniata), to stably express the ColM gene and assessed the antibacterial activity of tissue extracts from these plants against selected E. coli strains in vitro. Transgenic plants of these species were developed using Agrobacterium-mediated transformation with a vector containing the ColM-coding gene (cma) under the control of the 35S promoter. Western blot analysis of recombinant ColM protein was performed in selected transgenic plants to confirm cma gene expression and quantify ColM accumulation. Extracts of transgenic plants expressing ColM showed significant activity against two major strains of EHEC (O157:H7 and O104:H4) as well as E. coli strains resistant to beta-lactam- and carbapenem-class antibiotics. Importantly, the antibacterial activity persisted in several subsequent generations of transgenic lettuce and mizuna plants that stably expressed the ColM gene. In addition, our results also show that the antibacterial activity of dried (up to 40°C) biomass of transgenic plants remained stable without a decrease for at least three months.

In Vivo Trafficking of the Anticancer Drug Tris(8-Quinolinolato) Gallium (III) (KP46) by Gallium-68/67 PET/SPECT Imaging.

KP46 (tris(hydroxyquinolinato)gallium(III)) is an experimental, orally administered anticancer drug. Its absorption, delivery to tumours, and mode of action are poorly understood. We aimed to gain insight into these issues using gallium-67 and gallium-68 as radiotracers with SPECT and PET imaging in mice. [67Ga]KP46 and [68Ga]KP46, compared with [68Ga]gallium acetate, were used for logP measurements, in vitro cell uptake studies in A375 melanoma cells, and in vivo imaging in mice bearing A375 tumour xenografts up to 48 h after intravenous (tracer level) and oral (tracer and bulk) administration. 68Ga was more efficiently accumulated in A375 cells in vitro when presented as [68Ga]KP46 than as [68Ga]gallium acetate, but the reverse was observed when intravenously administered in vivo. After oral administration of [68/67Ga]KP46, absorption of 68Ga and 67Ga from the GI tract and delivery to tumours were poor, with the majority excreted in faeces. By 48 h, low but measurable amounts were accumulated in tumours. The distribution in tissues of absorbed radiogallium and octanol extraction of tissues suggested trafficking as free gallium rather than as KP46. We conclude that KP46 likely acts as a slow releaser of gallium ions which are inefficiently absorbed from the GI tract and trafficked to tissues, including tumour and bone.

Development of a New Enzyme-Linked Immunosorbent Assay (ELISA) for Measuring the Content of PACAP in Mammalian Tissue and Plasma.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a naturally occurring neuropeptide found in both the central and peripheral nervous systems of vertebrates. Recent studies have revealed the presence of PACAP and its corresponding receptors, namely, the pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1R), vasoactive intestinal peptide receptor 1 (VIPR1), and vasoactive intestinal peptide receptor 2 (VIPR2), in various structures implicated in migraine pathophysiology, including sensory trigeminal neurons. Human studies have demonstrated that when infused, PACAP can cause dilation of cranial vessels and result in delayed migraine-like attacks. In light of this, we present a novel ELISA assay that has been validated for quantifying PACAP in tissue extracts and human plasma. Using two well characterized antibodies specifically targeting PACAP, we successfully developed a sandwich ELISA assay, capable of detecting and accurately quantifying PACAP without any cross-reactivity to closely related peptides. The quantification range was between 5.2 pmol/L and 400 pmol/L. The recovery in plasma ranged from 98.2% to 100%. The increasing evidence pointing to the crucial role of PACAP in migraine pathophysiology necessitates the availability of tools capable of detecting changes in the circulatory levels of PACAP and its potential application as a reliable biomarker.

C-section technique vs minilaparotomy after minimally invasive uterine surgery: a retrospective cohort study

PurposeUterine leiomyomas are benign uterine tumors. The choice of surgical treatment is guided by patient's age, desire to preserve fertility or avoid "radical" surgical interventions such as hysterectomy. In laparotomy, the issue of extracting the fibroid from the cavity does not arise. However, in laparoscopy and robotic surgery, this becomes a challenge. The aim of the present study was to determine the optimal surgical approach for fibroid extraction following laparoscopic or robotic myomectomy in terms of postoperative pain, extraction time, overall surgical time, scar size, and patient satisfaction.MethodsA total of 51 patients met the inclusion criteria and were considered in our analysis: 33 patients who had undergone the "ExCITE technique" (Group A), and 18 patients a minilaparotomy procedure (Group B), after either simple myomectomy, multiple myomectomy, supracervical hysterectomy, or total hysterectomy. The diagnosis of myoma was histologically confirmed in all cases.ResultsRegarding the postoperative pain evaluation, at 6 h, patients reported 4 [3–4] vs 6 [5.3–7] on the VAS in Group A and B, as well as at 12 h, 2 [0–2] vs 3.5 [2.3–4] in Group A and B, respectively: both differences were statistically significant (p < 0.001). No statistically significant difference at 24 h from surgery was found. All patients in Group A were satisfied with the ExCITE technique, while in Group B only 67% of them. The length of the hospital stay was significantly shorter in Group A as compared to Group B (p = 0.007). In terms of the operative time for the extraction of the surgical specimen, overall operative time, and the scar size after the surgery, there was a statistically significant difference for those in Group A.ConclusionThe ExCITE technique does not require specific training and allows the surgeon to offer a minimally invasive surgical option for patients, with also an aesthetic result. It is a safe and standardized approach that ensures tissue extraction without the need for mechanical morcellation.

A Planar-Type Micro-Biopsy Tool for a Capsule-Type Endoscope Using a One-Step Nickel Electroplating Process.

Millimeter-scale biopsy tools combined with an endoscope instrument have been widely used for minimal invasive surgery and medical diagnosis. Recently, a capsule-type endoscope was developed, which requires micromachining to fabricate micro-scale biopsy tools that have a sharp tip and other complex features, e.g., nanometer-scale end-tip sharpness and a complex scalpel design. However, conventional machining approaches are not cost-effective for mass production and cannot fabricate the micrometer-scale features needed for biopsy tools. Here, we demonstrate an electroplated nickel micro-biopsy tool which features a planar shape and is suitable to be equipped with a capsule-type endoscope. Planar-type micro-biopsy tools are designed, fabricated, and evaluated through in vitro tissue dissection experiments. Various micro-biopsy tools with a long shaft and sharp tip can be easily fabricated using a thick photoresist (SU8) mold via a simple one-step lithography and nickel electroplating process. The characteristics of various micro-biopsy tool design features, including a tip taper angle, different tool geometries, and a cutting scalpel, are evaluated for efficient tissue extraction from mice intestine. These fabricated biopsy tools have shown appropriate strength and sharpness with a sufficient amount of tissue extraction for clinical applications, e.g., cancer tissue biopsy. These micro-scale biopsy tools could be easily integrated with a capsule-type endoscope and conventional forceps.

On Roth’s “human fossil” from Baradero, Buenos Aires Province, Argentina: morphological and genetic analysis

The “human fossil” from Baradero, Buenos Aires Province, Argentina, is a collection of skeleton parts first recovered by the paleontologist Santiago Roth and further studied by the anthropologist Rudolf Martin. By the end of the nineteenth century and beginning of the twentieth century it was considered one of the oldest human skeletons from South America's southern cone. Here, we present the results of an interdisciplinary approach to study and contextualize the ancient individual remains. We discuss the context of the finding by first compiling the available evidence associated with the historical information and any previous scientific publications on this individual. Then, we conducted an osteobiographical assessment, by which we evaluated the sex, age, and overall preservation of the skeleton based on morphological features. To obtain a 3D virtual reconstruction of the skull, we performed high resolution CT-scans on selected skull fragments and the mandible. This was followed by the extraction of bone tissue and tooth samples for radiocarbon and genetic analyses, which brought only limited results due to poor preservation and possible contamination. We estimate that the individual from Baradero is a middle-aged adult male. We conclude that the revision of foundational collections with current methodological tools brings new insights and clarifies long held assumptions on the significance of samples that were recovered when archaeology was not yet professionalized.

First Robot-Assisted Ivor Lewis Operation with Manual Intrapleural Anastomosis for Corrosive Esophageal Stricture

Introduction. Esophageal pathology constitutes a small share in general morbidity, however, it remains highly relevant due to its social and clinical significance. Corrosive strictures account for a large proportion among benign esophageal diseases. Treatment of the disease starts with intraluminal surgery. However, it provides only short-term effect due to the tendency of esophageal strictures to restenosis. In case of treatment failure, esophagoplasty is performed. The esophagoplasty according to Ivor Lewis with the formation of manual intrapleural anastomosis was performed in the BSMU Clinic. The abdominal and thoracic phases were performed using Da Vinci Si robotic surgical system. By the time of publication of this paper, the authors have gained experience of 15 successfully performed surgeries of this kind.Materials and methods. The paper presents a clinical case of robot-assisted esophagoplasty according to Ivor Lewis performed on patient Y., born in 1978, with the clinical diagnosis: corrosive strictures of the lower third of the esophagus with dysphagia of III–IV degree. The pathology developed as a result of surrogate alcohol consumption in September 2017. Since October 2017, the patient has been regularly treated in the Thoracic Surgery Unit of the BSMU Clinic, where she underwent a number of endoscopic surgeries, which appeared to provide a little effect. The patient underwent a subsequent surgical treatment including esophagoplasty according to Ivor Lewis with the formation of manual intrapleural anastomosis using Da Vinci Si robotic system.Results and discussion. First three days of the postoperative period, the patient remained in the Anaesthesiology and Intensive Care Unit. She received step-by-step parenteral and enteral nutrition. Control fl uoroscopic examinations revealed timely, disturbances-free evacuation and no contrast leakage from the formed anastomosis. The patient was discharged on day 11 in satisfactory condition.Conclusion. Using Da Vinci robot-assisted system in Lewis esophageal plastic surgery has a number of specific features. It provides more precise tissue extraction with preservation of stomach and esophageal vessels, moreover, mobility of the instruments enables the key phase to be performed, namely — manual suturing in the esophagoanastomosis. As a result, the risk of postoperative complications is reduced.

Non-Small Cell Lung Cancer: Targetable Variants in Concurrent Tissue and Liquid Biopsy Testing in a North Indian Cohort.

Testing for EGFR, ALK, ROS1 and MET alterations in paired tissue and plasma samples of treatment-naïve patients of NSCLC and correlating their status with overall survival. One hundred treatment-naïve patients were recruited after obtaining informed consent. Ten ml of blood was collected within a period of two weeks from histological diagnosis, prior to the start of any treatment. DNA & RNA extraction was done from formalin-fixed paraffin embedded (FFPE) tissue and total cell-free nucleic acid extraction was done from plasma samples. EGFR mutation, ALK, ROS1 and MET rearrangements were tested by ARMS (Amplification Refractory Mutation System) PCR. All statistical analyses were conducted in R version 4.1.1. A total of 61 cases showed molecular alterations in tissue samples which included EGFR mutations (47), ALK rearrangements (12), ROS1 fusion (2). MET alteration was not detected. Forty-three cases showed EGFR mutations in plasma, 26 of which were concurrently positive in tissue. Concordance observed was 62%. ALK-EML4 rearrangement, ROS1 fusion and MET were not detected in plasma samples. Sensitivity and specificity for detection of EGFR mutation in plasma were 55.3% and 67.9% respectively. Univariate Cox regression analysis showed a positive association between EGFR mutation in tissue and overall survival (HR = 0.4; 95% CI: 0.2-0.7; p = 0.003) and improved overall survival in those who received targeted therapy (HR = 0.29; 95% CI: 0.1-0.8; p = 0.02). Concurrent testing in tissue and liquid biopsy in NSCLC increased the detection of EGFR mutations (47% to 64%). This has substantial implications in deciding treatment and administration targeted therapy and the consequent overall survival.

Serum ve Plazma Örneklerinde İrisin Seviyesini Ölçmeden Önce Aprotinin İlave Etmek Gerekir mi?

Irisin is a myokine with 112 amino acids and its blood concentration is regulated by peroxisome proliferator-activated receptor-γ coactivator1-α (PGC1-α). It is released into circulation from skeletal muscle tissue after a photolytic cleavage of extracellular domain of Fibronectin type III domain-containing protein 5 (FNDC5), a type I integral membrane protein. Aprotinin is a polyvalent serin protease inhibitor. It is added to sample solutions such as serum, plasma or tissue extracts in order to inhibit serine proteases found in the sample medium. Hence, degradation of the proteins to be measured can be prevented. This study has been carried out to obtain a preliminary data if any irisin loss could be seen in the serum samples which are kept at -80°C for a long duration. For this purpose, blood samples of 10 men and 10 women volunteers aged between 25-40 has been used. Aprotinin has been added to the plasma and the serum samples have been kept at -80°C for 3 months. At the end of 3 months, irisin levels of the samples with and without aprotinin have been determined by ELISA. Statistical analysis has shown no difference between the plasma samples with or without aprotinin (p=0.525). However, a significant decrease between the serum samples with and without aprotinin (p=0.009). In conclusion, with the results of this study, no net decision could have been achieved to add aprotinin to the samples for irisin determination with ELISA in plasma and serum kept at -80°C for about 3 months.