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Arterial catheterization enables continuous hemodynamic monitoring but has been shown to cause severe complications, especially when multiple attempts are required. The aim of this study was to explore what factors were associated with multiple attempts and ultrasound use in the operating room. We performed a retrospective analysis of patients who had arterial catheters inserted at a tertiary care children’s hospital from January 2018 to March 2022, identifying clinical factors that were associated with both outcomes. A total of 3946 successful arterial catheter insertions were included. Multivariable analysis showed multiple attempts were associated with noncardiac surgery: pediatric (OR: 1.79, 95% CI: 1.30–2.51), neurologic (OR: 2.63, 95% CI: 1.89–3.57), orthopedic (OR: 3.23, 95% CI: 2.27–4.55), and non-radial artery placement (OR: 5.00, 95% CI: 3.33–7.14) (all p < 0.001). Multivariable analysis showed ultrasound use was associated with neonates (OR: 9.6, 95% CI: 4.1–22.5), infants (OR: 6.98, 95% CI: 4.67–10.42), toddlers (OR: 6.10, 95% CI: 3.8–9.8), and children (OR: 2.0, 95% CI: 1.7–2.5) compared to teenagers, with cardiac surgery being relative to other specialties—pediatric (OR: 0.48, 95% CI: 0.3–0.7), neurologic (OR: 0.27, 95% CI: 0.18–0.40), and orthopedic (OR: 0.38, 95% CI: 0.25–0.58) (all p < 0.001). In our exploratory analysis, increased odds of first-attempt arterial catheter insertion success were associated with cardiac surgery, palpation technique, and radial artery placement. Younger patient age category, ASA III and IV status, cardiac surgery, and anesthesiologist placement were associated with increased odds of ultrasound use.

Abstract. The aim of the present research is to study the efficacy of rosemary oil as an anesthetic for bighead carp (Hypophthalmichthys nobilis Richardson, 1845) and as an anesthetic used for transport of fingerlings. The fish used in the experiment have an average body weight (BW, g) of 42.13±17.90 and an average total length (TL, cm) of 16.54±2.32. Five treatments are conducted with five experimental concentrations: 0.20 ml.l-1, 0.30 ml.l-1, 0.40 ml.l-1, 0.50 ml.l-1 and 0.60 ml.l-1. For each concentration 10 fish are used or a total of 50 fish. For the transport experiment, 3 concentrations are used: 0.06 ml.l-1, 0.08 ml.l-1 and 0.10 ml.l-1. The applied stocking density is 3 fish/l. A total of 60 fish are used for each concentration or a total of 180 fish. The established results, for the first experiment, show that at concentration of 0.60 ml.l-1 the induction of anesthesia is the fastest (3.46 min). From all tested concentrations the recovery time is the shortest at the lowest concentration – 2.16 min (p≤0.001). The concentration of 0.10 ml.l-1 of rosemary oil can be used for transport of bighead carp fingerlings for a period from 1 to 2 hours. The concentration of 0.08 ml.l-1 can be applied when the duration of the transport is from 1 to 3 hours.

Abstract. The present paper investigates the body length and weight, and the size-weight variations of one-month-old European catfish reared at 4 variants of stocking density: Variant 1 – 5 spec/l; Variant 2 – 10 spec/l; Variant 3 – 15 spec/l and Variant 4 – 28 spec/l. The experiment is carried out at the Institute of Fisheries and Aquaculture, Plovdiv for a period of 16 days, using a production system consisting of tubs with continuous water flow (0.7 l/min). At the end of the experiment, the fish from each variant are sorted in three size-weight groups: A – large, B – medium and C – small. The number of fish in each group is established. From the group of the medium- and small-sized fish, 150 speciments are measured, while from the group of the large specimens, which are the smallest in number, all specimens are measured for the biometric parameters body weight (BW, g) and total body length (TL, cm). The results from the study show small variations in the length and weight of the fish reared at the lowest stocking density (Variant 1). As the density increase, the size-weight differences between the specimens from Group A also increased, while of those from Group B they decrease. The number of the medium-sized fish decrease (p≤0.001) while the number of large specimens (p≤0.01) and small fish increase (p≤0.001).

Abstract. The aim of this study is to examine the size selectivity of the pikeperch fed with one of the most widespread weed fish in fish farms, such as topmouth gudgeon (Pseudorasbora parva Temminck & Schlegel). Topmouth gudgeon (Pseudorasbora parva Temminck & Schlegel), separated in two size groups: large - group 1 (body weight of the specimens 8.63 ± 2.68 g, total length 9.58 ± 0.92 cm, and height of the body 2.06 ± 0.33 cm) and small - group 2 (body weight of the specimens 1.10 ± 0.36 g, average total body length of 5 ± 0.62 cm, and height of 0.85 ± 0.13 cm), were used for prey. The experiment was carried out under controlled laboratory conditions, in the course of 40 days, in 7 tanks. One pikeperch and 10 topmouth gudgeons from both size groups, in total 20 preys were placed in each tank. During the experimental period it was observed that pikeperch preferred to feed on smaller individuals, the difference in the number of consumed small and large fish is approximately 2 times (70 to 34, respectively).

Abstract Potassium is known for its effect on modifiable chronic diseases like hypertension, cardiac disease, diabetes (type-2), and bone health. In this study, a new method, neutron generator based neutron activation analysis (NAA), was utilized to measure potassium (K) in mouse carcasses. A DD110 neutron generator based NAA assembly was used for irradiation.Thirty-two postmortem mice (n= 16 males and 16 females, average weight $$22.02\pm 1.3$$ 22.02 ± 1.3 and $$17.9\pm 1.1$$ 17.9 ± 1.1 g) were employed for this study. Soft-tissue equivalent mouse phantoms were prepared for the calibration. All mice were irradiated for 10 minutes, and the gamma spectrum with 42K was collected using a high efficiency, high purity germanium (HPGe) detector. A lead shielding assembly was designed and developed around the HPGe detector to obtain an improved detection limit. Each mouse sample was irradiated and measured twice to reduce uncertainty. The average potassium concentration was found to be significantly higher in males $$(2846 \pm 525 \upmu g/g)$$ ( 2846 ± 525 μ g / g ) compared to females $$(2116.2 \pm 432 \upmu g/g)$$ ( 2116.2 ± 432 μ g / g ) . We also observed a significant correlation between potassium concentration and the weight of the mice. The detection limit for potassium quantification with the NAA system was 46 ppm. The radiation dose to the mouse was approximately 56 $${ \pm 1.6 }$$ ± 1.6 mSv for 10-min irradiation. In conclusion, this method is suitable for estimating individual potassium concentration in small animals. The direct evaluation of total body potassium in small animals provides a new way to estimate potassium uptake in animal models. This method can be adapted later to quantify potassium in the human hand and small animals in vivo. When used in vivo, it is also expected to be a valuable tool for longitudinal assessment, kinetics, and health outcomes.

Introduction: Immune checkpoint inhibitors have made a paradigm shift in the treatment of non-small cell lung cancer (NSCLC). However, clinical response varies widely and robust predictive biomarkers for patient stratification are lacking. Here, we characterize early on-treatment proteomic changes in blood plasma to gain a better understanding of treatment response and resistance., Methods: Pre-treatment (T0) and on-treatment (T1) plasma samples were collected from 225 NSCLC patients receiving PD-1/PD-L1 inhibitor-based regimens. Plasma was profiled using aptamer-based technology to quantify approximately 7000 plasma proteins per sample. Proteins displaying significant fold changes (T1:T0) were analyzed further to identify associations with clinical outcomes using clinical benefit and overall survival as endpoints. Bioinformatic analyses of upregulated proteins were performed to determine potential cell origins and enriched biological processes., Results: The levels of 142 proteins were significantly increased in the plasma of NSCLC patients following ICI-based treatments. Soluble PD-1 exhibited the highest increase, with a positive correlation to tumor PD-L1 status, and, in the ICI monotherapy dataset, an association with improved overall survival. Bioinformatic analysis of the ICI monotherapy dataset revealed a set of 30 upregulated proteins that formed a single, highly interconnected network, including CD8A connected to ten other proteins, suggestive of T cell activation during ICI treatment. Notably, the T cell-related network was detected regardless of clinical benefit. Lastly, circulating proteins of alveolar origin were identified as potential biomarkers of limited clinical benefit, possibly due to a link with cellular stress and lung damage., Conclusions: Our study provides insights into the biological processes activated during ICI-based therapy, highlighting the potential of plasma proteomics to identify mechanisms of therapy resistance and biomarkers for outcome., Competing Interests: EJ, GL, BY, YE, IS, ND, MH, CL, YB, MY are employed by Oncohost. RL, AD, YS are consultants of Oncohost. NR and AAm are employed by Asklepios Kliniken GmbH and Asklepios Fachkliniken Muenchen. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors declare that this study received funding from OncoHost. The funder had the following involvement in the study design, collection, analysis, interpretation of data, the writing of this article and the decision to submit it for publication., (Copyright © 2024 Bar, Leibowitz, Reinmuth, Ammendola, Jacob, Moskovitz, Levy-Barda, Lotem, Katsenelson, Agbarya, Abu-Amna, Gottfried, Harkovsky, Wolf, Tepper, Loewenthal, Yellin, Brody, Dahan, Yanko, Lahav, Harel, Raveh Shoval, Elon, Sela, Dicker and Shaked.)

Electrical current detaches bacterial biofilm from implanted instrumentation. Hypothetically, this can decrease implant-related infection and allow retention of instrumentation in cases of postoperative wound infections. We conducted a prospective animal study to investigate whether a 60-μAmp implantable direct current (DC) fusion stimulator decreases implant-related infection rates in a multilevel fixed-implant postoperative spinal wound infection model in rabbits. Three dorsal sites, T13, L3, and L6, were instrumented in each rabbit. A 60-μAmp DC fusion stimulator was implanted in a subcutaneous pouch lateral to the instrumented sites, and leads were connected to 2 of 3 sites in each rabbit. All sites were inoculated with methicillin-sensitive Staphylococcus aureus (MSSA). Rabbits were euthanized at 7 days, and cultures were obtained from the surgical sites, including wound swab, bone, and implants. No significant reduction was observed in postoperative infection rates of bone or implant with 60-μAmp DC (95% and 77%, respectively) compared with no current (91% and 82%, respectively) (P > .5). No significant difference was observed in bacterial load (Ps = .25-.72) between sites receiving DC and control sites. Currently used 60-μAmp DC implantable spinal fusion stimulators do not significantly reduce the rate of postoperative implant-related spinal wound infections in a rabbit model.

Genes encoding two ribonucleotide reductases (RNRs) were identified in members of the genus Streptomyces. One gene, nrdJ, encoded an oligomeric protein comprising four identical subunits each with a molecular mass of approximately 108 kDa. The activity of this protein depended on the presence of 5'-deoxyadenosylcobalamine (coenzyme B12), establishing it as a class II RNR. The Streptomyces clavuligerus nrdJ gene was cloned, using internal peptide sequences from the purified protein, and was found to encode a polypeptide of 961 aa. Molecular phylogenetic analysis showed that the S. clavuligerus class II RNR shares significant similarity with most other bacterial and archaeal class II RNRs. Two other genes, nrdA and nrdB, were initially identified in the Streptomyces coelicolor genome database in unannotated ORFs as encoding a class Ia RNR. Southern analysis demonstrated that the nrdAB genes were present in different Streptomyces spp. The S. coelicolor nrdAB genes were cloned and expressed in Escherichia coli, and the recombinant proteins were shown to represent a class I RNR. It was shown, using quantitative real-time PCR, that the S. clavuligerus class Ia and class II RNR genes were differentially transcribed during vegetative growth. The copy number of the class II nrdJ transcripts was approximately constant throughout the exponential phase of vegetative growth (3-5x10(5) copies per 400 ng total RNA after reverse transcription). In contrast, the copy number of the class Ia nrdAB transcripts was some 10- to 20-fold less than that of nrdJ in the early-exponential growth phase (2.8x10(4) copies), and decreased markedly at the mid-exponential (4x10(3) copies) and late-exponential phases (1.1x10(3) copies) of growth. A possible role for the involvement of two RNRs during vegetative growth is discussed.

Isopenicillin N synthase (IPNS) is a non-heme ferrous iron-dependent oxygenase that catalyzes the ring closure of delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine (ACV) to form isopenicillin N. Spectroscopic studies and the crystal structure of IPNS show that the iron atom in the active species is coordinated to two histidine and one aspartic acid residues, and to ACV, dioxygen and H2O. We previously showed by site-directed mutagenesis that residues His212, Asp214 and His268 in the IPNS of Streptomyces jumonjinensis are essential for activity and correspond to the iron ligands identified by crystallography. To evaluate the importance of the nature of the protein ligands for activity, His214 and His268 were exchanged with asparagine, aspartic acid and glutamine, and Asp214 replaced with glutamic acid, histidine and cysteine, each of which has the potential to bind iron. Only the Asp214Glu mutant retained activity, approximately 1% that of the wild type. To determine the importance of the spatial arrangement of the protein ligands for activity, His212 and His268 were separately exchanged with Asp214; both mutant enzymes were completely defective. These findings establish that IPNS activity depends critically on the presence of two histidine and one carboxylate ligands in a unique spatial arrangement within the active site. Molecular modeling studies of the active site employing the S. jumonjinensis IPNS crystal structure support this view. Measurements of iron binding by the wild type and the Asp214Glu, Asp214His and Asp214Cys-modified proteins suggest that Asp214 may have a role in catalysis as well as in iron coordination.

Isopenicillin N synthase is a key enzyme in the biosynthesis of penicillin and cephalosporin antibiotics, catalyzing the oxidative ring closure of delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine to form isopenicillin N. Recent advances in our understanding of the unique chemistry of this enzyme have come through the combined application of spectroscopic, molecular genetic and crystallographic approaches and led to important new insights into the structure and function of this enzyme. Here we review new information on the nature of the endogenous ligands that constitute the ferrous iron active site, sequence evidence for a novel structural motif involved in iron binding in this and related non-heme iron dependent dioxygenases, crystal structure studies on the enzyme and its substrate complex and the impact of these and site-directed mutagenesis studies for unraveling the mechanism of the isopenicillin N synthase reaction.

Morphogenesis in the yeast Saccharomyces cerevisiae consists primarily of bud formation. Certain cell division cycle (CDC) genes, CDC3, CDC10, CDC11, CDC12, are known to be involved in events critical to the pattern of bud growth and the completion of cytokinesis. Their products are associated with the formation of a ring of neck filaments that forms at the region of the mother cell-bud junction during mitosis. Morphogenesis in Candida albicans, a major fungal pathogen of humans, consists of both budding and the formation of hyphae. The latter is thought to be related to the pathogenesis and invasiveness of C. albicans. We have isolated and characterized C. albicans homologs of the S. cerevisiae CDC3 and CDC10 genes. Both C. albicans genes are capable of complementing defects in the respective S. cerevisiae genes. RNA analysis of one of the genes suggests that it is a regulated gene, with higher overall expression levels during the hyphal phase than in the yeast phase. Not surprisingly, DNA sequence analysis reveals that the proteins share extensive homology at the amino acid level with their respective S. cerevisiae counterparts. Related genes are also found in other species of Candida and, more importantly, in filamentous fungi such as Aspergillus nidulans and Neurospora crassa. A database search revealed significant sequence similarity with two peptides, one from Drosophila and one from mouse, suggesting strong evolutionary conservation of function.

Candida albicans is an opportunistic pathogen which may give rise to superficial and systemic infections. In the present study, C. albicans adhesion was studied by expression of C. albicans DNA sequences encoding adhesion functions in a nonadherent strain of Saccharomyces cerevisiae. Adherent transformant cells of S. cerevisiae harbouring a C. albicans genomic library cloned in a yeast-Escherichia coli shuttle vector were selected by using tissue culture-treated polystyrene as the attachment substratum. One transformant exhibited enhanced adhesion to treated and untreated polystyrene as well as autoaggregation, unlike control cells bearing the vector alone. Analysis of this clone revealed an insert of ca. 4.5 kb from C. albicans. Curing of the plasmid resulted in loss of adhesion and autoaggregation properties. A subclone bearing a reduced insert of 3.3 kb retained the ability to autoaggregate, to bind to treated and untreated polystyrene, and to adhere to buccal epithelial cells, unlike appropriate controls. Further subcloning of the insert to 2.7- and 1.9-kb fragments resulted in incremental decreases in adhesion and autoaggregation, whereas smaller fragments did not confer these properties. Hybridization of the 2.7-kb segment with C. albicans and S. cerevisiae DNA confirmed its origin as a single-copy sequence in the C. albicans genome as well as the absence of a homologous sequence in the genome of S. cerevisiae. The data suggest that the adhesion and aggregation phenomena of the transformant cells are related to expression of a C. albicans surface antigen encoded by the cloned DNA fragment.

The genes that encode thioredoxin and thioredoxin reductase of Streptomyces clavuligerus were cloned, and their DNA sequences were determined. Previously, we showed that S. clavuligerus possesses a disulfide reductase with broad substrate specificity that biochemically resembles the thioredoxin oxidoreductase system and may play a role in the biosynthesis of beta-lactam antibiotics. It consists consists of two components, a 70-kDa NADPH-dependent flavoprotein disulfide reductase with two identical subunits and a 12-kDa heat-stable protein general disulfide reductant. In this study, we found, by comparative analysis of their predicted amino acid sequences, that the 35-kDa protein is in fact thioredoxin reductase; it shares 48.7% amino acid sequence identity with Escherichia coli thioredoxin reductase, the 12-kDa protein is thioredoxin, and it shares 28 to 56% amino acid sequence identity with other thioredoxins. The streptomycete thioredoxin reductase has the identical cysteine redox-active region--Cys-Ala-Thr-Cys--and essentially the same flavin adenine dinucleotide- and NADPH dinucleotide-binding sites as E. coli thioredoxin reductase and is partially able to accept E. coli thioredoxin as a substrate. The streptomycete thioredoxin has the same cysteine redox-active segment--Trp-Cys-Gly-Pro-Cys--that is present in virtually all eucaryotic and procaryotic thioredoxins. However, in vivo it is unable to donate electrons to E. coli methionine sulfoxide reductase and does not serve as a substrate in vitro for E. coli thioredoxin reductase. The S. clavuligerus thioredoxin (trxA) and thioredoxin reductase (trxB) genes are organized in a cluster. They are transcribed in the same direction and separated by 33 nucleotides. In contrast, the trxA and trxB genes of E. coli, the only other organism in which both genes have been characterized, are physically widely separated.