Tissue Macromolecules Research Articles

Models of Hydration Dependent Lymphatic Opening, Interstitial Fluid Flows and Ambipolar Diffusion.

A theoretical understanding of fluid exchange and the role of initial lymph formation in tissues through mathematical/physical modeling is lacking. Here, we present three models for tissues rich in negative fixed charges due to glycosaminoglycans interacting with the extracellular matrix. We first model a lymphatic opening mechanism at relevant hydrations of the interstitium. At each hydration affecting tissue strain, two equations coupled in time are developed and solved with the new lymphatic opening and particle draining mechanism. The lymphatic opening mechanism is then included in a new model of interstitial fluid and macromolecular flow where the influence of different exclusion and available volumes for charged and neutral particles are quantified. For therapeutic interactions with cells, essential differences are found between electrically charged and neutral therapeutic substances. The interstitial fluid hydrostatic pressure gradient and flow are expressed through an extended Darcy equation, derived using similar methods as in kinetic theory of dense gases and fluid flows. Finally, a model for ambipolar diffusion of electrically charged macromolecules in tissue is developed. Our study will inform transport of charged and neutral macromolecules between the vasculature, interstitium, and the lymphatic system, thus having implications for tissue uptake of therapeutic agents.

Structural basis and molecular mechanisms of Cl- transmembrane transport in cardiomyocytes

The intracellular concentration of chlorine anions ([Cl-]i), the equilibrium potential for chlorine anions (ECl) and transmembrane chloride currents (ICl) are the factors that significantly influence the electrophysiological properties of excitable tissue, including the myocardium. Several types of chloride (anion) conductance have been identified in the heart. In recent years, a number of transmembrane proteins demonstrating chloride conductance have been identified (CFTR, ClC, TMEM16, LRRC8), and the expression of these macromolecules in cardiac tissue has been confirmed. Accumulated data allow for establishing a molecular substrate for some chloride anion currents (ICl,PKA, ICl,ir, ICl,vol, ICl,swell, ICl,Ca, Ito2) detected in the heart. Furthermore, the molecular mechanisms regulating [Cl-]i and ECl through chloride cotransporters (KCC, NKCC1) and chloride-bicarbonate exchangers have been established. The variety of structures determining chloride transmembrane conductivity and the complexity of molecular mechanisms regulating chloride homeostasis underlie the complex effects of activation of chloride transporters in the pacemaker, conduction system and working myocardium of the heart. This review discusses the structural, biophysical properties and molecular regulation of chloride transporter protein complexes identified in the myocardium. The review also covers the mechanisms by which chloride transmembrane transport influences the bioelectrical activity of cardiomyocytes.

Protective Effects of Melatonin against Carcinogen-Induced Oxidative Damage in the Thyroid.

Melatonin, primarily synthesized in the pineal gland, plays a crucial role in regulating circadian rhythms and possesses significant antioxidative properties. By neutralizing free radicals and reducing oxidative stress, melatonin emerges as a promising agent for the prevention and therapy of many different disorders, including cancer. This paper reviews the relationship between the thyroid gland and melatonin, presenting experimental evidence on the protective effects of this indoleamine against oxidative damage to macromolecules in thyroid tissue caused by documented carcinogens (as classified by the International Agency for Research on Cancer, IARC) or caused by potential carcinogens. Furthermore, the possible influence on cancer therapy in humans and the overall well-being of cancer patients are discussed. The article highlights melatonin's essential role in maintaining thyroid health and its contribution to management strategies in patients with thyroid cancer and other thyroid diseases.

Correlations of cytokines, regulating the synthesis of structural macromolecules of connective tissue, in undifferentiated connective tissue dysplasia in adolescent girls

Objective: to study the correlations of serum levels of cytokines, regulating the synthesis of structural macromolecules of connective tissue, in undifferentiated connective tissue dysplasia in adolescent girls with menstrual cycle disorders. Materials and methods: 176 adolescent girls aged 11 to 17 years with menstrual cycle disorders (MCD) and undifferentiated connective tissue dysplasia (UCTD) and 69 healthy girls of the same age were examined. Levels of cytokines IL-1β, IL-4, IL-6, IL-10, IL-17A, TNF-α, OPG, RANKL were studied in serum samples. Results: a significant increase in serum IL-6 and RANKL (p<0,01), as well as a tendency (p<0,1) to increase OPG production was found in adolescent girls with UCTD with MCD. In UCTD with NMC, correlations between the studied cytokines are much more common than in the control group (p=0,01). The most pronounced in adolescent girls with UCTD are positive correlations (moderate strength) between the concentrations of three cytokines IL-6, RANKL and OPG. Conclusions: The data obtained reflect the mechanisms of cytokine dysregulation of metabolism and remodeling of connective tissue and indicate the development of a weakly expressed systemic inflammatory process in UCTD with MCD in adolescent girls. The results of the study can be used to develop effective individualized schemes of therapeutic and preventive measures.

Non-contrast estimate of blood-brain barrier permeability in humans using arterial spin labeling and magnetization transfer at 7T.

Blood-brain barrier (BBB) dysfunction is associated with a number of central nervous system diseases. This study demonstrates the application of a novel noninvasive technique to measure the BBB permeability in the human brain at 7T. The technique exploits the fact that, when tissue macromolecules are saturated by off-resonance RF pulse, the intravascular and the extravascular (tissue) water experience different magnetization transfer effects. This principle was combined with arterial spin labeling to distinguish between the intravascular and the tissue water, and was used to calculate perfusion, water extraction fraction (E), and BBB permeability surface area product for water (PS). Simultaneous coregistered magnetization transfer ratio maps were also generated that can provide valuable additional information. Eighteen healthy volunteers (seven females), age = 27 ± 11 years and weight = 65 ± 9kg, participated in the study. Average perfusion was 67 ± 5 and 29 ± 4ml/100 g/min (p< 0.05); and E was 0.921 ± 0.025 and 0.962 ± 0.015 (p < 0.05) in the gray matter (GM) and the white matter (WM), respectively. PS was higher in the GM (171 ± 20 ml/100 g/min) compared with the WM (95 ± 18 ml/100 g/min) (p < 0.05). The parameters exhibited good reliability with test re-test experiments. The sensitivity of this technique was demonstrated by 200 mg caffeine intake, which resulted in a decrease in the resting PS by ~31%.

Sporotrichosis: A Review of a Neglected Disease in the Last 50 Years in Brazil

Sporotrichosis is caused by fungi belonging to the genus Sporothrix, which saprophytically are found in plants and organic matter. However, cats are highly susceptible to contamination with fungal spores and, when they become sick, they can transmit it to other animals and to man. The objective of this study is to carry out a systematic review on the emergency, diagnosis, clinical symptoms, therapeutics, and control of zoonotic sporotrichosis. Published data covering the last 50 years using a combination of keywords were selected to answer the question: Why has the zoonotic sporotrichosis been a neglected disease up to now? A total of 135 studies were included in this review. The studies emphasize that in recent decades, Brazil has experienced an unprecedented zoonotic outbreak of sporotrichosis. Advances on the genus Sporothrix allowed one to associate thermotolerance, capacity for melanin synthesis, potential for adhesion to tissue macromolecules, ergosterol peroxide production, and expression of virulence proteins as tools for infection and invasion in S. brasiliensis, the main species involved, although cases with S. schenckii or S. lurei were also reported. Correct diagnosis, early treatment, basic educational measures that emphasize responsible ownership of animals and reproductive control programs for felines can contribute to the control of zoonosis.

MODULATORS OF THE TRANSCRIPTION FACTORS NF-KAPA B AND NRF2 AS A MEANS RESTRICTING THE DESTRUCTION OF THE PERIODONTAL EXTRACELLULAR MATRIX IN RATS AFTER EXPERIMENTAL CRANIOCEREBRAL INJURY

This paper describes the effect of NF-κB and Nrf2 transcription factor modulators on the depolymerization of periodontal extracellular matrix biopolymers in the early post-traumatic period following the modeling of an experimental traumatic brain injury (TBI). The study was conducted on 30 white Wistar male rats weighing 180-220 g, divided into 6 groups: the 1st (pseudo-traumatized animals, control 1) the scalp of the rats was pinched with a Mikulicz clamp for one click (under ether anesthesia); the 2nd included animals after the experimental TBI modeling (control 2); rats of other groups after the TBI modeling were intraperitoneally injected with the following modulators of transcription factors for 7 days: ammonium pyrrolidine dithiocarbamate, an inhibitor of nuclear translocation of NF-κB, in a dose of 76 mg/kg, dimethyl fumarate, an inducer of the transcription factor Nrf2, in a dose of 15 mg/kg in a 10% solution of dimethyl sulfoxide, and bioflavonoids possessing NF-κB inhibitor and Nrf2 inducer properties, namely: epigallocatechin-3-gallate in a dose of 1 mg/kg and a water-soluble form of quercetin (corvitin) in a dose of 100 mg/kg (10 mg/kg in terms of quercetin). The study has shown that the TBI modeling manifests an increase in collagenolysis and depolymerization of proteoglycans and sialoglycoproteins in the periodontal soft and bone tissue at the end of the early post-traumatic period (on the 7th day) as evidenced by the growth in the concentration of free oxyproline, hexuronic acids and N-acetylneuraminic acid in the homogenate of these tissues. The administration of specific modulators of the transcription factors NF-κB and Nrf2 (ammonium pyrrolidine dithiocarbamate and dimethyl fumarate, respectively) and herbal bioflavonoids (epigallocatechin-3-gallate and quercetin) after modeling TBI significantly inhibits the processes of depolymerization of macromolecules in the connective tissue of periodontal soft and bone structures (collagen, proteoglycans, glycoproteins) with the release and increase in the concentration of their monomers (oxyproline, hexuronic and N-acetylneuraminic acids). The article compares the effectiveness of the use of specific modulators of transcription factors NF-κB and Nrf2 and bioflavonoids under experimental conditions.

Magnetization transfer imaging of ovarian cancer: initial experiences of correlation with tissue cellularity and changes following neoadjuvant chemotherapy.

Objectives:To investigate the relationship between magnetization transfer (MT) imaging and tissue macromolecules in high-grade serous ovarian cancer (HGSOC) and whether MT ratio (MTR) changes following neoadjuvant chemotherapy (NACT).Methods:This was a prospective observational study. 12 HGSOC patients were imaged before treatment. MTR was compared to quantified tissue histology and immunohistochemistry. For a subset of patients (n = 5), MT imaging was repeated after NACT. The Shapiro–Wilk test was used to assess for normality of data and Spearman’s rank-order or Pearson’s correlation tests were then used to compare MTR with tissue quantifications. The Wilcoxon signed-rank test was used to assess for changes in MTR after treatment.Results:Treatment-naïve tumour MTR was 21.9 ± 3.1% (mean ± S.D.). MTR had a positive correlation with cellularity, rho = 0.56 (p < 0.05) and a negative correlation with tumour volume, ρ = −0.72 (p = 0.01). MTR did not correlate with the extracellular proteins, collagen IV or laminin (p = 0.40 and p = 0.90). For those patients imaged before and after NACT, an increase in MTR was observed in each case with mean MTR 20.6 ± 3.1% (median 21.1) pre-treatment and 25.6 ± 3.4% (median 26.5) post-treatment (p = 0.06).Conclusion:In treatment-naïve HGSOC, MTR is associated with cellularity, possibly reflecting intracellular macromolecular concentration. MT may also detect the HGSOC response to NACT, however larger studies are required to validate this finding.Advances in knowledge:MTR in HGSOC is influenced by cellularity. This may be applied to assess for cell changes following treatment.

Reliable Assessment of Swine Renal Fibrosis Using Quantitative Magnetization Transfer Imaging.

Quantitative magnetization transfer (qMT) is useful for measurement of murine renal fibrosis at high and ultrahigh field strengths. However, its utility at clinical field strengths and in human-like kidneys remains unknown. We tested the hypothesis that qMT would successfully detect fibrosis in swine kidneys with unilateral renal artery stenosis (RAS) at 3.0 T. The qMT protocol is composed of MT scans with variable flip angles and offset frequencies, and of B0, B1, and T1 mapping. Pigs were scanned 10 weeks after RAS or control. A 2-pool model was used to fit the bound pool fraction f of the renal cortex (CO) and outer medulla (OM). Then qMT-derived f in 5 normal and 10 RAS pigs was compared with histological fibrosis determined using Masson's trichrome staining and to renal perfusion assessed with computed tomography. The qMT 2-pool model provided accurate fittings of data collected on swine kidneys. Stenotic kidneys showed significantly elevated f in both the CO (9.8% ± 2.7% vs 6.4% ± 0.9%, P = 0.002) and OM (7.6% ± 2.2% vs 4.7% ± 1.1%, P = 0.002), as compared with normal kidneys. Histology-measured renal fibrosis and qMT-derived f correlated directly in both the cortex (Pearson correlation coefficient r = 0.93, P < 0.001) and OM (r = 0.84, P = 0.002), and inversely with stenotic kidney perfusion (r = 0.85, P = 0.002). This study demonstrates the feasibility of qMT for measuring fibrosis in human-like swine kidneys, and the association between tissue macromolecule content and renal perfusion. Therefore, qMT may be useful as a tool for noninvasive assessment of renal fibrosis in subjects with RAS at clinical field strengths.

Developing a NIR emitting benzothiazolium-thioxanthene dye and its application for the design of lysosomes-targeting palladium(II) probe

‘of a near-infrared (NIR) fluorescent probe,’Palladium ions show many negative effects on human health via the interference with the tissue macromolecules and the function of enzymes. In this paper, a novel benzothiazolium-thioxanthene hemicyanine dye BTX-OH has been synthesized for the design of a near-infrared (NIR) fluorescent probe. The dye BTX-OH in aqueous solution showed its absorption and fluorescence emission maxima in the NIR region at 750 nm and 795 nm, respectively. The application of the dye in designing a Pd2+-selective fluorescence probe BTX-A based on a “protect-deprotect” strategy by using an allyl carbonate group as the specific Pd2+-responding site has been demonstrated. The probe BTX-A exhibited high selectivity and sensitivity for detecting Pd2+ ions. In the presence of Pd2+ ions, the probe showed an enhanced fluorescence response and accumulated specifically in lysosomes in living-cell imaging.

Novel corneal targeting cell penetrating peptide as an efficient nanocarrier with an effective antimicrobial activity

Delivery of therapeutics to the ocular tissues is challenging due to various anatomical and physiological barriers imposed. Cell penetrating peptides (CPPs) have emerged as potent drug nanocarriers that have been shown to overcome these barriers and enhance bioavailability of therapeutic macromolecules in deep ocular tissues. In the present study, an ocular targeting CPP has been designed by exploring potential targets of anterior ocular tissues in particular receptors, transporters and glycosaminoglycans (GAGs). The novel 11 mer peptide sequence, Corneal Targeting Sequence 1 (CorTS 1), has been developed by modifying leucine rich repeat (LRR) motif ensuring that it interacts with small leucine rich proteoglycans and collagen present in the corneal stroma. CorTS 1 exhibited dose dependent cellular translocation from 5 μM in Human Corneal Epithelial cell line (HCE) with no cytotoxicity. CorTS 1 was also found to deliver protein cargo inside HCE cells. Ex vivo tissue penetration study of CorTS 1 demonstrated in goat eyes revealed an augmented accumulation of peptide in the stromal region of cornea than in aqueous humor. Interestingly, CorTS 1 showed an antimicrobial activity against MRSA and Fusarium dimerum. Therefore, CorTS 1 can be a promising candidate with dual traits of antimicrobial agent and nanocarrier for ocular drugs.

Recent progress in mass spectrometry based molecular imaging

Imaging mass spectrometry is a rapidly developing research field, especially in the fields of life sciences and medicine. Various types of small and macromolecules in biological tissues can be scanned by imaging mass spectrometry technology. These biomolecules are closely related to the physiological and pathological states of organisms. Therefore, imaging mass spectrometry is a vital supplement to the method of optical morphological imaging. There are three techniques of imaging mass spectrometry: matrix assisted laser desorption ionization (MALDI) mass spectrometry imaging, secondary ion mass spectrometry (SIMS) imaging, and desorption electrospray ionization (DESI) mass spectrometry imaging. This review illustrated the principles and techniques of these three techniques and their applications in scientific and clinical research. The characteristics of these three types of mass spectrometry imaging can be summarized as follows: the spatial resolutions are separately 100, 10, and 0.1–1 μm; the working environments are atmospheric pressure, vacuum or low pressure, and vacuum; the analytes are small molecules, peptides or proteins or nucleic acid, small molecule or inorganic element; the application fields are environment, biomedicine, and material science.

Zinc Oxide Nanoparticles attenuate Cadmium Chloride induced tongue toxicity in adult male albino rats (Histological and Biochemical study)

Introduction: Cadmium is a toxic heavy metal naturally present in plant food and unavoidable pollutant for many dental procedures. This metal shares in toxic peroxidation and destruction of macromolecules in different tissues, besides inhibition of DNA repair enzymes. The tongue is one of the vital organs that could be affected by chronic cadmium toxicity, especially the filiform papillae and tongue skeletal muscles due to their high metabolic activity. Aim of work: to study the possible role of Zinc oxide nanoparticles (ZnO NPs) in attenuation of the structural, ultra-structural and biochemical changes in rats' tongues induced by Cadmium chloride. Materials and Methods: sixty adult male Albino rats were divided equally -according to the utilized substance- into three groups; control, Cadmium chloride (CdCl) and (CdCl+ ZnO NPs) groups; every group received the allocated materials for 4 weeks. Samples were processed for light, electron microscopic examinations and biochemical analysis (malondialdehyde; reduced glutathione; catalase; superoxide dismutase; glutathione peroxidase). Results: Cadmium produced various apparent histopathological and biochemical alterations in tongue tissues in the form of filiform papillae with blunted ends, and thick keratin plaques. Epithelial cell layers appeared with vacuolated cytoplasm and dark nuclei. Degenerated muscle fibrils with wide spaces between them and many collagen fibers. Significant decrease of tongue reduced GSH, SOD, GPx and Cat enzymes activity and significant increase in MDA level in Cadmium chloride group as compared to control group. ZnO NPs attenuated these changes and revealed pointed tipped filiform papillae, nearly normal cells and Parallel arrangements of myofibrils. The levels of reduced GSH, SOD, GPx, Cat and MDA were insignificant compared to control group. Conclusion: Zinc oxide nanoparticles (ZnO NPs) attenuated the structural and biochemical changes in rats' tongues induced by Cadmium chloride.

Acetylcholine esterase and antioxidant responses in freshwater teleost, Channa punctata exposed to chlorpyrifos and urea

We aimed to understand the toxic effects of two crop protecting agents, organophosphate pesticide, chlorpyrifos (CPF) and fertilizer, urea (U), and their binary mixtures at sublethal concentrations for 28-d in a freshwater fish Channa punctata with a battery of biochemical biomarkers in gill and liver. The study has practical value as such mixtures, so often present together in water in the agro-intensive areas, might be predicted to cause cocktail effects. Both CPF and U inhibited AChE, augmented SOD, CAT, GPx activities, and caused lipid peroxidation and depletion in tissue macromolecules in a concentration and duration-dependent manner. While U alone had less severe effects compared to CPF treatments, complex interactions were observed for three combination doses (1CPF + 1U, 2CPF + 1U, 1CPF + 2U). In their mutual effects, antagonism prevailed over other interactions when CPF and U were in equal proportion in the mixture, while synergism was observed for AchE and key antioxidant enzymes when more U was in the mixture. The present study concluded that urea in water bodies might impart adverse effects in combination with pesticides in non-target aquatic organisms such as fish, and there should be a restriction in its excessive usage.

Quantitative Magnetization Transfer Detects Renal Fibrosis in Murine Kidneys With Renal Artery Stenosis.

Renal fibrosis is a common pathway in tubulointerstitial injury and a major determinant of renal insufficiency. Collagen deposition, a key feature of renal fibrosis, may serve as an imaging biomarker to differentiate scarred from healthy kidneys. To test the feasibility of using quantitative magnetization transfer (qMT), which assesses tissue macromolecule content, to measure renal fibrosis. Prospective. Fifteen 129S1 mice were studied 4 weeks after either sham (n = 7) or unilateral renal artery stenosis (RAS, n = 8) surgeries. Magnetization transfer (MT)-weighted images were acquired at 16.4T using an MT-prepared fast-low-angle-shot sequence. Renal B0, B1, and T1 maps were also acquired, using a dual-echo gradient echo, an actual flip angle, and inversion recovery method, respectively. A two-pool model was used to estimate the bound water fraction (f) and other tissue imaging biomarkers. Masson's trichrome staining was subsequently performed ex vivo to evaluate renal fibrosis. Comparisons of renal parameters between sham and RAS were performed using independent samples t-tests. Pearson's correlation was conducted to investigate the relationship between renal fibrosis by histology and the qMT-derived bound pool fraction f. The two-pool model provided accurate fittings of measured MT signal. The qMT-derived f of RAS kidneys was significantly increased compared to sham in all kidney zones (renal cortex [CO], 7.6 ± 2.4% vs. 4.6 ± 0.6%; outer medulla [OM], 8.2 ± 4.2% vs. 4.2 ± 0.9%; inner medulla [IM] + P, 5.8 ± 1.6% vs. 2.9 ± 0.6%, all P < 0.05). Measured f correlated well with histological fibrosis in all kidney zones (CO, Pearson's correlation coefficient r = 0.95; OM, r = 0.93; IM + P, r = 0.94, all P < 0.05). The bound pool fraction f can be quantified using qMT at 16.4T in murine kidneys, increases significantly in fibrotic RAS kidneys, and correlates well with fibrosis by histology. Therefore, qMT may constitute a valuable tool for measuring renal fibrosis in RAS. 1 TECHNICAL EFFICACY STAGE: 3.

Disposition and metabolism of 2,2′-dimorpholinodiethyl ether in sprague dawley rats and B6C3F1/N mice after oral, intravenous administration, and dermal application

The specialty amine catalyst 2,2′-dimorpholinodiethyl ether (DMDEE) is a high-production volume chemical used in the production of flexible foam, high-resilient molded foam, and in coatings and adhesives. The disposition and metabolism of [14C]DMDEE (20 or 200 mg/kg) were determined in male ane female rats and mice after oral and intravenous administration and dermal application. In male and female rats, following a single oral administration, [14C]DMDEE was well-absorbed and excreted rapidly and extensively via urine (75–93%) and some in feces (∼4–8%). The total radioactivity in tissues at 24 h and 72 h (males only) following oral administration was 8–10% and ∼4%, respectively, suggesting considerable tissue distribution. A moderate amount of the total tissue radioactivity in kidney and liver were unextractable suggesting covalent binding of [14C]DMDEE-derived products in tissue macromolecules. Absorption following a single dermal application in rats was significant (∼64%) with a similar disposition pattern to oral. The oral and dermal disposition of [14C]DMDEE in male and female mice was similar to rats. Urinary products of DMDEE identified were oxidative metabolism of the morpholine ring. Coadministration of DMDEE with nitrite in rats didn’t produce the rodent carcinogen, N-nitrosomorpholine.

The Effect of a Rat Diet Without Added Cu on Redox Status in Tissues and Epigenetic Changes in the Brain

Abstract The aim of the study was to determine whether feeding rats a diet without added Cu increases oxidation of macromolecules in tissues, as well as epigenetic changes in the brain. The rats were divided into two groups: the Cu-6.5 group which was fed a diet with a standard content of Cu in mineral mixture – 6.5 mg Cu from CuCO3 per kg of diet; and the Cu-0 group which was fed a diet with a mineral mix without Cu supplementation. At the end of the experiment the rats were weighed and blood samples were collected. Finally, the rats were euthanized and then the liver, small intestine, spleen, kidneys, heart, brain, lung, testes and leg muscles were removed and weighed. In the blood of Cu-0 rats the lower Cp activity and greater GPx and CAT activity than in Cu-6.5 rats were noticed. In the liver, lungs, heart and testes of Cu-0 rats, a decreased content of Cu were noticed. Application of Cu-0 diets resulted in increased LOOH level in the small intestine, liver, and heart, as well as increased MDA content in the liver, spleen, lungs, brain and testes. The Cu-0 treatment caused a decrease in SOD activity in the heart, lungs and testes of the rats and a decrease in CAT activity in the small intestine. In the brain and testes of rats from the Cu-0 treatment, lower content of GSH + GSSG was observed. The brain of rats from the Cu-0 treatment showed an increase in the level of PCs, 8-OHdG, Casp 8 and DNA methylation. The research has shown that a deficiency of Cu in the diet impairs the body’s antioxidant defences, which in turn leads to increased lipid oxidation in the liver, small intestinal wall, heart, spleen, lungs, brain and testes, as well as to oxidation of proteins and DNA in the brain. A deficiency of Cu in the diet also increases methylation of cytosine in the brain.

Porcine Colostrum Protects the IPEC-J2 Cells and Piglet Colon Epithelium against Clostridioides (syn. Clostridium) difficile Toxin-Induced Effects

Clostridioides difficile toxins are one of the main causative agents for the clinical symptoms observed during C. difficile infection in piglets. Porcine milk has been shown to strengthen the epithelial barrier function in the piglet’s intestine and may have the potential to neutralise clostridial toxins. We hypothesised that porcine colostrum exerts protective effects against those toxins in the IPEC-J2 cells and in the colon epithelium of healthy piglets. The IPEC-J2 cells were treated with either the toxins or porcine colostrum or their combination. Analyses included measurement of trans-epithelial electrical resistance (TEER), cell viability using propidium iodide by flow cytometry, gene expression of tight junction (TJ) proteins and immune markers, immunofluorescence (IF) histology of the cytoskeleton and a TJ protein assessment. Colon tissue explants from one- and two-week-old suckling piglets and from five-week-old weaned piglets were treated with C. difficile toxins in Ussing chamber assays to assess the permeability to macromolecules (FITC-dextran, HRP), followed by analysis of gene expression of TJ proteins and immune markers. Toxins decreased viability and integrity of IPEC-J2 cells in a time-dependent manner. Porcine colostrum exerted a protective effect against toxins as indicated by TEER and IF in IPEC-J2 cells. Toxins tended to increase paracellular permeability to macromolecules in colon tissues of two-week-old piglets and downregulated gene expression of occludin in colon tissues of five-week-old piglets (p = 0.05). Porcine milk including colostrum, besides other maternal factors, may be one of the important determinants of early immune programming towards protection from C. difficile infections in the offspring.

Aphid-Triggered Changes in Oxidative Damage Markers of Nucleic Acids, Proteins, and Lipids in Maize (Zea mays L.) Seedlings

Prior experiments illustrated reactive oxygen species (ROS) overproduction in maize plants infested with bird-cherry-oat (Rhopalosiphum padi L.) aphids. However, there is no available data unveiling the impact of aphids feeding on oxidative damages of crucial macromolecules in maize tissues. Therefore, the purpose of the current study was to evaluate the scale of oxidative damages of genomic DNA, total RNA and mRNA, proteins, and lipids in seedling leaves of two maize genotypes (Złota Karłowa and Waza cvs—susceptible and relatively resistant to the aphids, respectively). The content of oxidized guanosine residues (8-hydroxy-2′-deoxyguanosine; 8-OHdG) in genomic DNA, 8-hydroxyguanosine (8-OHG) in RNA molecules, protein carbonyl groups, total thiols (T-SH), protein-bound thiols (PB-SH), non-protein thiols (NP-SH), malondialdehyde (MDA) and electrolyte leakage (EL) levels in maze plants were determined. In addition, the electrical penetration graphs (EPG) technique was used to monitor and the aphid stylet positioning and feeding modes in the hosts. Maize seedlings were infested with 0 (control), 30 or 60 R. padi adult apterae per plant. Substantial increases in the levels of RNA, protein and lipid oxidation markers in response to aphid herbivory, but no significant oxidative damages of genomic DNA, were found. Alterations in the studied parameters were dependent on maize genotype, insect abundance and infestation time.

Modeling Corticosteroid Pharmacokinetics and Pharmacodynamics, Part I: Determination and Prediction of Dexamethasone and Methylprednisolone Tissue Binding in the Rat.

The plasma and tissue binding properties of two corticosteroids, dexamethasone (DEX) and methylprednisolone (MPL), were assessed in the rat in anticipation of developing physiologically based pharmacokinetic and pharmacokinetic/pharmacodynamic models. The tissue-to-plasma partition coefficients (K P) of DEX and MPL were measured in liver, muscle, and lung in vivo after steady-state infusion and bolus injection in rats. Since K P is often governed by reversible binding to macromolecules in blood and tissue, an attempt was made to assess K P values of DEX and MPL by in vitro binding studies using rat tissue homogenates and to compare these estimates to those obtained from in vivo kinetics after dosing. The K P values of both steroids were also calculated in rat tissues using mechanistic tissue composition-based equations. The plasma binding of DEX and MPL was linear with moderate binding (60.5% and 82.5%) in male and female rats. In vivo estimates of steroid uptake appeared linear across the tested concentrations and K P was highest in liver and lowest in muscle for both steroids. Assessment of hepatic binding of MPL in vitro was severely affected by drug loss at 37°C in male liver homogenates, whereas DEX was stable in both male and female liver homogenates. With the exception of MPL in liver, in vitro-derived K P estimates reasonably agreed with in vivo values. The mechanistic equations modestly underpredicted K P for both drugs. Tissue metabolism, saturable tissue binding, and active uptake are possible factors that can complicate assessments of in vivo tissue binding of steroids when using tissue homogenates. SIGNIFICANCE STATEMENT: Assuming the free hormone hypothesis, the ratio of the unbound drug fraction in plasma and in tissues defines the tissue-to-plasma partition coefficient (K P), an important parameter in physiologically based pharmacokinetic modeling that determines total drug concentrations within tissues and the steady-state volume of distribution. This study assessed the plasma and tissue binding properties of the synthetic corticosteroids, dexamethasone and methylprednisolone, in rats using ultrafiltration and tissue homogenate techniques. In vitro-in vivo and in silico-in vivo extrapolation of K P was assessed for both drugs in liver, muscle, and lung. Although the extrapolation was fairly successful across the tissues, in vitro homogenate studies severely underpredicted the K P of methylprednisolone in liver, partly attributable to the extensive hepatic metabolism.