Quantitative analysis of microplastics in beach sand via low-temperature solvent extraction and thermal degradation: Effects of particle size and sample depth.

Quantifying trace levels of microplastics in complex environmental media remains a challenge. In this study, an approach combining field collection of samples from different depths, sample size fractionation, and plastic quantification via pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) was employed to identify and quantify microplastics at two public beaches along the northeast coast of the U.S. (Salisbury beach, MA and Hampton beach, NH). A simple sampling tool was used to collect beach sand from depth intervals of 0-5 cm and 5-10 cm, respectively. The samples were sieved to give three size fractions: coarse (>1.2 mm), intermediate (100 µm-1.2 mm), and fine (1.2 µm-100 µm) particles. Following density separation and wet peroxide oxidation, a low-temperature solvent extraction protocol involving 2-chlorophenol was used to extract polyester (PET), polystyrene (PS), polyamide (PA), and polyvinyl chloride (PVC). The extract was analyzed using Py-GC-MS for the respective polymers, while the solid residue was pyrolyzed separately for polyethylene (PE) and polypropylene (PP). The one-step solvent extraction method significantly simplified the sample matrix and improved the sensitivity of analysis. Among the samples, PET was detected in greater quantities in the fine fraction than in the intermediate size fraction, and PET fine particles were located predominantly in the surface sand. Similar to PET, PS was detected at higher mass concentrations in the fine particles in most samples. These results underscore the importance of beach environment for plastic fragmentation, where a combination of factors including UV irradiation, mechanical abrasion, and water exposure promote plastic breakdown. Surface accumulation of fine plastic particles may also be attributed to transport of microplastics through wind and tides. The proposed sample treatment and analysis methods may allow sensitive and quantitative measurements of size or depth-related distribution patterns of microplastics in complex environmental media.

Vascularized hemi-hamate graft: Anatomic description of a novel pedicled osteo-chondro-ligamentous flap for proximal scaphoid reconstruction.

Scaphoid proximal pole destruction remains a surgical challenge owing to its high propensity for nonunion and osteonecrosis. The hemi-hamate graft has shown promising results in addressing this issue. However, long-term results of non-vascularized composite grafts remain uncertain. The purpose of this study was to investigate the feasibility of a vascularized hemi-hamate osteo-chondro-ligamentous pedicled flap for the reconstruction of the proximal pole of the scaphoid. Thirty fresh cadaveric wrists were used to harvest the hamate proximal pole on the dorsal intercarpal arch. A loss of substance of the scaphoid proximal pole was simulated and the hamate flap was transferred. In 15 wrists, a canulated screw osteosynthesis was performed to assess donor site morbidity and carpus stability on post-osteosynthesis dynamic radiographs. This study suggests that the proximal hamate can be harvested pedicled on the dorsal intercarpal arch. The pedicle (average pedicle diameter 0.9 mm, mean length 31.5 mm) allowed tension-free graft placement in all dissections, except for one. The morphology of the graft was very similar to that of the scaphoid proximal pole and the palmar capito-hamate ligament allowed scapholunate ligament reconstruction in all dissections. This is the first study that describes the use of a pedicled flap to fully reconstruct the complex osteo-chondro-ligamentous anatomy of the scaphoid proximal pole. This vascularized hemi-hamate flap could facilitate better long-term preservation of cartilage biomechanical properties compared to non-vascularized grafts. Donor site morbidity requires further investigation before recommending clinical use.

Study of phenanthrenes from their unique mass spectrometric behavior through quantum chemical calculations to liquid chromatographic quantitation.

Phenanthrenes and their derivatives have biological relevance owing to their antimicrobial, antioxidant, and cytotoxic effects on cancer cells. They can be efficiently analyzed through ultrahigh-performance liquid chromatography coupled to tandem high-resolution mass spectrometry (UHPLC-MS/HRMS). Herein, we first studied the unique fragmentation behavior of phenanthrenes based on direct infusion MS/HRMS analysis. As a newly described phenomenon, "organ pipe distribution", we found a structural connection linking their unique fragmentation pattern to serial H radical losses. The bonds responsible for this behavior were identified through quantum chemical calculations using a stepwise approach. Furthermore, the chromatographic aspect of this study was enhanced by developing, validating, and applying a new unscheduled targeted UHPLC-MS/HRMS method for quantifying phenanthrenes in Juncus compressus herb. Targeted compounds were efficiently separated within 4 min upon utilizing the Accucore C30 column, and the unscheduled targeted analytical approach afforded five new isomers. Compounds 1 (effususol), 3 (dehydroeffusol), and 6 (7-hydroxy-1-methyl-2-methoxy-5-vinyl-9,10-dihydrophenanthrene) had their linearity limits determined within 10-5000 nM, and Compounds 2 (effusol), 4 (juncusol), 5 (effususin A), and 7 (compressin A) within 25-5000 nM. The coefficients of variation for precision ranged from 1.4 % to 15.2 %. The obtained matrix effects and accuracy values were also within acceptable ranges. Compounds 2 (effusol) and 3 (dehydroeffusol) were present in both methanolic and dichloromethanolic extracts of Plants 1 and 3 at the highest concentrations. Furthermore, the relationship between phenanthrene fingerprints, obtained through ANOVA statistical analysis of quantitative data, and the geographical location of herbs was also established.

Impact of size and fragmentation of the anteroinferior glenoid rim on clinical and functional outcomes of non-operatively treated Bony Bankart lesions in middle-aged population.

INTRODUCTION: The optimal treatment approach for Bony Bankart remains a subject of considerable debate among shoulder surgeons. Existing literature highlights low recurrence rates and high patient satisfaction with nonoperative treatment, particularly in the middle-aged population. This study aimed to evaluate the recurrence rate of dislocation, as well as the clinical and functional outcomes in middle-aged individuals treated nonoperatively following an acute bony Bankart fracture. Additionally, the impact of glenoid rim size and fragmentation on the treatment outcome was investigated. MATERIAL AND METHODS: A prospective analysis was conducted on 20 patients aged over 50 with nonoperatively treated bony Bankart fractures, ensuring a minimum follow-up of 24 months. The study population was categorized based on fragment size (small and medium) according to Kim classification and glenoid rim fragmentation (type 1b and 1c) according to Scheibel classification. Data including UCLA score, Rowe score, recurrence rate, clinical instability, and range of motion (ROM) were collected and analyzed. RESULTS: The average UCLA and Rowe scores were 32.15 ± 2.85 and 93.85 ± 2.19, respectively, with no instances of dislocation recurrence. The affected shoulder exhibited no significant reductions in ROM compared to the contralateral side, except for a loss of external rotation (ER) (13.08° ± 7.51; p = 0.005). No differences were observed based on fragment size, although patients with multifragmented glenoid rims showed a greater loss of ER compared to those with a solitary fragment, albeit not reaching statistical significance. CONCLUSION: Nonoperative treatment appears to be a viable and effective option for middle-aged individuals with bony Bankart fractures, resulting in favorable functional outcomes and a low risk of recurrence. Additionally, a notable loss of external rotation was observed in fractures with glenoid rim fragmentation. LEVEL OF EVIDENCE: IV.

Fragmentation of care in breast cancer: greater than the sum of its parts.

INTRODUCTION: Fragmentation of care (FC, the receipt of care at > 1 institution) has been shown to negatively impact cancer outcomes. Given the multimodal nature of breast cancer treatment, we sought to identify factors associated with FC and its effects on survival of breast cancer patients. METHODS: A retrospective analysis was performed of surgically treated, stage I-III breast cancer patients in the 2004-2020 National Cancer Database, excluding neoadjuvant therapy recipients. Patients were stratified into two groups: FC or non-FC care. Treatment delay was defined as definitive surgery > 60 days after diagnosis. Multivariable logistic regression was performed to identify factors predictive of FC, and survival was compared using Kaplan-Meier and multivariable Cox proportional hazards methods. RESULTS: Of the 531,644 patients identified, 340,297 (64.0%) received FC. After adjustment, FC (OR 1.27, 95% CI 1.25-1.29) was independently associated with treatment delay. Factors predictive of FC included Hispanic ethnicity (OR 1.04, 95% CI: 1.01-1.07), treatment at comprehensive community cancer programs (OR 1.06, 95% CI: 1.03-1.08) and integrated network cancer programs (OR 1.55, 95% CI: 1.51-1.59), AJCC stage II (OR 1.06, 95% CI 1.05-1.07) and stage III tumors (OR 1.06, 95% CI: 1.02-1.10), and HR + /HER2 + tumors (OR 1.05, 95% CI: 1.02-1.07). Treatment delay was independently associated with increased risk of mortality (HR 1.23, 95% CI 1.20-1.26), whereas FC (HR 0.87, 95% CI 0.86-0.88) showed survival benefit. CONCLUSIONS: While treatment delay negatively impacts survival in breast cancer patients, our findings suggest FC could be a marker for multispecialty care that may mitigate some of these effects.

UPLC-Q-TOF-MS/MS analysis of ophiobolins sesterterpenoids and bioactive analogs from Bipolaris eleusines.

In order to elucidate the mass fragmentation patterns and unveil more undescribed ophiobolin analogs, the mass fragmentation patterns of ophiobolins were analyzed based on UPLC-Q-TOF-MS/MS experiments. Different kinds of rearrangements (including McLafferty rearrangement) were the main cleavage patterns. Twenty-six (9-31) analogs were then tentatively characterized based on their mass analysis, and three undescribed ophiobolins (6-8) and a known analogue (5) were isolated in target. Compound 5 possesses a rare polycyclic carbon skeleton only recently reported, and compound 6 contains an undescribed lactone ring system fused with A/B ring at C-3/C-21, whereas compounds 7 and 8 have a peroxyl group in the side chain, which is the first reported in all ophiobolins. Compounds 5 and 7 displayed significant cytotoxicity against MCF-7 cancer cells.

Bottom-up Histone Post-translational Modification Analysis using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Tandem Mass Spectrometry.

The amino acid position within a histone sequence and the chemical nature of post-translational modifications (PTMs) are essential for elucidating the "Histone Code". Previous work has shown that PTMs induce specific biological responses and are good candidates as biomarkers for diagnostics. Here, we evaluate the analytical advantages of trapped ion mobility (TIMS) with parallel accumulation-serial fragmentation (PASEF) and tandem mass spectrometry (MS/MS) for bottom-up proteomics of model cancer cells. The study also considered the use of nanoliquid chromatography (LC) and traditional methods: LC-TIMS-PASEF-ToF MS/MS vs nLC-TIMS-PASEF-ToF MS/MS vs nLC-MS/MS. The addition of TIMS and PASEF-MS/MS increased the number of detected peptides due to the added separation dimension. All three methods showed high reproducibility and low RSD in the MS domain (<5 ppm). While the LC, nLC and TIMS separations showed small RSD across samples, the accurate mobility (1/K0) measurements (<0.6% RSD) increased the confidence of peptide assignments. Trends were observed in the retention time and mobility concerning the number and type of PTMs (e.g., ac, me1-3) and their corresponding unmodified, propionylated peptide that aided in peptide assignment. Mobility separation permitted the annotation of coeluting structural and positional isomers and compared with nLC-MS/MS showed several advantages due to reduced chemical noise.

Segmentation of beach plastic fragments' contours based on self-organizing map and multi-shape descriptors: A rapid indication of fragmentation and wearing types.

Environmental plastic fragments have been verified as byproducts of large plastic and its secondary pollutants including micro and nanoplastics. There are few quantitative studies available, but their contours have values for the weathering mechanisms. We used geometric descriptors, fractal dimensions, and Fourier descriptors to characterize field and artificial polyethylene and polypropylene samples as a means of investigating the contour characteristics. It provides a methodological framework for contour classification. Unsupervised classification was performed using self-organizing neural networks with size-invariance parameters. We revealed the isometric phenomenon of plastic fragments during fragmentation, i.e., that the degree of contour rounding and complexity increase and decrease, respectively, with decreasing fragment size. With an average error rate of 8.9 %, we can distinguish artificial samples from field samples. It was also validated by the difference in Carbonyl Index between groups. We propose a two-stage process for plastic fragmentation and give three types of contour features which were key in the description of fragmented contours, i.e., size, complexity, and rounding. Our work will improve the accuracy of characterizations regarding the weathering and fragmentation processes of certain kinds of plastic fragments. The contour parameters also have the potential to be applied in more realistic scenarios and varied polymers.

Esterified styrene-maleic acid copolymer modified silica as mixed-mode polymer-brush stationary phases for chromatographic separation.

Styrene-maleic acid (SMA) copolymer has received much attention for its excellent solubilization characteristics. In this work, SMA copolymer brush-based chromatographic stationary phases were exploited and developed for the first time. First, SMA copolymer brush was in situ grown on the surface of spherical silica via living/controlled reversible addition-fragmentation chain transfer (RAFT) polymerization method. Subsequently, as a proof-of-concept demonstration, the copolymer was esterified by diethylene glycol mono-2-ethylhexyl ether (DGME) and 2-(2-ethylhexyloxy) ethanol (EHOE), respectively. The obtained Sil-SMA-DGME and Sil-SMA-EHOE copolymer-brush chromatographic stationary phases were characterized by transmission electron microscopy, Fourier transform infrared spectrometer, X-ray photoelectron spectroscopy, and thermogravimetric analysis, respectively. The chromatographic retention mechanism indicated that both the two packed columns exhibited hydrophilic/reverse mixed-mode retention modes. The maximum column efficiency was up to 71,000 N/m. The chromatographic separation performance evaluation indicated that the novel kind of stationary phases had excellent separation capabilities for hydrophilic, hydrophobic compounds and phospholipid standards. In addition, by combination with mass spectrometry identification, the Sil-SMA-DGME column was further exploited for separation and identification of phospholipids in human lung cancer cells. Totally, 9 classes including 186 phospholipid species were successfully identified. The results demonstrated the promising application prospects of the novel kind of SMA copolymer-brush chromatographic stationary phases.

A comprehensive gas chromatography electron ionization high resolution mass spectrometry study of a new steroidal selective androgen receptor modulator (SARM) compound S42.

The synthetic 20-keto-steroid S42 (1) demonstrated selective androgen receptor modulator (SARM) properties in preclinical studies and, consequently, received growing attention also in the context of sports drug testing programs. Fundamental understanding of the behavior of S42 (1) and of relevant derivatives in gas chromatography-electron ionization MS experiments at high resolution (GC-EI-HRMS) is indispensable to develop a reliable qualitative and quantitative doping control method for S42 (1) and its metabolites in body fluid matrices. We present important fundamental mechanistic data on the EI fragmentation behavior of S42 (1) and of silyl ether derivatives as well as of stable isotope-labelled reference material.

Lens Fragmentation with Picosecond Laser Pulses After Artificial Cataract Induction with Microwaves.

Objectives: In this work we demonstrate the first laboratory study results of lens fragmentation with low-energy picosecond ultrashort laser pulses after artificial induction of cataract with microwave radiation on an ex vivo animal model. Background: This method will be evaluated with regard to the further development of lens fragmentation with novel ultrashort picosecond laser systems instead of ultrasonic phacoemulsification or the significantly more complex femtosecond laser fragmentation. Methods: As samples we used postmortem porcine eyes. The lenses were dissected and then irradiated in a microwave oven for artificial cataract induction. Subsequent computer-driven lens fragmentation was performed with a 12 ps, 1064 nm pulsed laser source with 100 µJ pulse energy, and 10 kHz pulse repetition rate. Results: Both the artificial cataract induction and the lens fragmentation were demonstrated. When inducing cataract, different degrees/stages of opaqueness and hardness could be achieved with different irradiation times and methods. The fragmentation with 12 ps pulses led to good results with regard to ablation depth and rate, especially for the softer lenses. Conclusions: As could be shown, low-energy picosecond ultrashort laser pulses are feasible for cataractous lens fragmentation on an ex vivo animal model with artificial cataract induction. Thus, this technique may influence future cataract surgeries by possibly being an alternative or extension to state-of-the-art methods. This will be evaluated with further tests and studies.

Pilot screening of potential matrikines resulting from collagen breakages through ionizing radiation.

Little is known regarding radiation-induced matrikines and the possible degradation of extracellular matrix following therapeutic irradiation. The goal of this study was to determine if irradiation can cut collagen proteins at specific sites, inducing potentially biologically active peptides against cartilage cells. Chondrocytes cultured as 3D models were evaluated for extracellular matrix production. Bystander molecules were analyzed in vitro in the conditioned medium of X-irradiated chondrocytes. Preferential breakage sites were analyzed in collagen polypeptide by mass spectrometry and resulting peptides were tested against chondrocytes. 3D models of chondrocytes displayed a light extracellular matrix able to maintain the structure. Irradiated and bystander chondrocytes showed a surprising radiation sensitivity at low doses, characteristic of the presence of bystander factors, particularly following 0.1 Gy. The glycine-proline peptidic bond was observed as a preferential cleavage site and a possible weakness of the collagen polypeptide after irradiation. From the 46 collagen peptides analyzed against chondrocytes culture, 20 peptides induced a reduction of viability and 5 peptides induced an increase of viability at the highest concentration between 0.1 and 1 µg/ml. We conclude that irradiation promoted a site-specific degradation of collagen. The potentially resulting peptides induce negative or positive regulations of chondrocyte growth. Taken together, these results suggest that ionizing radiation causes a degradation of cartilage proteins, leading to a functional unbalance of cartilage homeostasis after exposure, contributing to cartilage dysfunction.

Determining KRAS4B-Targeting Compound Specificity by Top-Down Mass Spectrometry.

We present a novel method to determine engagement and specificity of KRAS4B-targeting compounds in vitro. By employing top-down mass spectrometry (MS), which analyzes intact and modified protein molecules (proteoforms), we can directly visualize and confidently characterize each KRAS4B species within compound-treated samples. Moreover, by employing targeted MS2 fragmentation, we can precisely localize each compound molecule to a specific residue on a given KRAS4B proteoform. This method allows us to comprehensively evaluate compound specificity, clearly detect nonspecific binding events, and determine the order and frequency with which they occur. We provide two proof-of-concept examples of our method employing publicly available compounds, along with detailed protocols for sample preparation, top-down MS data acquisition, targeted proteoform MS2 fragmentation, and analysis of the resulting data. Our results demonstrate the concentration dependence of KRAS4B-compound engagement and highlight the ability of top-down MS to directly map compound binding location(s) without disrupting the KRAS4B primary structure. Our hope is that this novel method may help accelerate the identification of new successful targeted inhibitors for KRAS4B and other RAS isoforms.

A novel polyp retrieval bag reduces the polyp fragmentation rate in colon polypectomy: a single-blind randomized controlled study.

PURPOSE: The fragmentation of polyps affects complete resection confirmation. The primary aim of this study was to assess the feasibility of a novel polyp retrieval bag for reducing the fragmentation rate of colon polyps. METHODS: Patients with a 5-15 mm colon polyp were recruited and randomized into two groups at a 1:1 ratio. After polyp resection, polyps obtained from patients in the treatment group were extracted via a novel polyp retrieval bag without traversing the instrument channel, whereas polyps obtained from patients in the control group were collected through the instrument channel, attaching the polyp trap to the instrument channel port, and applying suction. RESULTS: From January to July 2022, 225 patients were assessed for eligibility. The study participants included 204 patients, and seven patients whose samples were not retrieved were excluded. Polyp fragmentation was significantly lower in the treatment group than in the control group (3.0% [3/100] vs. 17.5% [17/97], P = 0.001). The retrieval failure rates in the treatment group and control group were not significantly different (2.0% [2/102] vs. 4.9% [5/102], P = 0.442). There were fewer colonoscope insertions in the treatment group than in the control group (102 vs. 110), but a significant difference was not present (P = 0.065). No significant adverse events were observed during the follow-up. CONCLUSIONS: This study demonstrated that the polyp retrieval bag was safe and feasible for reducing the fragmentation rate of retrieved polyps. TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov (NCT05189912, 1/12/2021).

The effects of Wharton's jelly MSCs secretomes for restoring busulfan-induced reproductive toxicity in male mice.

Several studies investigated the application of Mesenchymal stem cells (MSCs) for treating spermatogenic disorders. Considering the limitation of MSC application, the present study aimed to compare Wharton's jelly MSCs secretomes, including condition medium (CM) 10-fold concentrated (CM10), 20-fold concentrated CM (CM20), and extracellular vesicles (EVs) to restore busulfan-induced damage on male mice reproduction. So, Wharton's jelly MSCs were cultured, CM was collected, and EVs were isolated. Seventy-two mice were randomly assigned to nine groups, including Control, Busulfan 1 month (1M), Busulfan 2 months (2M), CM10, Busulfan + CM10, CM20, Busulfan + CM20, EVs, and Busulfan + EVs groups. Sperm characteristics, DNA maturity, DNA fragmentation index (DFI), and testicular gene expression were evaluated. Data analysis revealed that CM10 significantly improved sperm plasma membrane integrity, sperm DNA maturity, and DFI in the Busulfan + CM10 group compared to the Busulfan 2M group. Although CM20 and EVs showed a non-significant improvement. Gene expression analysis showed busulfan administration significantly decreased the expression of AR, CREB1, and PLCζ genes, while CM10 significantly restored CREB1 gene expression. The present study demonstrated that CM10 is more effective than CM20 or EVs in reducing busulfan-induced reproductive toxicity.

Preparation, characterization and antioxidant and anticancerous potential of Quercetin loaded ß-glucan particles derived from mushroom and yeast.

ß-glucans are polysaccharides found in the cell walls of various fungi, bacteria and cereals. ß-glucan have been found to show various kinds of anti-inflammatory, antimicrobial, antidiabetic antioxidant and anticancerous activities. In the present study, we have isolated ß-glucan from the baker's yeast Saccharomyces cerevisiae and white button mushroom Agaricus bisporus and tested their antioxidant potential and anticancerous activity against prostate cancer cell line PC3. Particles were characterized with zeta sizer and further with FTIR that confirmed that the isolated particles are ß-glucan and alginate sealing made slow and sustained release of the Quercetin from the ß-glucan particles. Morphological analysis of the hollow and Quercetin loaded ß-glucan was performed with the SEM analysis and stability was analyzed with TGA and DSC analysis that showed the higher stability of the alginate sealed particles. Assessments of the antioxidant potential showed that Quercetin loaded particles were having higher antioxidant activity than hollow ß-glucan particles. Cell viability of the PC3 cells was examined with MTT assay and it was found that Quercetin loaded alginate sealed Agaricus bisporus derived ß-glucan particles were having lowest IC50. Further ROS generation was found to increase in a dose dependent manner. Apoptosis detection was carried out with Propidium iodide and AO/EtBr staining dye which showed significant death in the cells treated with higher concentration of the particles. Study showed that particles derived from both of the sources were having efficient anticancer activity and showing a dose dependent increase in cell death in PC3 cells upon treatment.

DNA fragmentation of lymphocytes and sperm cells induced by nickel released from orthodontic archwires: A preliminary study.

Orthodontic brackets and archwires placed intraorally are subject to corrosion, leading to the release of cytotoxic metal ions. The aim of this study was to determine whether the use of orthodontic NiTi archwires increases systemic Ni levels and cause alterations on the DNA of cells unrelated to the oral environment such as lymphocytes and sperm cells. Human urine, semen and blood samples were collected before (baseline) sham placement of orthodontic archwires and 15 and 30 days after placement. Lymphocytes and sperm cells cells were evaluated by comet assay. Ni concentration levels in urine increased significantly between baseline and 15 days (p<0.01) and 15 and 30 days of exposure (p<0.01). Progressive decrease in sperm viability and motility was observed between the sampling periods. Lymphocytes and sperm cells showed DNA fragmentation. The increase in systemic concentration of nickel induced structural damage in the DNA of lymphocytes and human sperm cells.

Lead and calcium crosstalk tempted acrosome damage and hyperpolarization of spermatozoa: signaling and ultra-structural evidences.

BACKGROUND: Exposure of humans and animals to heavy metals is increasing day-by-day; thus, lead even today remains of significant public health concern. According to CDC, blood lead reference value (BLRV) ranges from 3.5 µg/dl to 5 µg/dl in adults. Recently, almost 2.6% decline in male fertility per year has been reported but the cause is not well established. Lead (Pb2+) affects the size of testis, semen quality, and secretory functions of prostate. But the molecular mechanism(s) of lead toxicity in sperm cells is not clear. Thus, present study was undertaken to evaluate the adverse effects of lead acetate at environmentally relevant exposure levels (0.5, 5, 10 and 20 ppm) on functional and molecular dynamics of spermatozoa of bucks following in vitro exposure for 15 min and 3 h. RESULTS: Lead significantly decreased motility, viable count, and motion kinematic patterns of spermatozoa like curvilinear velocity, straight-line velocity, average path velocity, beat cross frequency and maximum amplitude of head lateral displacement even at 5 ppm concentration. Pb2+ modulated intracellular cAMP and Ca2+ levels in sperm cells through L-type calcium channels and induced spontaneous or premature acrosome reaction (AR) by increasing tyrosine phosphorylation of sperm proteins and downregulated mitochondrial transmembrane potential. Lead significantly increased DNA damage and apoptosis as well. Electron microscopy studies revealed Pb2+ -induced deleterious effects on plasma membrane of head and acrosome including collapsed cristae in mitochondria. CONCLUSIONS: Pb2+ not only mimics Ca2+ but also affects cellular targets involved in generation of cAMP, mitochondrial transmembrane potential, and ionic exchange. Lead seems to interact with Ca2+ channels because of charge similarity and probably enters the sperm cell through these channels and results in hyperpolarization. Our findings also indicate lead-induced TP and intracellular Ca2+ release in spermatozoa which in turn may be responsible for premature acrosome exocytosis which is essential feature of capacitation for fertilization. Thus, lead seems to reduce the fertilizing capacity of spermatozoa even at 0.5 ppm concentrations.

Effect of glucocorticoid blockade on inflammatory responses to acute sleep fragmentation in male mice.

The association between sleep and the immune-endocrine system is well recognized, but the nature of that relationship is not well understood. Sleep fragmentation induces a pro-inflammatory response in peripheral tissues and brain, but it also activates the hypothalamic-pituitary-adrenal (HPA) axis, releasing glucocorticoids (GCs) (cortisol in humans and corticosterone in mice). It is unclear whether this rapid release of glucocorticoids acts to potentiate or dampen the inflammatory response in the short term. The purpose of this study was to determine whether blocking or suppressing glucocorticoid activity will affect the inflammatory response from acute sleep fragmentation (ASF). Male C57BL/6J mice were injected i.p. with either 0.9% NaCl (vehicle 1), metyrapone (a glucocorticoid synthesis inhibitor, dissolved in vehicle 1), 2% ethanol in polyethylene glycol (vehicle 2), or mifepristone (a glucocorticoid receptor antagonist, dissolved in vehicle 2) 10 min before the start of ASF or no sleep fragmentation (NSF). After 24 h, samples were collected from brain (prefrontal cortex, hypothalamus, hippocampus) and periphery (liver, spleen, heart, and epididymal white adipose tissue (EWAT)). Proinflammatory gene expression (TNF-α and IL-1ß) was measured, followed by gene expression analysis. Metyrapone treatment affected pro-inflammatory cytokine gene expression during ASF in some peripheral tissues, but not in the brain. More specifically, metyrapone treatment suppressed IL-1ß expression in EWAT during ASF, which implies a pro-inflammatory effect of GCs. However, in cardiac tissue, metyrapone treatment increased TNF-α expression in ASF mice, suggesting an anti-inflammatory effect of GCs. Mifepristone treatment yielded more significant results than metyrapone, reducing TNF-α expression in liver (only NSF mice) and cardiac tissue during ASF, indicating a pro-inflammatory role. Conversely, in the spleen of ASF-mice, mifepristone increased pro-inflammatory cytokines (TNF-α and IL-1ß), demonstrating an anti-inflammatory role. Furthermore, irrespective of sleep fragmentation, mifepristone increased pro-inflammatory cytokine gene expression in heart (IL-1ß), pre-frontal cortex (IL-1ß), and hypothalamus (IL-1ß). The results provide mixed evidence for pro- and anti-inflammatory functions of corticosterone to regulate inflammatory responses to acute sleep loss.

Organization of the Nuclei in the Tegument of Some Palaeacanthocephala and Archiacanthocephala.

The giant tegument nuclei of the acanthocephalans of the classes Archiacanthocephala and Palaeacanthocephala are fragmented at the final stage of cystacanthus formation in the intermediate host, but remain connected with each other during later life. It can be assumed that the fragments of each giant tegument nucleus are united with each other to form an independent network that ensures the vital activity of the tegument, the volume of which increases many times during the period of intensive growth of the parasite in the definitive host.