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The underlying pathophysiological mechanisms of hepatic ischemia-reperfusion (I/R) injury have not been completely elucidated. However, it is well known that oxidative stress, caused by a burst of reactive oxygen species (ROS) production during the reperfusion phase, plays a crucial role. A growing body of evidence indicates that the intracellular availability of free iron represents a requirement for ROS-induced adverse effects, as iron catalyzes the generation of highly reactive free radicals. The aim of this study was to examine whether a combination of iron chelators with varying lipophilicity could offer enhanced protection against I/R by diminishing the conversion of weak oxidants, like HHepG2 cells (hepatocellular carcinoma cell line) were exposed to oxidative stress conditions after pre-treatment with the iron chelators desferrioxamine (DFO) and deferiprone (DFP) alone or in combination. Labile iron pool was estimated using the calcein-acetoxymethyl ester (calcein-AM) method and DNA damage with the comet assay. We subsequently used a rabbit model (male New Zealand white rabbits) of hepatic I/R-induced injury to investigate, by measuring biochemical (ALT, ALT, ALP, γGT) and histological parameters, whether this may be true for in vivo conditions.The combination of a membrane-permeable iron chelator (DFP) with a strong membrane-impermeable one (DFO) raises the level of protection in both hepatic cell lines exposed to oxidative stress conditions and hepatic I/R rabbit model.Our results show that combinations of iron chelators with selected lipophilicity and iron-binding properties may represent a valuable strategy to protect against tissue damage during reperfusion after a period of ischemia.

In December 2019, the new coronavirus infection (COVID-19) was declared a pandemic by the World Health Organization after rapidly spreading over the world in just a few months. All elective operations and nonemergency treatments have been postponed worldwide. However, some patients require surgical therapy as well, and the time spent waiting should not have a negative impact on the surgical outcome or disease course. Following the initial onset of the COVID-19 epidemic, instructions for proper and safe surgery for healthcare staff and patients should develop. Thyroid surgeries have decreased during the COVID-19 pandemic. Most of them can be postponed for a long time. Assessment of thyroid nodules recommends clinical examination, imaging studies, fine needle aspiration (FNA) and vocal cord examination. All these procedures are necessary, and sometimes they cannot be postponed. To determine the best timing, a thorough preoperative assessment should be undertaken, taking into account both oncological and anatomical features. Furthermore, COVID-19 status must be negative prior to any intervention, and hospital infrastructure must be ready to deal with the demanding situation.

Objectives Surgery of the thyroid takes place in a body part with complicated anatomy and several vital physiologic functions. Thyroidectomy is rarely associated with mortality but can be followed by significant complications, (i.e. hypoparathyroidism, hemorrhage, upper airway obstruction, laryngeal nerve injuries and thyrotoxic storm). This review aims to indicate surgical tips and techniques to sustain a low level of complications. Content MEDLINE database (PubMed) platform was used as a search engine and the articles related to the topic were selected using the keywords combination “thyroid surgery and complications”. Summary and Outlook The most common complication of total thyroidectomy with an occurrence ranging between 0.5 and 65% is hypoparathyroidism. Damage to recurrent laryngeal nerves can be temporary or permanent, unilateral or bilateral; bilateral lesion is associated with severe episodes of breathlessness. Thus, intraoperative monitoring of nerve function is essential to prevent damage. Ιn addition, hematoma formation can lead to breathing difficulties due to airway obstruction; preventive hemostasis during surgery is essential. The surgeon must have a complete anatomical understanding of not only the normal anatomy of the central visceral compartment of the neck, but also the common variations of the laryngeal nerves and parathyroid glands in order to keep the complication rate at a very low level.

Objectives The great saphenous vein is the lower limb’s longest and thickest-walled superficial vein. Its anatomy is complex, while its anatomical variations are widespread. This study aimed to illustrate the anatomy and variations of the great saphenous vein at the saphenofemoral junction. Methods The study was conducted on 75 fresh-frozen cadaveric lower limbs. Cadavers with evidence of prior leg surgery or trauma and any congenital abnormality or gross deformity were excluded. The saphenofemoral junction was studied in detail with emphasis on the number, incidence, draining pattern of tributaries, and duplication of the great saphenous vein. Results Out of the 75 cadaveric lower limbs, 57.3% were obtained from males, and 42.7% were obtained from females. The mean age of the cadaveric lower extremities was 66.6 years (range 42–91). The number of tributaries at the saphenofemoral junction varied from 0 to 7, with a mean of 3.8. The most frequent number of branches was 4–5 in 61.3% of cases. The most consistent tributary was the superficial external pudendal vein, while the posterior accessory great saphenous vein was the least frequent tributary. 80% of the branches drained into the saphenofemoral junction directly (53.3%) or by a common trunk (26.7%%). The most frequent common trunk was the superficial epigastric and superficial circumflex iliac vein (10.7%). The rest 20% of the tributaries drained directly into the common femoral vein. The commonest branch reaching the common femoral vein was the superficial external pudendal vein (10.7%). Duplication of the great saphenous vein was observed in 2.7% of the cases. Conclusions There is a significant variation of tributaries at the saphenofemoral junction regarding the number, incidence, draining pattern of branches, and duplication of the great saphenous vein.

Summary. Phlebosclerosis is a venous wall degenerative disease which has gained little popularity in the literature due to its uncertain clinical significance. The objective of this review is to evaluate the epidemiology, etiology and clinical significance of phlebosclerosis in lower extremities veins, particularly the effect of preexisting phlebosclerosis of the great saphenous vein on vein graft patency. Medline was searched from inception until November 1, 2019. Reference lists of included studies were scanned. Only articles published after 1949 were included. Two reviewers independently screened titles/abstracts and full-text papers for any study design in relation to phlebosclerosis in lower extremities veins and abstracted data. A total of 16 Cohort studies and one case-control study (3708 participants, mean age 61.8 years, 59.3 % men, and 40.7 % women) were included after screening 317 titles and abstracts, and 80 full-text articles. The incidence of phlebosclerosis ranged from 1.5–9.7 % depending on the radiological features. On the contrary, the incidence of the phlebosclerotic great saphenous vein prior to its use as a vein graft was 26.9–91 % on histological examination. The small saphenous vein was the most common location of phlebosclerosis followed by the great saphenous vein. There is a link between phlebosclerosis and age, venous insufficiency and haemodialysis. As for the vein graft patency seven studies demonstrated a correlation between preexisting phlebosclerosis and vein graft stenosis, whereas three studies failed to prove any association. In conclusion, the radiological incidence of phlebosclerosis depended on the ultrasound findings. Its presence in the great saphenous vein prior to its use as a vein graft is established on histological examination. The small saphenous vein is mainly affected. Risk factors included age, haemodialysis, and venous insufficiency. Preexisting wall thickness of the great saphenous vein graft seemed to affect negatively its patency in bypass surgery.

Adamantinoma is a biphasic tumor, with a low potential for malignancy, characterized by clusters of epithelial cells surrounded by a relatively bland spindle-cell osteofibrous component. The aim of the present study was to review the updated data regarding epidemiology; pathogenesis; clinical presentation; radiological, histopathological and ultrastructural findings; and treatment options of adamantinoma. In X-ray, it is usually seen as an eccentric and sometimes central, lobular, lytic lesion with sclerotic margins of overlapping radiolucency, and a characteristic 'soap-bubble' appearance. Magnetic resonance imaging seems to be the most appropriate examination for differential diagnosis between adamantinoma and other skeletal tumors. Histologically, adamantinoma is identified as classic adamantinoma or osteofibrous-like adamantinoma. Classic adamantinoma is classified into four patterns of growth: Basaloid, tubular, spindle cell, and squamous. The preferable treatment of this tumor type is en bloc resection within wide operative margins, which may include suspicious regional lymph nodes, with limb reconstruction and limb salvage.

Wine and by-products are essential elements of a Mediterranean diet and considered as a reservoir of bioactive compounds with various health effects. Grape pomace, an easily available natural material of low cost, shares a similar wealth of health benefiting bioactive phytochemicals. The objective of this study was to explore the utilization of grape pomace from Commandaria dessert wine as main ingredient for functional infusions. Therefore, the ratio of water to grape pomace powder (40–200 mL g−1), infusion time (3–15 min) and temperature (55–95 °C) were optimized in terms of composition and bioactivity. Multiple response optimization indicated that brewing 200 mL water per g of material for 12.2 min at 95 °C, was the optimum method for preparing the infusion. Results also revealed a significant impact of three parameters as well as quadratic and interactive effects on composition and bioactivity of infusions. Furthermore, the infusion presents antimicrobial effects against Listeria monocytogenes serotypes and other common food pathogenic bacteria. Finally, a sensory evaluation was performed to assess the organoleptic attributes of the infusion and its improvement, with the addition of Mediterranean aromatic plants. Overall, the present work describes a promising strategy for the re-use of sun-dried grape pomace as a functional ingredient of infusions.

Osteoid osteoma, the third most common benign bone tumor, usually occurs in the cortex of long bones. It consists of a radiolucent nidus surrounded by reactive osteosclerosis. Generally, osteoid osteoma affects young males. Nocturnal pain that eases with salicylates or nonsteroidal anti-inflammatory drugs (NSAID) is the typical clinical presentation. Sometimes, it remains undiagnosed for a long time. Plain radiography and computed tomography are usually sufficient for the diagnosis of osteoid osteoma. Initial treatment includes salicylates and NSAID because the tumor often regresses spontaneously over 2-6 years. Surgical treatment is indicated in case of unresponsive pain to medical therapy, no tolerance of prolonged NSAID therapy due to side effects, and no willingness to activity limitations. Nowadays, minimally invasive techniques have replaced open surgery and are considered the gold standard of surgical treatment. Although cryoablation seems superior in terms of the nerve damage and immunotherapy effect, radiofrequency ablation is the preferred technique.

Osteoid osteoma, the third most common benign bone tumor, usually occurs in the cortex of long bones. It consists of a radiolucent nidus surrounded by reactive osteosclerosis. Generally, osteoid osteoma affects young males. Nocturnal pain that eases with salicylates or nonsteroidal anti-inflammatory drugs (NSAID) is the typical clinical presentation. Sometimes, it remains undiagnosed for a long time. Plain radiography and computed tomography are usually sufficient for the diagnosis of osteoid osteoma. Initial treatment includes salicylates and NSAID because the tumor often regresses spontaneously over 2-6 years. Surgical treatment is indicated in case of unresponsive pain to medical therapy, no tolerance of prolonged NSAID therapy due to side effects, and no willingness to activity limitations. Nowadays, minimally invasive techniques have replaced open surgery and are considered the gold standard of surgical treatment. Although cryoablation seems superior in terms of the nerve damage and immunotherapy effect, radiofrequency ablation is the preferred technique.

One of the prevailing perceptions regarding the ageing of cells and organisms is the intracellular gradual accumulation of oxidatively damaged macromolecules, leading to the decline of cell and organ function (free radical theory of ageing). This chemically undefined material known as “lipofuscin,” “ceroid,” or “age pigment” is mainly formed through unregulated and nonspecific oxidative modifications of cellular macromolecules that are induced by highly reactive free radicals. A necessary precondition for reactive free radical generation and lipofuscin formation is the intracellular availability of ferrous iron (Fe2+) (“labile iron”), catalyzing the conversion of weak oxidants such as peroxides, to extremely reactive ones like hydroxyl (HO•) or alcoxyl (RO•) radicals. If the oxidized materials remain unrepaired for extended periods of time, they can be further oxidized to generate ultimate over-oxidized products that are unable to be repaired, degraded, or exocytosed by the relevant cellular systems. Additionally, over-oxidized materials might inactivate cellular protection and repair mechanisms, thus allowing for futile cycles of increasingly rapid lipofuscin accumulation. In this review paper, we present evidence that the modulation of the labile iron pool distribution by nutritional or pharmacological means represents a hitherto unappreciated target for hampering lipofuscin accumulation and cellular ageing.

Apart from triglycerides, olive oil contains low quantities of polar compounds, such as polyphenols, which are believed to be relevant for maintaining human health. In this presentation, we will discuss potential molecular mechanisms by which specific phenolic compounds present in olive oil can exert cytoprotective and cell signal modulating effects, through chelation of intracellular labile (redox-active) iron. Iron chelation by specific polyphenols can prevent or relocalize the interaction of iron with hydroperoxides produced in the cells and the formation of extremely reactive species. The latter are able, either to induce uncontrolled oxidation of basic cell components or to modulate redox signaling. It is proposed that olive oil–contained polyphenols can influence the outcome of cell responses in conditions of increased oxidative stress mainly by this way.

The human thymus is a primary lymphoepithelial organ which supports the production of self-tolerant T cells with competent and regulatory functions. Paradoxically, despite the crucial role that it exerts in T cell-mediated immunity and prevention of systemic autoimmunity, the thymus is the first organ of the body that exhibits age-associated degeneration/regression, termed “thymic involution.” A hallmark of this early phenomenon is a progressive decline of thymic mass as well as a decreased output of naïve T cells, thus resulting in impaired immune response. Importantly, thymic involution has been recently linked with cellular senescence which is a stress response induced by various stimuli. Accumulation of senescent cells in tissues has been implicated in aging and a plethora of age-related diseases. In addition, several lines of evidence indicate that oxidative stress, a well-established trigger of senescence, is also involved in thymic involution, thus highlighting a possible interplay between oxidative stress, senescence, and thymic involution.

Cellular senescence, an age-related process in response to damage and stress, also occurs during normal development and adult life. The thymus is a central lymphoepithelial organ of the immune system that exhibits age-related changes termed thymic involution. Since the mechanisms regulating thymic involution are still not well elucidated, we questioned whether cellular senescence is implicated in this process. We demonstrate, for the first time in situ, that cellular senescence occurs during human thymic involution using SenTraGor™, a novel chemical compound that is applicable in archival tissue material, providing thus further insights in thymus histophysiology.

The purpose of this cadaveric study is to determine safe zones utilizing volar portals for wrist arthroscopy, by quantitatively describing the neurovascular relationships of a volar radial and a volar ulnar wrist arthroscopy portals in comparison with those of a newly described volar central portal (7) , considering the advantages in visualization of volar portals for wrist arthroscopy over the standard dorsal (19) . The neurovascular structures and the tendons of nine frozen human cadaveric upper limbs were exposed, while the aforementioned volar portal sites were pointed out with pins. The horizontal distance between the portals and the closest neurovascular branch or tendon was measured with a digital caliper, followed by statistical analysis of the data. The median interquartile range distances from portals to structures at risk were measured and safe zones around each portal were established. This study provides a safe approach to the volar radial and ulnar aspects of the radiocarpal and midcarpal joints, while volar radial and ulnar portals should be considered for inclusion in the arthroscopic examination of any patient with radial and ulnar sided wrist pain respectively (17,18) . Regarding the volar central portal, it is reproducible, safe and both the above joints can be inspected through one single incision (7) .

Juvenile idiopathic arthritis is a heterogeneous group of conditions encompassing all forms of unknown origin arthritis before the age of 16 years that persist for more than 6 weeks. It is the most common rheumatic disease in young patients and causes severe disabilities, thus an early initiation of the appropriate treatment modalities is necessary. First therapeutic options are nonsteroidal anti-inflammatory drugs, corticosteroids, and conventional non-biologic disease-modifying anti-rheumatic drugs, such as methotrexate. Insufficiency of these drugs led to the introduction of new biological medications that selectively target specific cytokines with an objective to suppress the disease. Despite the success in treatment and physical therapy, some of the patients develop advanced arthritis that can result in severe pain and disability. In such cases, surgical intervention is required to improve quality of life. The surgical methods include soft tissue release, osteotomies, synovectomies, and arthrodesis. Total joint replacement is the last option for endstage degenerative conditions (patients with deformity, poor motion, and severe pain). Deep infections, bone perforation, acetabular protrusion, postoperative dislocations, and the need for re-operation are some of the complications of total joint arthroplasty. This review summarizes published studies of the treatment of juvenile idiopathic arthritis focusing mainly on surgical treatment. Our purpose is to evaluate the general trends in treatment of juvenile idiopathic arthritis, focusing on methods, therapeutic advances, and outcomes of the intervention applied.

Introduction An awareness of the anatomical parameters of the foot such as the position and orientation of the sesamoid bones can be of great value for the etiology and diagnostic approach to patients with hallux valgus (HV). The purpose of this study was to evaluate the basic anatomical features and measurements related to the characteristics of HV in cadaveric material. Materials and methods The study sample included 12 cadaveric limbs with HV and 10 cadaveric limbs without HV as a control group. We measured the HV angle (HVA), the first to second intermetatarsal angle (IMA), and the first to fifth IMAs of all samples. We also recorded the shape of the first metatarsal head and the position/orientation of the sesamoid bones. Results The mean values of the HVA (p = 0.000), the first to second IMA (p = 0.000), and the first to fifth IMA (p = 0.000) differed between the HV and non-HV group. The position of the sesamoid ossicles between the HV and the non-HV group was statistically significant (p = 0.001). While we noted the round shape of the first metatarsal head was predominant in HV samples, we found no statistically significant difference in the first metatarsal head shape between the test and control groups. Conclusion The HVA, the first to second IMA, the first to fifth IMA, and the orientation of the sesamoid ossicles differed significantly between cadaveric samples with HV and those without HV.

The activator protein-1 (AP-1) is a dimeric transcription factor composed of proteins belonging to the Jun (c-Jun, JunB and JunD), Fos (c-Fos, FosB, Fra1 and Fra2) and activating transcription factor protein families. AP-1 is involved in various cellular events including differentiation, proliferation, survival and apoptosis. Deregulated expression of AP-1 transcription factors is implicated in the pathogenesis of various lymphomas such as classical Hodgkin lymphomas, anaplastic large cell lymphomas, diffuse large B cell lymphomas and adult T cell leukemia/lymphoma. The main purpose of this review is the analysis of the expression patterns of AP-1 transcription factors in order to gain insight into the histophysiology of lymphoid tissues and the pathology of lymphoid malignancies.

The objective of this study was to evaluate the contribution of the medial retinaculum to the stability of the patella, for several angles of knee flexion. For six cadaveric knees, the patella and patella retinaculum were exposed, and the force required to dislocate the patella laterally by 5 mm was measured, in 0, 45 and 90 degrees of knee flexion, preserving and dissecting the medial retinaculum. Wilcoxon singed rank test (SPSS, version 23, Chicago, USA) was used for data analysis. A p value of0.05 was considered as statistically significant. The force required to displace the patella laterally is smaller with the medial retinaculum dissected than intact, in 0, 45 and 90 degrees (p = 0.028, p = 0.046, p = 0.027 respectively). The lateral displacement force is greater as the flexion angle increases, with medial retinaculum intact or dissected (p = 0.028). Thus, an intact medial retinaculum provides more stability against lateral displacement forces to the patella, especially in lower flexion angles. Consequently, surgical methods reinforcing the medial retinaculum combined or not with lateral retinaculum release, are of great importance in the plan of the orthopedic surgeons. The engagement of the bones during flexion of the knee contributes significantly to the stability of the patella.

Long-standing scapholunate instability frequently leads to progressive deterioration of the adjacent joint cartilages in a consistent repetitive sequence of arthritis. The purpose of this study was to investigate whether the capitate-triquetrum distance is related with the occurrence of carpal collapse in cases of static scapholunate instability. In this retrospective study, 41 patients formed two groups based on the capitate-triquetrum distance; twenty-four with distance of less than 5mm and 17 with distance of 5mm or more. No significant difference was detected with respect to age, sex distribution, dominant hand involvement, initial treatment and time from injury to final x-rays. Three patients (3/24, 12.5%) of the C-T5mm group and 10 patients (10/17, 58.8%) of the C-T ≥ 5mm group had no sign of carpal collapse. This difference was statistically significant (p=0.017). Capitate-triquetrum distance could reliably contribute in the decision making process in difficult cases of static scapholunate instability.

Iron is a transition metal and essential constituent of almost all living cells and organisms. As component of various metalloproteins it is involved in critical biochemical processes such as transport of oxygen in tissues, electron transfer reactions during respiration in mitochondria, synthesis and repair of DNA, metabolism of xenobiotics, etc. However, when present in excess within cells and tissues, iron disrupts redox homeostasis and catalyzes the propagation of reactive oxygen species (ROS), leading to oxidative stress. ROS are critical for physiological signaling pathways, but oxidative stress is associated with tissue injury and disease. At the cellular level, oxidative stress may lead to ferroptosis, an iron-dependent form of cell death. In this review, we focus on the intimate relationship between iron metabolism and oxidative stress in health and disease. We discuss aspects of redox- and iron-mediated signaling, toxicity, ferroptotic cell death, homeostatic pathways and pathophysiological implications.

Background: The aim of this study was to analyse the biomechanical role of medial retinaculum, as a stabilising factor against lateral patellar dislocation. Materials and methods: This cadaveric-biomechanical study included the patellae of 10 cadaveric knees, which were surgically exposed and the medial retinaculum of each one was located. A stable 24.51 N force was applied to the four parts of the quadriceps, and an increasing lateral displacing force was applied to the patella, up to 5 mm dislocation. The study was repeated for 0o, 45o, and 90o of knee flexion, with the medial retinaculum intact and dissected. The Wilcoxon signed rank test was used for data analysis. A p value < 0.05 was considered as statistical significant. Results: After the dissection of medial retinaculum, the lateral displacement force was lower at every angle of knee flexion (p = 0.005, p = 0.007, p = 0.005, respectively). The lateral displacement force increased as the flexion angle increased (p = 0.005), regardless of medial retinaculum integrity. Conclusions: Medial retinaculum acts as a stabilising factor for the patella, against its lateral dislocation in lower flexion angles. Therefore, methods of surgical reinforcement or repair of medial retinaculum could provide protection against recurrent patellar dislocation.

Objective: To evaluate the correlation between the Manchester Grading Scale and the American Orthopaedic Foot and Ankle Society (AOFAS) score in patients with a hallux valgus deformity. Subjects and Methods: The study sample included 181 feet of 122 patients with hallux valgus and 424 feet of 212 individuals without hallux valgus deformity as the control group. The severity of hallux valgus, utilizing a relative nonmetric scale, the Manchester Grading Scale, and the metric AOFAS score, was determined for all individuals in the hallux valgus and control groups. SPSS version 18 (Chicago, Ill., USA) was used for data analysis. Results: According to the Manchester Grading Scale, the 424 feet of the normal group were classified as ‘no deformity'. In the hallux valgus group, 85 feet were classified as ‘mild deformity', 67 as ‘moderate deformity' and 29 as ‘severe deformity'. The AOFAS total score in the control group was 99.14. In the hallux valgus group, patients with mild or moderate deformity had total scores of 86.20 and 68.19, respectively. For those with severe hallux valgus, the total score was 44.69 and the differences were statistically significant (p = 0.000). Using the Pearson correlation, strong negative correlations were found between the AOFAS score and the hallux valgus angle (HVA; r = -0.899, p = 0.000). Strong negative correlations were demonstrated between the AOFAS score and the first intermetatarsal angle (IMA) as well (r = -0.748, p = 0.000). Conclusions: The AOFAS score was negatively associated with the Manchester Grading Scale, HVA and first IMA. As the severity of hallux valgus increased, the AOFAS score seemed to decrease.

Faculty of Science and Technology, Bournemouth University, Poole, UKSummaryOptimal conditions of short-term storage of fish sperm areimportant for successful artificial reproduction protocols orincubation of sperm in a cryoprotecting medium for success-ful sperm cryopreservation. The influence of acid or alkalineincubating conditions on short term storage of goldfishsperm was investigated in a series of experiments. Goldfishsperm was diluted an immobilizing solution (1 : 3) and incu-bated for 1 h in acidic, pH 6.5 (AC group) or alkaline, pH8.5 (AL group) conditions. Subsequently, the sperm of bothgroups was further diluted (1 : 3) in the immobilizing solu-tion and the pH was adjusted to pH 7.51 ( 0.02) and 7.56( 0.07) in the AC and AL group respectively. The sampleswere kept in sealed bags and stored at 4°C. Viability (%)and motility (%) was estimated in samples obtained daily forthe next 4 days. The results indicate that even a brief expo-sure of sperm to acid acidic conditions prior to storage accel-erated the decline of sperm viability and motility duringrefrigeration.IntroductionThere are several published protocols for chilled storage offish sperm (Mansour et al., 2004; Babiak et al., 2006;Pe~naranda et al., 2010a,b). A successful sperm preservationprotocol involves the dilution of semen in an immobilisingsolution, which protects the chemical and structural cellularintegrity of spermatozoa. An activation solution is requiredfor initiating the motility of the immobilised spermatozoa forthe fertilisation of egg. The integrity of spermatozoa requiresthe preservation of membrane integrity and the safeguardingof cellular functions and capacity for the reactivation andmotility of spermatozoa when kept in the immobilising solu-tion.Several motility parameters of sperm can be influenced bythe pH of the immobilising solution (Perchec et al., 1995;Woolsey and Ingermann, 2003) or by the ionic compositionand concentration, osmolarity, pH, and the dilution rate ofsperm in an activating solution (Alavi and Cosson, 2005).An immobilizing medium can control the ionic environmentand overcomes the problems of oxygenation and dehydrationduring chilled storage (Saad et al., 1988; Babiak et al.,2006).Sperm viability and motility are important parameters forevaluating the quality of sperm. There are several studiesthat demonstrate the negative effect of low pH on mamma-lian and fish sperm motility (Arienti et al., 1999; Nyncaet al., 2012). During sperm collection and handling, variousparameters can influence the pH of sperm. For example, thepH of sperm may vary with season, diet and the level ofcontamination with urine (Dreanno et al., 1998; Bozkurt andSecer, 2005). A drop in the pH can result in low motility,fertilization and hatching rate (Nynca et al., 2012). The exactinfluence of extracellular pH on sperm motility and viabilitymay vary with species and the acidic or alkaline range of thepH that the sperm is exposed (Dziewulska and Domagala,2013; Gonzalez-Bernat et al., 2013). Exposure of sperm toacidic conditions may result in lethal or sub-lethal cellulardamage and, in some cases, promote irreversible changes inthe cell physiology that affect the motility of the spermatozoa(Pe~naranda et al., 2010a,b; Santiso et al., 2012).The intracellular pH of spermatozoa can be influenced byexternal pH with consequences for sperm motility. The effectof external pH on sperm motility, during storage or afteractivation of spermatozoa, involves the regulation of mem-brane ion channels and regulation of intracellular pH andprotein posphorylation (Wang et al., 2003). As a result,extracellular acidic conditions can influence the intracellularpH, decrease sperm motility, the effect of which may be dueto depletion of energy stores and decreased ATP hydrolyticactivity of the outer arm of dynein (Perchec et al., 1995;Woolsey and Ingermann, 2003) with consequences for thecapacity of motility of the spermatozoa.In cyprinidae, sperm viability may not be affected in awide range of pH, but motility may be compromised byextreme pHs; for example, the motility of carp spermatozoaincubated for 24 h at pH 6 or 10 was significantly lowercompared to spermatozoa incubated at pH 7.8 or 9.0 (Saadet al., 1988). As a result, incubation of spermatozoa toextreme pH conditions may result in sublethal damages,which will result in compromised motility and even inreduced fertilisation ability. For example, the swimming

The fundamental role of human Toll-like receptors (TLRs) and NOD-like receptors (NLRs), the two most studied pathogen recognition receptors (PRRs), is the protection against pathogens and excessive tissue injury. Recent evidence supports the association between TLR / NLR gene mutations and susceptibility to inflammatory, autoimmune and malignant diseases. PRRs also interfere with several cellular processes, such as cell growth, apoptosis, cell proliferation, differentiation, autophagy, angiogenesis, cell motility and migration, and DNA repair mechanisms. We briefly review the impact of TLR4 and NOD1/NOD2 and their genetic variability in the process of inflammation, tumorigenesis and DNA repair, focusing in the gastrointestinal tract. We also review the available data on new therapeutic strategies utilizing TLR /NLR agonists and antagonists for cancer, allergic diseases, viral infections and vaccine development against both infectious diseases and cancer.

Naturally occurring cinnamic acid derivatives are ubiquitously distributed in the plant kingdom, and it has been proposed that their consumption contributes to the maintenance of human health. However, the molecular mechanisms underlying their health keeping effects remain unknown. In the present investigation, we evaluated the capacity of several cinnamic acid derivatives (trans-cinnamic, p-coumaric, caffeic and ferulic acids, as well as caffeic acid-methyl and -propyl esters) to protect cells from oxidative stress-induced DNA damage. It was observed that effective protection was based on the ability of each compound to (i) reach the intracellular space and (ii) chelate intracellular "labile" iron. These results support the notion that numerous lipophilic iron chelating compounds, present abundantly in plant-derived diet components, may protect cells in conditions of oxidative stress and in this way be important contributors toward maintenance of human health.

The effect of the lunate type on carpal collapse in cases of scaphoid nonunion has not been thoroughly investigated. The purpose of the present study was to determine whether any association exists or not between the capitate-triquetrum distance and occurrence of carpal collapse in cases of scaphoid nonunion. In a retrospective study, 76 patients with scaphoid nonunion formed two groups based on the capitate-triquetrum distance: forty-three patients with distance of less than 5 mm and 33 patients with distance of 5 mm or more. The two groups were comparable with respect to sex distribution, age, dominant hand involvement, manual labor, nonunion location and time from injury to final x-rays. Six patients (13.9%) in the capitate-triquetrum5 mm group and 13 patients (39.4%) in the capitate-triquetrum≥5 mm group had no signs of collapse, with significant difference (p0.05). Capitate-triquetrum distance could contribute in the decision making process for cases of scaphoid nonunion without straightforward indication for surgical intervention.

Objective: The aim of this study was to evaluate the correlation between the Manchester scale and foot pressure distribution in patients with hallux valgus deformity.Methods: The study included 152 feet of 87 patients with hallux valgus and a control group of 391 feet of 241 individuals without hallux valgus deformity. The severity of hallux valgus was determined using the Manchester scale grading system. Plantar loading patterns in 10 foot areas were determined for all participants.Results: According to the Manchester scale, 72% of the participants had no, 12.9% mild, 10.7% moderate and 4.4% severe deformity. The Manchester scale grade was highly correlated with both hallux valgus angle and first intermetatarsal angle (p=0.00). Significant differences between the four grades were present for mean pressure under the hallux and the first and second metatarsal heads only (p=0.00). The load distribution under these areas was higher as the hallux valgus progressed from mild to more severe. In all groups, the highest pressure was observed under the second metatarsal head.Conclusion: The Manchester scale was strongly associated with both the hallux valgus angle and the first intermetatarsal angle. The progression from mild to moderate and severe deformation is associated with peak pressure raise at the hallux, first and second metatarsal heads. The Manchester scale appears to be a useful tool to provide information for the degree of deformity and the pressure under painful foot areas.

The Jun family and the signal transducers and activators of transcription (STAT) are involved in proliferation and apoptosis. Moreover, c-Jun and STAT3 cooperate to regulate apoptosis. Therefore, we used double immunostaining to investigate the immunotopographical distribution of phospho-c-Jun (p-c-Jun), JunB, JunD, p-STAT3, p-STAT5, and p-STAT6 in human thymus. JunD was frequently expressed by thymocytes with higher expression in medullary compared to cortical thymocytes. p-c-Jun was frequently expressed by cortical and medullary thymic epithelial cells (TEC) and Hassall bodies (HB). p-STAT3 was frequently expressed by TEC with higher expression in cortical compared to medullary TEC and HB. p-c-Jun, JunB, p-STAT3, p-STAT5, and p-STAT6 were rarely expressed by thymocytes. JunB and JunD were expressed by rare cortical TEC with higher expression in medullary TEC. p-STAT5 and p-STAT6 were expressed by rare cortical and medullary TEC. Double immunostaining revealed p-c-Jun and JunD expression in rare CD11c positive dendritic cells. Our findings suggest a notable implication of JunD in the physiology of thymocytes and p-c-Jun and p-STAT3 in the physiology of TEC. The diversity of the immunotopographical distribution and the expression levels of p-c-Jun, JunB, JunD, p-STAT3, p-STAT5, and p-STAT6 indicates that they are differentially involved in the differentiation of TEC, thymocytes, and dendritic cells.

Jurkat cells in culture were exposed to oxidative stress in the form of continuously generated hydrogen peroxide, obtained by the addition of glucose oxidase to the medium. This treatment induced a rapid, dose-dependent increase in the ICIP (intracellular calcein-chelatable iron pool). Early destabilization of lysosomal membranes and subsequent nuclear DNA strand breaks were also observed, as evaluated by the Acridine Orange relocation test and the comet assay respectively. Somewhat later, these effects were followed by a lowered mitochondrial membrane potential, with release of cytochrome c and apoptosis-inducing factor. These events were all prevented if cells were pretreated with the potent iron chelator DFO (desferrioxamine) for a period of time (2–3 h) long enough to allow the drug to reach the lysosomal compartment following fluid-phase endocytosis. The hydrophilic calcein, a cleavage product of calcein acetoxymethyl ester following the action of cytosolic esterases, obviously does not penetrate intact lysosomal membranes, thus explaining why ICIP increased dramatically following lysosomal rupture. The rapid decrease in ICIP after addition of DFO to the medium suggests draining of cytosolic iron to the medium, rather than penetration of DFO through the plasma membrane. Most importantly, these observations directly connect oxidative stress and resultant DNA damage with lysosomal rupture and the release of redox-active iron into the cytosol and, apparently, the nucleus.

Aspects of the molecular mechanism(s) of hydrogen peroxide-induced DNA damage and cell death were studied in the present investigation. Jurkat T-cells in culture were exposed either to low rates of continuously generated H(2)O(2) by the action of glucose oxidase or to a bolus addition of the same agent. In the first case, steady state conditions were prevailing, while in the latter, H(2)O(2) was removed by the cellular defense systems following first order kinetics. By using single-cell gel electrophoresis (also called comet assay), an initial increase in the formation of DNA single-strand breaks was observed in cells exposed to a bolus of 150 microM H(2)O(2). As the H(2)O(2) was exhausted, a gradual decrease in DNA damage was apparent, indicating the existence of an effective repair of single-strand breaks. Addition of 10 ng glucose oxidase in 100 microl growth medium (containing 1.5 x 10(5) cells) generated 2.0 +/- 0.2 microM H(2)O(2) per min. This treatment induced an increase in the level of single-strand breaks reaching the upper limit of detection by the methodology used and continued to be high for the following 6 h. However, when a variety of markers for apoptotic cell death (DNA cell content, DNA laddering, activation of caspases, PARP cleavage) were examined, only bolus additions of H(2)O(2) were able to induce apoptosis, while the continuous presence of this agent inhibited the execution of the apoptotic process no matter whether the inducer was H(2)O(2) itself or an anti-Fas antibody. These observations stress that, apart from the apparent genotoxic and proapoptotic effects of H(2)O(2), it can also exert antiapoptotic actions when present, even at low concentrations, during the execution of apoptosis. Free Radic Biol Med

Human lymphocytes were exposed to increasing concentrations of SIN-1, which generates superoxide and nitric oxide, and the formation of single-strand breaks (SSB) in individual cells was determined by the single-cell gel electrophoresis assay (comet assay). A dose- and time-dependent increase in SSB formation was observed rapidly after the addition of SIN-1 (0.1-15 mM). Exposure of the cells to SIN-1 (5 mM) in the presence of excess of superoxide dismutase (0.375 mM) increased the formation of SSB significantly, whereas 1000 U/ml catalase significantly decreased the quantity of SSB. The simultaneous presence of both superoxide dismutase and catalase before the addition of SIN-1 brought the level of SSB to that of the untreated cells. Moreover, pretreatment of the cells with the intracellular Ca(2+)-chelator BAPTA/AM inhibited SIN-1-induced DNA damage, indicating the involvement of intracellular Ca(2+) changes in this process. On the other hand, pretreatment of the same cells with ascorbate or dehydroascorbate did not offer any significant protection in this system. The data suggest that H2O2-induced changes in Ca(2+) homeostasis are the predominant pathway for the induction of SSB in human lymphocytes exposed to oxidants. Free Radic Biol Med

Dendritic cells play key roles in thymic histophysiology and histopathology. Therefore, we analyzed the immunotopographical distribution of cells expressing markers of dendritic cells and macrophages in postnatal human thymus.The streptavidin-biotin peroxidise-labeled (LSAB) and the double-LSAB/alkaline phosphatase/anti-alkaline phosphatase (APAAP) immunohistochemical procedures were used.S100 protein-, Cluster of designation 1a (CD1a)-, CD207-, CD11c- and CD123-positive cells, many of them exhibiting the morphology of dendritic cells, were detected in the cortex but mainly in the medulla. These markers, except CD123, were also detected in cells of juvenile and immature Hassall bodies. CD68- and CD163-positive cells were detected in the cortex and the medulla but not in Hassall bodies.The immunohistological detection of S100-, CD1a-, CD207- and CD11c-positive dendritic cells in juvenile and immature Hassall bodies may reflect an important role of these structures in the cooperation of epithelial and dendritic cells in the process of T-cell differentiation.

Summary The combined expression patterns of cell cycle and apoptosis regulators have not been analyzed in details in human thymus to the best of our knowledge. Our objective was to provide multiparametric and combined immunohistological information regarding the expression levels and the topographical distribution of major cell cycle and apoptosis regulators in postnatal human thymus. Ki67 and cyclins A, B1, D3 and E were frequently expressed by thymocytes with higher expression in cortical than medullary thymocytes. The expression of cyclin D2 was low in thymocytes. Thymic epithelial cells (TEC) exhibited low expression of Ki67 and cyclins. Bid was frequently expressed by thymocytes, Bcl-xL by cortical thymocytes and Bcl-2 by medullary thymocytes. The expression levels of Bim and survivin in thymocytes were low. The expression levels of Bax and Mcl-1 were higher in medullary than cortical thymocytes and TEC. Bak and Bad were mainly expressed in medullary TEC and Hassall Bodies (HB). c-FLIP and Fas were frequently expressed in TEC and FasL was mainly expressed by medullary TEC and HB. Cleaved caspase-3 was expressed by scattered thymocytes at the cortex and the corticomedullary junction and very rarely at the medulla. The different expression profiles and immunotopographical distribution of cell cycle and apoptosis regulators in thymocytes and TEC indicate that their expression is tightly regulated during thymic cell differentiation and that they are differentially involved in the cell survival/death regulation of thymocytes and TEC. Furthermore, this study indicates decrease of the proliferation and caspase-dependent apoptosis of thymocytes from the cortex to the medulla.

Data regarding sources of oxidative stress in the failing myocardium are sparse. Leukocytes actively participate in the oxidative damage observed in human heart failure (HF). The intracellular labile iron pool (LIP) represents a source of toxic reactive oxygen species.We studied patients with chronic systolic HF who had a left ventricular ejection fraction (LVEF) 45%. We examined the LIP status in different populations of leukocytes in HF patients and we investigated its association with clinical and laboratory parameters, including conventional inflammatory markers.Sixty patients were finally included in the analysis (mean age: 67 ± 11 years, 54 men, 42 with ischemic cardiomyopathy). The multivariate logistic regression analysis showed that only LIP in granulocytes (OR: 0.73; 95% CI: 0.55-0.98; p=0.039) and right ventricular systolic pressure (RVSP) (OR: 0.95; 95% CI: 0.92-0.99; p=0.027) were independently associated with severe LV systolic dysfunction (LVEF30%). The correlation analysis revealed that LVEF was inversely associated with LIP in granulocytes (Spearman's rho: -0.39, p=0.002), LIP in monocytes (Spearman's rho: -0.35, p=0.007), and RVSP (Spearman's rho: -0.43, p=0.003). No significant correlation between LVEF and inflammatory indexes was noted.LIP in granulocytes is independently associated with the severity of LV dysfunction in patients with systolic HF. Intracellular redox active iron may represent a source of leukocyte reactive oxygen species in this setting.

Strong experimental data support the notion that excessive generation of reactive free radicals takes place in the ischemic area following a stroke episode and that these radicals are responsible for a substantial part of brain injury. However, the exact biochemical mechanisms underlying brain cell damage remain elusive, thus hindering the attempts for development of effective therapeutic strategies. In this presentation, the molecular mechanisms of tissue damage in relation to temporal and spatial generation of “reactive oxygen species” are considered. The mechanisms of oxidant-mediated brain damage are discussed and the role of “labile iron” in this process is evaluated. Although oxygen based oxidants are generated during all phases, their sources and intensities as well as their temporal and spatial generation differ considerably among different phases in stroke. Based on the above ­considerations, it is proposed that administration of appropriate “labile iron” chelating agents, preferentially prior to reperfusion, might improve the efficacy of any therapeutic strategy.

Publisher Summary Reactive oxygen species (ROS) are continuously generated and removed in all kinds of aerobic cells, thus creating an intracellular dynamic equilibrium that varies among different types of cells or even in the same cell type under different conditions. Increased rates of generation or decreased capacity to remove ROS leads to elevated steady-state levels of these species, a phenomenon usually assigned as “oxidative stress.” Depending on the intensity of the oxidative stress applied, basic cellular functions such as cell proliferation and differentiation are modulated, while higher levels of oxidative stress can induce transient or permanent cell arrest or even cell death either by apoptosis or necrosis. Consequently, oxidative stress has been implicated in pathogenic molecular mechanisms of a variety of diseases, including cardiovascular disease, ischemia–reperfusion syndrome, neurodegenerative diseases, cancer, and the physiological process of normal aging. Intensive research work has been performed in order to elucidate these mechanisms and to discover agents able to prevent or modulate the deleterious effects of oxidative stress that contribute to the development of these pathological complications. An unlimited number of potentially protective agents are believed to be present in diet and especially in the so-called “Mediterranean diet,” which has been shown to be able to prevent or impede the development of diseases associated with oxidative stress. This chapter provides evidence that natural phenolic compounds present ubiquitously in the Mediterranean diet and in olive oil exert their cytoprotective action mainly through chelation of intracellular redox-active iron, thus preventing the catalysis of the formation of deleterious free radicals near basic cell components, like DNA, proteins, and lipids.

Background/Aims: Oxidative damage has been reported to be involved in the pathophysiology of chronic kidney disease (CKD) as well as in the pathogenesis of cardiovascular complications of CKD patients. The aim of the present investigation was to evaluate the levels of plasma carbonyl formation, a sensitive marker of enhanced oxidative stress in predialysis, hemodialysis (HD) and peritoneal dialysis (PD) patients. Methods: Plasma samples from 20 apparently healthy control individuals and 127 CKD (stages 2, 3, 4, HD and PD) patients were evaluated by Western blot analysis for the estimation of the levels of protein carbonyl formation. Results: Albumin represented the main plasma carbonylated protein. Increasing carbonylation of albumin was detected along with the severity of CKD, reaching significance at stages 3 and 4 (p < 0.01, compared to healthy controls). The carbonylation of albumin was even higher in the plasma of HD patients (p < 0.001), while in PD patients it was not statistically significant compared to controls (p = 0.224). Conclusions: The data presented in this work indicate that oxidative stress in CKD patients gradually increased during the development of the disease. This stress is probably intensified during HD, but not in PD subjects.

Reactive oxygen species (ROS) were viewed for a long time as unavoidable by-products of normal cell catabolism. This view has recently changed and it is now apparent that ROS generation is a tightly regulated process that plays a central role in cell signaling. Thus, it is known that regulated changes in intracellular ROS levels can induce biochemical signaling processes that control basic cellular functions, such as proliferation and apoptosis which are prevalent in the development of cancer. In this short review, we will try to provide a background to this emerging field by summarizing the biochemistry of ROS-mediated cell signaling and its relation to carcinogenesis. Special emphasis will be focused on the emerging role of the so called "labile" iron (the redox-active form of iron) in ROS-mediated signaling in relation to cancer development. It is tempting to speculate that elucidation of the exact molecular mechanisms that govern ROS-mediated regulation of cell signaling will provide the basis for development of new therapeutic strategies for cancer prevention and treatment. Cancer Lett