Somatic genomic imbalances in 'tumour-free' surgical margins of oral cancer.

Up to 30% of oral squamous cell carcinoma (OSCC) patients develop local recurrence and distant metastasis. The molecular status of histologically cancer-free tumour margins could be a critical factor in predicting tumour behaviour. The aim of this study was to detect somatic genomic imbalances in OSCC with emphasis on the surgical margins. DNA was isolated from tumour tissues, margin tissues, and blood samples (used as control) obtained from 11 OSCC patients, and genome-wide array comparative genomic hybridization was performed. Imbalances were present in both tumours and margins, although, as expected, they were more prevalent in tumours (duplications, P = 0.0002; deletions, P = 0.0001). Duplications were more frequent than deletions in both tumours and margins, but without statistical significance. Fifteen imbalances in tumour tissues were recurrent and all of them were duplications. Four of these were found both in tumours and margins and involved chromosomes 1q, 8p, Xp, Yp, and Yq. Four imbalances were recurrent in margin tissue and all of them were duplications (autosomes 8 and 17 and both sex chromosomes). Histologically 'cancer-free' margins hide genomic alterations consistent with unexplained OSCC recurrences. Establishing the molecular status of the margins could improve outcome prediction.

Reproductive immunology in viviparous mammals: evolutionary paradox of interactions among immune mechanisms and autologous or allogeneic gametes and semiallogeneic foetuses.

Literally, reproductive immunology was born in bovine on-farm reproduction where seminal experiments intended for developing methods for embryo transfer in cattle were performed. Actually, these experiments led to two of major concepts and fundamental principles of reproductive immunology using the bovine species as a model for biomedical research, namely the concept of acquired immunological tolerance and the paradox of the semiallogeneic bovine foetus whereby such organism can develop within an immunologically competent host. Peter Medawar, a scientist who together with Frank Macfarlande Burnet shared the 1960 Nobel Prize in physiology or medicine for discovery of acquired immunological tolerance, while studying dizygotic cattle twins, thereby giving birth to reproductive immunology. Also, these findings significantly influenced development of organ transplants and showed that using farm animals as models for studying transplantation immunology had general relevance for mammalian biology and health including those of humans. However, the interest for further research of the fascinating maternal immune influences on pregnancy and perinatal outcomes and of the prevention and treatment of immunologically mediated reproductive disorders in viviparous mammals of veterinary relevance by veterinary immunologists and reproductive clinicians have been very scarce regarding the application of nonspecific immunomodulatory agents for prevention and treatment of subfertility and infertility in pigs and cattle, but still broadening knowledge in this area and hold great potential for improving such therapy in the future. The aim of the current overview is to provide up-to-date information and explaining/translating relevant immunology phenomena into veterinary practice for specialists and scientists/clinicians in reproduction of animals.

Likely Transiting Exocomets Detected By Kepler.

We present the first good evidence for exocomet transits of a host star in continuum light in data from the Kepler mission. The Kepler star in question, KIC 3542116, is of spectral type F2V and is quite bright at Kp = 10. The transits have a distinct asymmetric shape with a steeper ingress and slower egress that can be ascribed to objects with a trailing dust tail passing over the stellar disk. There are three deeper transits with depths of ≃ 0.1% that last for about a day, and three that are several times more shallow and of shorter duration. The transits were found via an exhaustive visual search of the entire Kepler photometric data set, which we describe in some detail. We review the methods we use to validate the Kepler data showing the comet transits, and rule out instrumental artefacts as sources of the signals. We fit the transits with a simple dust-tail model, and find that a transverse comet speed of ∼35-50 km s-1 and a minimum amount of dust present in the tail of ∼ 1016 g are required to explain the larger transits. For a dust replenishment time of ∼10 days, and a comet lifetime of only ∼300 days, this implies a total cometary mass of ≳ 3 × 1017 g, or about the mass of Halley's comet. We also discuss the number of comets and orbital geometry that would be necessary to explain the six transits detected over the four years of Kepler prime-field observations. Finally, we also report the discovery of a single comet-shaped transit in KIC 11084727 with very similar transit and host-star properties.

Genetic and epigenetic alterations in the tumour, tumour margins, and normal buccal mucosa of patients with oral cancer.

Despite adequate surgical resection, oral squamous cell carcinoma (OSCC) shows a high rate of recurrence and metastasis, which could be explained by the presence of molecular alterations in seemingly normal tumour margins and the entire oral mucosa. The aims of this study were (1) to assess the presence of gene amplification (c-Myc and HER2) and promoter methylation (p14 and p16) in the tumours, tumour margins, and unaffected oral mucosa of 40 OSCC patients, and (2) to evaluate the possibility of using these alterations as prognostic markers. c-Myc and HER2 genes were quantified by means of real-time PCR (qPCR), and p14 and p16 methylation status was determined by methylation-specific PCR (MSP PCR). All tissues examined exhibited molecular alterations in various proportions. Tumour tissues, as expected, showed the highest prevalence of alterations, while oral mucosa showed the lowest. Multiple alterations (co-alterations) in tumours and tumour margins were significantly more frequent than in unaffected oral mucosa (P<0.001 and P=0.027, respectively). HER2 amplification in margin tissue (P<0.001) and swabs (P=0.013), as well as the existence of three co-alterations in margins (P=0.001) and macroscopically unaffected oral mucosa (P<0.001) were correlated with shorter disease-specific survival.

Q fever outbreak in the village of Nocaj, Srem County, Vojvodina Province, Serbia, January to February 2012.

From 27 January to 10 February 2012, a total of 43 cases of Q fever were notified in the village of Nocaj, Srem county, Autonomous Province of Vojvodina, Republic of Serbia. Q fever was laboratory confirmed in 37 notified cases. Alhough, the outbreak is considered over, the outbreak investigation is still ongoing in order to identify aetiologic factors relevant for this outbreak.

Effect of gut active carbohydrates on plasma IgG concentrations in piglets and calves.

Improving immune status in neonates is crucial to health and production. Gut active carbohydrates (GAC) have been associated with increasing immunoglobin levels and immonucompetence development in mammals. The objective of the following studies was to evaluate whether GAC (mannan-oligosaccharides) applied orally to progeny immediately following parturition, improved blood plasma immunoglobulin (Ig) type G concentrations in piglets and calves. Three trials were conducted comparing control groups with those receiving GAC orally. The first two trials used piglets that were monitored for blood IgG at 2 days of age and for changes in body weight (BW), and the third trial monitored calf IgG from birth to 21 days of age. Piglets in the experimental group received 0.75 g GAC in 10 ml saline at birth and 24 h of age. The calf trial compared the control group against calves that received 22.5 g GAC mixed into 4.5 l of colostrum (to give 5 g/l) in the first 24 h after parturition. Blood serum samples were taken at 2 days post partum in piglets, and at several time points from 6 h to 21 days of age in calves, and were analysed for IgG levels by radial immunodiffusion. In the first piglet trial, significantly higher levels (32%) of IgG were observed for piglets fed GAC (P < 0.001), and in the second, IgG concentration was elevated by 23% (P < 0.01) and BW increased by 9% (P = 0.023) with GAC supplementation. Significant improvements for calves were recorded at all time points in those fed GAC (P < 0.05), with an increase in serum IgG observed after the first day, which was maintained throughout the sampling period, resulting in a difference of 39% at the end of the trial (21 d). These findings form a basis for further studies, which are required to investigate possible modes of action involved in enhancing blood immunoglobulin concentrations in young animals, and the longer-term effects this may have on the development of the immune response.

Effects of peroral insulin and glucose on circulating insulin-like growth factor-1, its binding proteins and thyroid hormones in neonatal calves.

There is disagreement in the literature about the ability of neonatal calves to absorb perorally administered insulin. This study evaluated the absorption of a bolus of insulin administered alone or with an energy souce and its effects on the circulating insulin-like growth factor system and thyroid hormones in newborn Holstein-Friesian calves. Within 1 h of dosing, mean serum insulin and triiodothyronine (T3) concentrations had increased considerably, whether the insulin was applied alone (n = 4) or together with glucose (n = 4), accompanied by marked hypoglycemia. No significant changes were observed in control calves (n = 4) given the vehicle solution. Increased serum glucose and T3 concentrations with no change in insulinemia occurred in a 4th group of calves given glucose alone. At 32 h of age and after 3 meals of colostrum there were no differences in glycemia, insulinemia, or proteinemia among the 4 groups of calves examined. Mean serum insulin-like growth factor-I (IGF-I) tended to decrease over this period in the control group. The decrease was more pronounced in the insulin-treated group but absent in both groups that received glucose. These differences were associated with equivalent differences in abundance of the 40-45K IGF-binding protein-3 (IGFBP-3); however, lower molecular mass IGFBPs were not affected. The results show that a pharmacological peroral dose of insulin can lead to rapid systemic alterations in the IGF/IGFBP system in neonatal calves that can be modified by simultaneous administration of a small energy supply in the form of glucose.

Redundancy problem in writing: from human to anthropomorphic robot arm.

This paper presents the analysis of motion of a redundant anthropomorphic arm during the writing. The modeling is based on the separation of the prescribed movement into two motions: smooth global, and fast local motion, called distributed positioning (DP). The distribution of these motions to arm joints is discussed. It is based on the inertial properties and actuation capabilities of joints. The approach suggested allows unique solution of the inverse kinematics of redundant mechanisms such as human arm and anthropomorphic robot arm. Distributed positioning is an inherent property of biological systems. Humans, when writing, as shown in literature and in our earlier work control their proximal joints, while the movement of distal joints follow them (synergy). To enhance capabilities of robots, new control schema are necessary. We show that robot control can be improved if it is biological analog. The major aim of this study is to promote such a hypothesis by using anthropomorphic robot arm in writing as an example.

[Disorders of phosphate, calcium and magnesium ion metabolism in chronic renal insufficiency]. / Poremacaj metabolizma jona fosfata, kalcijuma i magnezijuma u hronicnog insuficijenciji bubrega.

In hundred patients with different stages of renal functional damage, including the controls of healthy persons we examined the disturbances of phosphate, calcium and magnesium ion metabolism during chronic renal failure (CRF) progression. We also determined the serum levels of regulatory substances, parathormone and 1.25 dihydroxyvitamin D3 in 43 pts. grouped according to their group, sex, age average and season (autumn-winter) distribution. We established actual renal compensatory mechanisms for all of substances we determined, which were involved in homeostasis preservation of those substances. Some characteristic changes in plasma substance concentration and renal mechanisms of their regulation during CRF progression, suggest the following conclusions. No substances have preserved body homeostasis, although renal compensatory mechanism were expressed by different size for any of them. Those disturbances very tightly correlate with the stage of renal functional deterioration, as well as the serum parathormone rising and serum 1.25 DHHC decreasing.

[Pathogenic mechanisms of importance for the development of secondary hyperparathyroidism in chronic renal insufficiency]. / Mogucni patogenetski mehanizmi od znacaja za razvoj sekudnarnog hiperparatireoidizma u hronichnoj insuficijenciji bubrega.

Secondary hyperparathyroidism is a complex pathophysiologic event which we examined in hundred patients Sixteen of them belonged to the control with normal renal function, and the rest of them were categorised into four subgroups according to the actual level of chronic renal failure, determined by GFR a.i. by endogenous creatinine clearance. We determined the serum level of parathormone (C-PTH) and plasma concentration of phosphate and calcium in all of patients. In 43 of them we measured serum level of 1.25 dihydroxycholecalcipherol and ionised calcium. Our results suggested an important parallelism between parathormone hypersecretion (GFR +/- 58.3 ml/min), 1.25 dihydroxycholecalcipherol hypovitaminosis (GFR +/- 50.7 ml/min) and hypocalcemia (GFR +/- 47.7 ml/min) which connect them in very strong relationship. Even if secondary hyperparathyroidism in chronic renal failure is caused by numerous factors, it appears that the hypovitaminosis of 1.25 dihydroxycholecalcipherol with consequent hypocalcemia and phosphate retention acts as a main movement of this complex pathophysiology mechanism.

[Levels of dihydroxyvitamin D3 and hydroxyvitamin D in patients with chronic renal insufficiency]. / Koncentracije dihidroksivitamina "de-tri" i hidroksivitamina "de" kod bolesnika s hronicnom insuficijencijom bubrega.

According to the aim of our research to find a regulatory of changes of the serum levels of 1.25 dihydroxyvitamin D3 and 25 OH D vitamin, we examined their serum concentration in 43 among 100 pts. with normal and impaired renal function with different stages of chronic renal failure (CRF). In all of 100 pts. we measured plasma phosphate and calcium concentration and endogenous creatinine clearance. The results of our research showed homeostasis abnormalities in all of parameters we examined. We found out that the serum 1.25 (OH)2 D3 levels decreased in early course of CRF, hypovitaminosis occurred when glomerular filtration rate reaches values equal or less than 50.7 ml/min. Hypovitaminosis of 25 OH D expressed in end-stage of CRF, while hyperphosphatemia was commonly seen in moderate CRF, when GFR became less than 28.6 ml/min. Hypocalcemia obtained when GFR was equal or less than 47.7 ml/min.

Reduction of cortisol levels after single intra-articular and intramuscular steroid injection.

BACKGROUND: This study was undertaken to determine the influence of single intra-articular or intra muscular injections of methylprednisolone acetate on the hypothalamic-pituitary-adrenal axis. PATIENTS AND METHODS: Twenty-one patients with rheumatic disease who had never been treated with systemic glucocorticoids and had not received local injections of these agents for the preceding 2 months, were given 40 mg of methylprednisolone acetate. Group I (11 patients) received one intra-articular injection into the knee, and Group II (10 patients) received the same dose intramuscularly. RESULTS: In Group I, serum cortisol levels were significantly decreased 24 hours after injection (228.2 +/- 8.7 nmol/L versus 193 +/- 16.3 nmol/L; P < 0.05). Serum cortisol levels were decreased in 9 of the 11 patients, by an average of 21.5%. Two patients' levels were below 138 nmol/L, which is considered to be the lower limit of normal range. Serum cortisol levels were below normal range in 3 patients 72 hours after intra-articular steroid injection. In Group II, serum cortisol levels were significantly decreased at 72 hours after injection (239.6 +/- 10.3 nmol/L versus 175.6 +/- 21.4 nmol/L; P < 0.01). Three patients' levels were below normal. By 72 hours postinjection, serum cortisol concentrations in 9 of 10 patients were decreased by an average of 31% compared to preinjection values. CONCLUSION: The present study suggests that decreased adrenocortical secretion, as reflected in depressed cortisol levels, can result from a single, low-dose, intra-articular or intramuscular injection of depot corticosteroids.

Immunomodulatory effects of extracellular secretory vesicles isolated from bovine semen.

We demonstrate that extracellular secretory vesicles isolated from bovine seminal plasma have immunomodulatory properties. They inhibit mitogen induced proliferation of bovine and human peripheral blood lymphocytes in a dose dependent fashion. They also inhibit phagocytosis of latex particles by bovine neutrophils. Phagocytosis of opsonised Staphylococcus aureus however was not affected. Furthermore phorbol ester and chemotactic peptide induced superoxide production was decreased especially when a suboptimal dose of stimulants was used. We suggest that extracellular secretory vesicles may preserve sperm survival in the female reproductive tract.

Synthetic peptides as probes for G protein function. Carboxyl-terminal G alpha s peptides mimic Gs and evoke high affinity agonist binding to beta-adrenergic receptors.

The molecular interfaces between Gs and the beta-adrenergic receptor were investigated using synthetic peptides corresponding to various regions of its alpha subunit, alpha s. These experiments were carried out on saponin-permeable C6 glioma cells in which the beta-adrenergic receptor appears tightly coupled to Gs. Synthetic site-specific peptides from alpha s (corresponding to amino acids 15-29, 354-372, and 384-394) and alpha i (8-22, 315-324, and 345-455) were tested for their ability to interfere with coupling between the beta-adrenergic receptor and Gs. The two carboxyl-terminal peptides from alpha s blocked beta-adrenergic stimulation of adenylyl cyclase in permeable cells. However, only alpha s-354-372 had this effect in C6 membranes. It is suggested that the partial uncoupling of Gs, which occurs subsequent to cell disruption, may be related to a change in the interaction of the alpha s carboxyl terminus with the beta-adrenoreceptor. Two carboxyl-terminal peptides, 354-372 and 384-394, could also mimic the effect of Gs to increase agonist affinity for the beta-adrenergic receptor. In combination, alpha s-354-372 and alpha s-384-394 increased the ability of isoproterenol to compete with 125I-pindolol binding in a partially additive manner. Synthetic peptides from alpha i and amino-terminal peptides from alpha s had no effect on beta-agonist binding, suggesting a high specificity of peptide effects. Two findings suggest that these peptides bind directly to the beta-adrenergic receptor and stabilize its high agonist affinity conformation. First, GTP and hydrolysis-resistant GTP analogs did not alter the high affinity binding in the presence of high concentrations of the peptides. Second, in S49 lymphoma cyc- cells, which lack Gs, these peptides evoked the high affinity agonist binding state of the beta-receptor. Neither peptide had an effect on antagonist binding affinity, as measured by propranolol displacement of 125I-pindolol. These data suggest that at least two regions on the alpha subunit of Gs participate in high affinity Gs binding to the beta-adrenergic receptor. The fact that these small peptides could mimic the holo-Gs effect on the receptor is rather surprising, and the specificity of the effect suggests that the primary and secondary structure of small regions of alpha s contain much of the information for specific interaction with beta-adrenergic receptors.

Cholesterol crystals in synovial and bursal fluid.

Cholesterol crystals were found in two patients with classic rheumatoid arthritis (RA). In one patient, cholesterol crystals were found in synovial fluid from both shoulder joints, and in the second they were in an olecranon bursa. To examine the possible systemic etiology of cholesterol crystals in synovial and bursal fluid, lipid concentrations and the presence of serum antilipoprotein antibodies were measured. Antilipoprotein antibodies were not found. The concentration of lipid and lipoproteins, as well as the normal pattern of lipoprotein on agarose gel, eliminates the possibility of hyperlipoproteinemia. Results seemed to exclude a systemic etiology for the formation of cholesterol crystals in synovial and bursal fluid in the RA patients. It appears that several local factors such as defective drainage, local destruction, increased permeability of synovial membrane, and intraarticular (bursal) bleeding are possible etiologies.

Systemic lupus erythematosus and Hodgkin's lymphoma.

OBJECTIVE: Systemic lupus erythematosus (SLE) is associated with lymphoproliferative diseases such as Hodgkin's lymphoma. Since there is considerable overlap between the features of SLE and Hodgkin's lymphoma (SLE/HL) there can be a great difficulty in diagnosing Hodgkin's lymphoma in the presence of SLE. Our objective was to determine the features that can suggest coexistent SLE/HL: METHODS: We describe 3 cases of Hodgkin's lymphoma that occurred in the setting of SLE and review the features of 11 reported cases in which the diseases occurred simultaneously. We compare the features of these cases with those of Hodgkin's lymphoma and SLE occurring alone. RESULTS: Mediastinal adenopathy was seen in 58% and retroperitoneal adenopathy in 17% of patients with SLE/HL, while it was seen only rarely in these sites in cases with SLE. Severe generalized pruritus, which occurs in only 2.8% of cases of SLE, was seen in 63% of cases with SLE/HL: Features such as fever, weight loss, peripheral lymphadenopathy, splenomegaly and hepatomegaly are common to both Hodgkin's lymphoma and SLE. Renal disease is seen in half of the cases of SLE. It is rare in Hodgkin's lymphoma where, unlike SLE, it is only of minimal change or membranous type. Arthritis and positive antinuclear antibodies are rare in Hodgkin's lymphoma. CONCLUSIONS: Persistent lymphadenopathy, especially mediastinal and/or retroperitoneal lymphadenopathy, eosinophilia and generalized pruritus in a patient with SLE not responding to treatment may be indicative of coexistent Hodgkin's lymphoma.

Myasthenia gravis and scleroderma.

Myasthenia gravis is an autoimmune disease that is associated with antibodies to acetylcholine receptors. It is associated with other autoimmune diseases such as thyroiditis (10%), systemic lupus erythematosus (2-8%), rheumatoid arthritis (4-7%), Sjögren's syndrome and polymyositis. It is not commonly found with scleroderma. We describe a case of scleroderma developing in a patient 6 years after the onset of myasthenia gravis. HLA-B8/DR3 may play a role in the association of the 2 conditions.

Antilipoprotein antibodies in rheumatoid arthritis.

Antibodies against very low-density lipoproteins and low-density lipoproteins (aLA) were found in 26 of 69 (38%) patients with active rheumatoid arthritis (RA) but not in any control subjects (ie, 40 patients with psoriatic arthritis, 21 patients with osteoarthritis, and 65 healthy blood donors). In 21 RA patients (30%), lipoproteins were found in the dissociated components of circulating immune complexes. RA patients with aLA had significantly decreased cholesterol levels in all lipoprotein fractions and total serum lipids, while serum triglycerides were significantly increased compared with RA patients without aLA. Anticardiolipin antibodies as measured by the Venereal Disease Research Laboratory test were not found in any subject in this study. These findings suggest a possible autoimmune origin of dyslipoproteinemia in some patients with active RA.