Serum metallothionein - a potential oncomarker?

INTRODUCTION: Metallothionein's (MT) overexpression has been demonstrated immunohistochemically in neoplastic cells of many tumour types. Its elevation above the physiological level has been confirmed in circulation of their hosts. The results of studies dealing with the topic have been summarized to verify if this marker can be applied in the current oncologic practise. METHODS: The Pubmed and Google Scholar medical databases were reviewed for full-text articles focused on MT blood (plasma / serum) levels in patients with malignant tumours. RESULTS: In our review, after a precise selection, we included 8 prospective randomized trials encompassing 561 blood samples taken from patients with a large histopathological spectrum of malignancies. In general, significant differences in blood MT levels between oncological patients and healthy subjects were confirmed. No particular value of the MT level has been demonstrated to be unequivocally predictive of oncologic diseaseCONCLUSION: The results of our review suggest that although the elevation of MT in blood serum in patients with solid malignancy can be regarded as a promising tumour marker, the recommendations of its applicability in clinical practice require to be derived from further research on extended cohorts of patients (Tab. 1, Fig. 1, Ref. 49).

Prognosis of HPV-Positive and -Negative Oropharyngeal Cancers Depends on the Treatment Modality.

BACKGROUND: The association between human papilloma virus (HPV) and oropharyngeal carcinoma is a topical issue due mainly to the rapid increase in incidence over recent years. These tumors are etiopathogenetically, epidemiologically, and clinically different from other carcinomas at this location. They have a better prognosis in that they are more chemo-and radiosensitive. Indeed, this has been shown by many extensive retrospective and prospective studies. HPV status is considered an integral part of a standard histopathological examination and is included as a new biological parameter in TNM classification. MATERIALS AND METHODS: The results of 77 patients who were treated non-surgically for locally advanced oropharyngeal carcinoma at a single university ear, nose, and throat clinic were analyzed retrospectively. RESULTS: Overall and specific survival of those with HPV-positive (HPV+) tumors was better that those for HPV negative (HPV-) tumors. With the exception of TNM classification, HPV positivity appeared to be the strongest predictor of local control, and of overall and specific survival, regardless of the type of treatment. However, smoking and p53 positivity were significant negative predictors of overall survival. Patients with HPV-associated tumors had a significantly better prognosis, regardless of treatment type. The difference between treatment modalities was confirmed for the whole group of patients, but not for the HPV+ and HPV-patients specifically, most probably due to the small number of patients enrolled. CONCLUSION: The results obtained herein may constitute the first step toward the concept of treatment de-escalation in those with HPV-associated oropharyngeal carcinoma; however, this decision can be based only on the results of current extensive randomized trials. Specification of the optimal de-escalation scheme, or the choice of treatment modality, for which the difference in treatment results is most pronounced, has yet to be identified. This work was supported by grants of the Ministry of Health of the Czech Republic IGA NT12483-4/2011 and AZV 15-31627A. he authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 21. 9. 2018 Accepted: 14. 5. 2019.

Effect of ethanol and acetaldehyde at clinically relevant concentrations on atrial inward rectifier potassium current IK1: separate and combined effect.

Atrial fibrillation is the most common arrhythmia at alcohol consumption. Its pathogenesis is complex, at least partly related to changes of cardiac inward rectifier potassium currents including IK1. Both ethanol and acetaldehyde have been demonstrated to considerably modify IK1 in rat ventricular myocytes. However, analogical data on the atrial IK1 are lacking. The present study aimed to analyse IK1 changes induced by ethanol and acetyldehyde in atrial myocytes. The experiments were performed by the whole cell patch-clamp technique at 23 ± 1°C on enzymatically isolated rat and guinea-pig atrial myocytes as well as on expressed human Kir2.3 channels. Ethanol (8 - 80 mM) caused a dual effect on the atrial IK1 showing the steady-state activation in some cells but inhibition in others in agreement with the ventricular data; on average, the activation was observed (at 20 mM by 4.3 and 4.5% in rat and guinea-pig atrial myocytes, respectively). The effect slightly increased with depolarization above -60 mV. In contrast, the current through human Kir2.3 channels (prevailing atrial IK1 subunit) was inhibited in all measured cells. Unlike ethanol, acetaldehyde (3 µM) markedly inhibited the rat atrial IK1 (by 15.1%) in a voltage-independent manner, comparably to the rat ventricular IK1. The concurrent application of ethanol (20 mM) and acetaldehyde (3 µM) resulted in the steady-state IK1 activation by 2.1% on average. We conclude that ethanol and even more acetaldehyde affected IK1 at clinically relevant concentrations if applied separately. Their combined effect did not significantly differ from the effect of ethanol alone.

Increased activity of superoxide dismutase in advanced stages of head and neck squamous cell carcinoma with locoregional metastases.

The aim of the study was to investigate relationship between activity of superoxide dismutase (SOD), malondialdehyde (MDA) and tumor necrosis factor alpha (TNFalpha) and between Ala-9Val polymorphism in the gene encoding MnSOD (SOD2) and the initial stage and prognosis of the head and neck squamous cell carcinoma (HNSCC). Prospective study cohort comprised 88 patients who underwent surgical treatment for the diagnosis of HNSCC (53 patients were diagnosed with locoregional metastatic spread (N+) at the time of diagnosis). After the initial surgery subjects were followed for the subsequent period of 26 months during which 14 manifested relapse. Genotypes were detected by the PCR-based methodology. Activity of p-SOD, ery-SOD and TNFalpha were determined by ELISA, and the concentration of MDA by high performance liquid chromatography. Genotype and allele frequencies of the Ala-9Val differed neither between groups defined according to the stage of primary disease (TNM), nor between relapse vs. remission groups after the follow-up (p>0.05). Activity of p-SOD was significantly higher in T3/4 stage compared to T1/2 (p=0.01) and was also higher in N+ compared to N0 patients (p=0.002). Carriers of the Ala/Ala genotype had higher p-SOD activity (p=0.04). There was no significant difference in DFI between SOD2 genotype groups (p>0.05), however, the Ala/Ala group exhibited the shortest median DFI. In conclusion, our results suggest that increased p-SOD at the time of the initial treatment for HNSCC is connected with greater extent and nodal metastatic spread of the initial disease and with an earlier relapse of the disease. Progression of the disease might be further modified by the presence of Ala/Ala genotype of the SOD2. Activity of p-SOD could thus offer diagnostic as well as prognostic value.

Influence of indomethacin and other anti-inflammatory drugs on mobilization and production of neutrophils: studies with carrageenan-induced inflammation in rats.

The intrapleural injection of carrageenan in the rat, after a delay of 90 min, resulted in the accumulation of 100 to 300 million neutrophils in the pleural cavity, a 6-fold increase in the numbers of circulating neutrophils and the mobilization of large numbers of neutrophils from bone marrow. Thymidine labeling of bone marrow cells showed that neutrophils underwent a period of maturation (2--3 days) before release into the circulation and accumulation in the pleural cavity. Nonsteroidal anti-inflammatory drugs reduced, in a dose-dependent manner, the accumulation of white cells and fluid. Indomethacin, the most potent of the drugs tested, was equally effective whether given 30 min before or 90 min after carrageenan. This drug had no significant effect on the production or number of white cells in bone marrow, spleen or thymus. In contrast, methotrexate suppressed the inflammatory reaction only after prolonged treatment had produced almost complete depletion of cells in bone marrow and dexamethasone could be given in doses that largely depleted thymus and spleen of their white cells but which had little effect on neutrophil accumulation. It is concluded that neutrophils were mobilized from stores of mature cells in bone marrow and that indomethacin suppressed mobilization of these cells without impairing normal white cell homeostasis.

The role of cyproheptadine in the treatment of cold urticaria.

Histamine release in cold urticaria was studied before and after therapy with cyproheptadine to determine whether any effect upon histamine release could be distinguished from end-organ receptor-site blockade. Patients were asymptomatic while taking cyproheptadine, their ice cube test reverted to normal, and only one of six patients had any swelling upon ice-water submersion of one hand for 5 min. Histamine-release curves were obtained following ice-water submersion before and after cyproheptadine therapy. All patients had significant histamine release and in five of six patients there was no evident difference in the magnitude of histamine release before or after therapy. A single patient did have diminished histamine release after therapy, but could not be restudied to be sure this was not a spurious result. Our data demonstrated that cyproheptadine is extremely effective in ameliorating the symptoms and signs of cold urticaria and that its principal effect is that of an H1 receptor antagonist, thereby blocking the effects of histamine. These data further suggest that a sufficient dose of any standard antihistamine should be similarly effective and that patients who do not tolerate cyproheptadine or do not appear to respond to it should be tried on other antihistamines of the H1 type.

Evidence that histamine does not participate in carrageenan-induced pleurisy in rat.

The injection of carrageenan into the rat pleural cavity provoked an intense inflammatory reaction with the formation of an exudate which contained mainly neutrophils but which was also rich in mast cells and histamine. There was, however, no evidence that histamine participated in the reaction. The mast cells remained intact, and no increase in extracellular histamine levels was observed. Prior treatment with bot H1 and H2 histimine receptor antagonists or depletion of the histamine stores by pretreatment with compound 48/80 did not alter the reaction. In contrast, the exudate formed in response to the intrapleural injection of small doses (0.05 mg/kg) of compound 48/80 was reduced by pretreatment with the antihistamine compounds and, unlike the exudate formed after carrageenan injection, was devoid of neutrophils. Since saline washes of the pleural cavity of untreated rats had histamine and mast cell contents similar to those of the exudates of the carrageenan-treated rats, the source of histamine appeared to be mast cells from the pleural cavity.

Evaluation of a patient with cold and cholinergic urticaria.

A-20-year-old male Army paratrooper presented with a history of inducible urticaria associated with exercise as well as cold exposure. Upon evaluation, he not only had a positive ice cube test, but also had a positive mecholyl skin test with numberous satellite lesions and generalized punctate urticaria following exercise challenge. Thus, he appeared to have combined cold and cholinergic urticaria. When mediator release was examined during cold and exercise challenge, histamine release was observed in each instance; a rapid rise and fall of plasma histamine was seen after cold challenge, while a lag phase followed by sustained elevation of plasma histamine was associated with exercise challenge. This represents the fourth reported case of combined cold and cholinergic urticaria and is the first in whom mediator release was assessed. The time-course of histamine release was characteristic of each disorder.

Blood and urine histamine levels in normal and pathological states as measured by a radiochemical assay.

The utility of the enzymatic radiochemical assay of histamine in diagnosing diseases with known abnormalities in histamine production was investigated. Whole blood histamine levels were abnormal only in patients with basophilia, i.e. chronic myelocytic leukemia or polycythemia vera. Histamine was not detectable (less than 1 ng/ml) in normal plasma but was detected in plasma of some patients witheither mastocytosis or chronic myelocytic leukemia. These patients also had symptoms which could be attributed to histamine release as, for example, hyperchlorhydria and hypotension. Urinary histamine excretion was also abnormally high in these diseases compared to normal subjects (range less than 5-42 microgram/24 h, n = 31). Patients with systemic mastocytosis had higher urine values (greater than 150 microgram/24 h) than those with cutaneous mastocytosis (39-88 microgram/24 h), and the urinary histamine excretion appeared to be an index of the severity of the diseases. Studies with L-histidine loading suggest that the kidney is one possible source of urinary histamine.

Antiproliferative activity of anti-inflammatory drugs in two mammalian cell culture lines.

The nonsteroid anti-inflammatory drugs inhibited cell proliferation when added to rat hepatoma and human fibroblast cultures. The inhibition was reversible; normal growth resumed when the cultures were washed free of drug. Protein and nucleic acid synthesis, as measured by isotope incorporation was also reduced, although this reduction was probably a reflection of the decrease in cell numbers. An exception was that, in low concentration, the salicylate drugs, salicylamide, salicylic acid and aspirin, stimulated protein and nucleic acid synthesis, but in high concentrations (greater than 1 mM) they inhibited culture growth as well as protein and nucleic acid synthesis. Pharmacologically inactive derivatives, such as m-hydroxybenzoic acid and gentisic acid, were not inhibitory in concentrations up to 5 mM. The order of potency in inhibiting culture growth, meclofenamate greater than indomethacin greater than salicylamide greater than phenylbutazone greater than phenacetin greater than aspirin = salicylic acid, was similar to that reported for their anti-inflammator activity and their ability to inhibit prostaglandin synthesis. The antiproliferative activity of these drugs may, in part, account for their anti-inflammatory and toxic actions in vivo.

Interference with histamine and imidazole acetic acid metabolism by salicylates: a possible contribution to salicylate analgesic activity?

In man, rats and mice, the urinary excretion of the histamine and L-histidine metabolite, imidazole acetic acid, is increased and that of the conjugated metabolite, ribosylimidazole acetic acid, decreased by small doses of salicylates. In contrast to salicylates, other non-salicylate anti-inflammatory drugs, indomethacin, phenylbutazone, phenacetin and acetaminophen do not influence the excretion of the urinary metabolites of histamine and L-histidine. Since imidazole acetic acid is reported to have analgesic and narcotic activity, there is the inference that the analgesic properties of salicylate might be due in part to interference in imidazole acetic acid metabolism.

Alteration of tumor growth by aspirin and indomethacin: studies with two transplantable tumors in mouse.

Oral daily administration of aspirin or indomethacin retarded growth of experimental tumors in mouse. Aspirin treatment, 150 mg/kg twice daily, inhibited growth of a transplantable mast-cell ascites tumor (P815) by 39-43% (p less than 0.001) and of a s.c. transplanted Lewis lung carcinoma by 52% (p less than 0.025) without adversely affecting body growth. The total serotonin, histamine and histidine decarboxylase content of the ascites tumor was also reduced as was the urinary excretion of the amines. Treatment with 3 and 5 mg/kg indomethacin resulted in 40% (p less than 0.01) and 80% (p less than 0.001) reduction, respectively, in ascites tumor growth. With the higher dose of indomethacin, no tumor was observed in half of the animals inoculated with tumor, although signs of indomethacin toxicity (reduced body growth, gastric lesion) was evident in the animals.

The role of histamine in burn, tourniquet and endotoxin shock in mice.

The release of histamine and mortality was studied in mice after various types of experimental shock. In burn shock, serum histamine rose significantly after injury, but there was no correlation between increased serum histamine and high mortality as a consequence of several therapy regimens. For example, after treatment with histamine or Compound 48/80 before burning, there was a rise of serum histamine, yet shock mortality fell significantly. Although separate administration of antagonists of H1 - or H2 - histamine receptors had no effect on mortality, pretreatment with both diphenhydramine and burimamide significantly increased shock mortality. In tourniquet shock, serum histamine rose significantly, and treatment with both antagonists before trauma produced a significant elevation of shock mortality. In endotoxin shock, prior treatment with one or both drugs did not change mortality. These results suggest that endogenous histamine is not a lethal factor in burn and tourniquet trauma, but rather it appears to have a compensatory, beneficial effect.

In vivo studies of mediator release in cold urticaria and cholinergic urticaria.

Six patients with cold urticaria were found to possess elevated plasma histamine levels after cold challenge by placing one hand in ice water for 4 minutes. A single patient became hypotensive during the procedure and had a level of 260 ng/ml. histamine in the venous effluent from his hand. No elevation of plasma serotonin or bradykinin was observed. Two patients with cholinergic urticaria possessed elevated plasma histamine levels during and after vigorous exercise for 10 minutes; these patients also gave a positive test for vibration-induced angioedema. A single patient with cholinergic urticaria possessed elevated baseline serotonin levels and elevated levels during and after exercise but no elevation of plasma histamine or bradykinin. The results suggest that histamine is the major mediator of urticaria and hypotension in cold urticaria. Histamine also appears to be released coincident with the development of urticaria in some patients with cholinergic urticaria, while elevated serotonin levels in a single atypical patient suggest that a subpopulation of patients with cholinergic urticaria possess a different pathogenesis.