Essential Oil Constituents as Anti-Inflammatory and Neuroprotective Agents: An Insight through Microglia Modulation.

Microglia are key players in the brain's innate immune response, contributing to homeostatic and reparative functions but also to inflammatory and underlying mechanisms of neurodegeneration. Targeting microglia and modulating their function may have therapeutic potential for mitigating neuroinflammation and neurodegeneration. The anti-inflammatory properties of essential oils suggest that some of their components may be useful in regulating microglial function and microglial-associated neuroinflammation. This study, starting from the ethnopharmacological premises of the therapeutic benefits of aromatic plants, assessed the evidence for the essential oil modulation of microglia, investigating their potential pharmacological mechanisms. Current knowledge of the phytoconstituents, safety of essential oil components, and anti-inflammatory and potential neuroprotective effects were reviewed. This review encompasses essential oils of Thymus spp., Artemisia spp., Ziziphora clinopodioides, Valeriana jatamansi, Acorus spp., and others as well as some of their components including 1,8-cineole, ß-caryophyllene, ß-patchoulene, carvacrol, ß-ionone, eugenol, geraniol, menthol, linalool, thymol, α-asarone, and α-thujone. Essential oils that target PPAR/PI3K-Akt/MAPK signalling pathways could supplement other approaches to modulate microglial-associated inflammation to treat neurodegenerative diseases, particularly in cases where reactive microglia play a part in the pathophysiological mechanisms underlying neurodegeneration.

Differences in Histological Subtypes of Invasive Lobular Breast Carcinoma According to Immunohistochemical Molecular Classification.

The technical complexity of gene expression profiling in routine practice has necessitated the use of surrogate molecular classification of breast cancer, based on immunohistochemical analyses. BACKGROUND AND OBJECTIVES: The aim of this study was to compare the differences between histological and molecular subtypes of invasive lobular carcinoma (ILC) of the breast, in order to be able to predict the behavior and prognosis of the disease, as well as to effectively determine therapy. MATERIAL AND METHODS: This study included 263 cases of breast ILC diagnosed over a seven-year period. The diagnosis of invasive lobular carcinoma is based on the characteristic growth pattern and phenotype of cancer cells with the respective subtypes: classic, alveolar, solid, tubulolobular, pleomorphic and mixed lobular type. The examined cases were divided into five groups according to molecular classification based on the expression of ER, PR, HER2 and Ki67 immunohistochemical markers. RESULTS: It was found that the pleomorphic subtype occurred statistically significantly less often as the luminal A subtype compared to others (p = 0.00027), and the HER2-enriched subtype occurred statistically significantly more often in the pT4 stage (p = 0.024). CONCLUSIONS: The results of this study significantly singled out the luminal A subtype, and among them classic ILC, as the subtype with the most favorable expression ratio of the investigated predictive/prognostic immunohistochemical markers.

Constituents of Bupleurum praealtum and Bupleurum veronense with Potential Immunomodulatory Activity.

In this investigation, chromatographic separations of the diethyl ether extracts of two European annual Bupleurum taxa, B. praealtum and B. veronense, yielded nine new natural products, namely, a series of esters of stereoisomeric tetradeca-5,7,9,11-tetraen-1-ols (1-4 and 8), a tetra-unsaturated γ-tetradecalactone (5), a dibenzylbutyrolactone lignan (7-oxoarcitin, 6), a falcarinol-related 17-membered macrolide (7) possessing a conjugated diyne-system, and an acylphloroglucinol derivative (9). All these new compounds were fully characterized by NMR, IR, UV, MS, and optical rotation measurement, including 1H NMR full spin spectral simulation, whereas the absolute configurations of 1, 5, and 9 were determined via chemical correlations and NMR analysis of Mosher esters. The in vitro potential immunomodulatory activities of 1, 4, 5, and (+)-arcitin were assessed by determining their effects on the functional properties of isolated rat splenocytes and peritoneal macrophages. The results obtained support the known immunomodulatory ethnomedicinal usage of Bupleurum species.

Immunomodulatory activity of marine natural products: Synthesis, spectral characterization and toxicity assessment of natural and related synthetic iodinated tyramides.

The toxicity of natural marine iodoarenes or their synthetic counterparts is widely unknown despite the fact that triiodothyronine and thyroxine are members of this class. In this work we aimed to expand such knowledge on iodinated marine natural products and tested an ascidian (Didemnum rubeum) metabolite, N-(3,5-diiodo-4-methoxyphenethyl)benzamide, together with closely related synthetic iodinated tyramides: N-(2,5-diiodo-4-methoxyphenethyl)benzamide, N-(3-iodo-4-methoxyphenethyl)benzamide, N-(4-methoxyphenethyl)benzamide, and N-(3-iodo-4-methoxyphenethyl)formamide, for their effect on the viability of rat macrophages, as well as acute toxicity on Artemia salina. The tested tyramides exerted a varying degree of toxicity towards brine shrimps, but in certain cases, the determined lethal concentrations were even lower than those of known toxicants (e.g. strychnine sulfate, SDS). The toxicity was highly dependent on the structure of these mutually related compounds, while the natural one was shown to be the most toxic. In the case of macrophage cultures, the tested tyramides exerted much less toxicity but were found to have an effect on the functioning of these normal immune cells. The samples of the tyramides were obtained by synthesis, and were fully structurally and spectrally characterized, which also provided corroboration of the proposed structure of the natural product originally isolated in minute amounts.

Toxic essential oils: anxiolytic, antinociceptive and antimicrobial properties of the yarrow Achillea umbellata Sibth. et Sm. (Asteraceae) volatiles.

Many plant species are used for medicinal purposes without the knowledge of their possible toxic effect. The ethnopharmacologically renowned genus Achillea L. (Asteraceae) is even more troublesome in this respect since different taxa are believed to have the same beneficial properties as A. millefolium. According to the median lethal i.p. dose (LD(50)=853 mg/kg, mice), the volatiles of Achillea umbellata Sibth. et Sm. are more toxic than the thujone-containing essential oils (LD(50)>960 mg/kg). A GC-MS analysis of A. umbellata oil revealed the presence of a series of fragranyl esters (six new natural products). The major constituents of this oil, the rare monoterpene alcohol fragranol and fragranyl acetate, and one more ester (benzoate), as well as the oil itself, showed antianxiety, analgesic and, in some instances, paralyzing properties at 50-150 mg/kg but these are very likely sign of intoxication and not of possible beneficial effects of the plant volatiles. Testing of antimicrobial activity demonstrated that the oil possesses moderate activity against pathogenic microorganisms, but the effect of the oil differs in pro- and eukaryotic cells. According to the results obtained, fragranol may be considered as the main active principle responsible for the observed activity/toxicity.

An approach to malignant mammary phyllodes tumors detection.

BACKGROUND/AIM: Mammary phyllodes tumors (MPT) are uncommon fibroepithelial (biphasic) neoplasms whose clinical behavior is difficult to predict on the basis of histological criteria only. They are divided into benign, borderline malignant and malignant groups. Sometimes it appears difficult to distinguish these tumors from other types of soft tissue sarcomas. Because of the relatively scant data on the role of biological markers in MPT histogenesis, we have decided to undertake the following study, trying to shed more light on the issue by investigating the following elements that make up MPT: their histological patterns, biological behavior, enzymohistochemical, histochemical and immunohistochemical characteristics (ICH) together with the mast cell analysis. METHODS: We examined the biopsy material of 35 MPT in our laboratory. Enzymohistochemistry was performed on frozen sections (method of Crowford, Nachlas and Seligman). The used methods were classical hematoxylin-eosin (H & E); histochemical Massontrichrome, Alcian-blue, Periodic acid Schiff and immunohistochemical LSAB2 method (DacoCytomation). Ki-67, c-kit, vimentin, estrogen receptor (ER), progesterone receptor (PR) and Her-2 oncoprotein immunohistochemistry was performed on all tumors. RESULTS: The patients were ranged per age from 30--62 years (mean 43.3 years, median 39 years). A total of 35 cases of MPT were included: 20 benign (57%), 6 borderline malignant (17%) and 9 malignant (26%). Twenty-two patients (62.8%) underwent segmental mastectomy, while 13 (37.2%) had total mastectomies. Twenty-eight patients had negative surgical margins at original resection. The mean size of malignant MPT (7.8 cm) was larger than that of benign MPT (4.5 cm). Significant features of the malignant MPT were: stromal cellularity, stromal cellular atypism, high mitotic activity, atypic mitoses, stromal overgrowth, infiltrative tumor contour and heterologous stromal elements. Benign MPT showed strong enzymohistochemical Leucine Amino Peptidase (LAP) activity in both epithelial and stromal components while it was weak or absent in the epithelial parts of the malignant tumors. Acid mucopolysacharides were present in the stromal component of all types of these tumors. Benign MPT had a lower Ki-67 than did borderline malignant MPT (4 versus 28). Malignant MPT had a greater than 8-fold higher Ki-67 activity than did benign tumors (35 versus 4). Intracyto-plasmatic c-kit expression was associated with a pathological diagnosis of malignant MPT, correlating with increasing grade (p < 0.05). In hypercellular stroma of borderline malignant and especially malignant forms of MPT, high activity of ER in mast cells was confirmed. Oncoprotein Her-2 activity, mostly in epithelial components, correlated with the degree of malignant progression of MPT (p < 0.05). CONCLUSION: Besides the well-known malignant features additional parameters have been found to be high Ki-67 and c-kit stromal expressions, and weak LAP activity in the epithelial part of malignant MPT, as well as mast cells with a high expression of ER.

[Granular-cell tumor: a rare variant of mammary tumor].

BACKGROUND: Granular cell tumor (GCT) is a rare variant of mammary tumor beset with diagnostic dilemmas that may be resolved by using numerous, very complex, enzymohistochemical and immunohistochemical methods. CASE REPORTS: We reported three female patients 16, 21 and 65 years old, operated on for mammary tumor at the Surgical Clinic of the School of Medicine in Nis, over the period of thirty years, 1977 to 2007. During this period 14.022 mammary tumors were diagnosed, including these three cases. These tumors had benign characteristics, without associated tumors in other localizations. A typical histological feature of GCT was a granular cytoplasm in large ovoid cells, organized like nests or like a trabecular arrangement. The tumors were analyzed by sets of histochemical, enzymohistochemical, immunohistochemical methods as well as ultrastructural examination. Protein, S-100 neuron-specific enolase and vimentin expressed a diffuse and intensive immunohistochemical activity, while expression of estrogen and progesterone receptors, as well as HER-2 oncoprotein was negative. The ultrastructural analysis confirmed that the tumor cells were enriched by lysosomes and consequential disorganization of cytoplasm. CONCLUSION: The reported enzymo- and immunohistochemical combined methods provide a precise diagnosis and confirm the GCT's neural origin, which has been disputed for years.

The effect of calcium channel blocker verapamil on gentamicin nephrotoxicity in rats.

Aminoglycoside antibiotics are obligated nephrotoxins and inevitably cause renal failure during prolonged use. Experimental models of gentamicin-induced nephrotoxicity have shown histopathological, ultrastructural and functional alteration with blood urea nitrogen and serum creatinine increase leading to acute renal insufficiency (ARI). The aim of our study was to emphasize effects of verapamil, a calcium channel blocker, on gentamicin-induced ARI in rats. Experiments were done on 50 male Wistar rats (250-300 g) divided in three experimental groups. G-group animals (20 rats) were treated daily with gentamicin in dose of 100 mg/kg during 8 days. GV-group animals (20 rats) were treated daily with verapamil in dose of 3 mg/kg and the same dose of gentamicin as in G-group during 8 days. The control group (10 rats) received 1 ml/day saline intraperitoneally. Histological examinations were done using hematoxylin and eosin, periodic acid Schiff and methenamine silver staining methods. Morphometric parameters included measurement of glomerular area, major and minor axis, perimeter, diameter, roundness, and mean optical density. Biochemical analyses were used to determine concentrations of blood urea, serum creatinine, sodium and potassium. In G-group rats' glomerular basement membrane was diffusely and unequally thickened with polymorphonuclear neutrophils infiltration, while coagulation-type necrosis and vacuolization of cytoplasm of proximal tubules epithelial cells were observed. In GV-group rats' glomeruli were slightly enlarged with thickened basement membrane in some segments but without coagulation-type necrosis. Morphometric analyses showed statistically significant differences between the G-group and control group of animals in glomerular size, mean optical density and average roundness (p<0,05). On the other hand, morphometric analyses between GV-group and control group animals did not show statistically significant differences in any of parameters measured. Blood urea and serum creatinine concentration in G-group were significantly elevated in comparison with GV-group (p<0,05) but sodium and potassium levels in G-group were decreased compared to GV-group without statistical significance. Our results show that verapamil modify some of morphological and functional kidney alterations induced by gentamicin.