Perfluoro-octanoic acid impairs sperm motility through the alteration of plasma membrane.

CONTEXT: Perfluoroalkyl-substances (PFAS) are chemical additives considered harmful for humans. We recently showed that accumulation of perfluoro-octanoic acid (PFOA) in human semen of exposed subjects was associated with altered motility parameters of sperm cells, suggesting direct toxicity. OBJECTIVES: To determine whether direct exposure of human spermatozoa to PFOA was associated to impairment of cell function. PATIENTS AND METHODS: Spermatozoa isolated from semen samples of ten normozoospermic healthy donors were exposed up to 2 h to PFOA, at concentrations from 0.1 to 10 ng/mL. Viability and motility parameters were evaluated by Sperm Class Analyser. Cell respiratory function was assessed by both mitochondrial probe JC-1 and respiratory control ratio (RCR) determination. Sperm accumulation of PFOA was quantified by liquid chromatography-mass spectrometry. Expression of organic ion-transporters OATP1 and SLCO1B2 was assessed by immunofluorescence and respective role in PFOA accumulation was evaluated by either blockade with probenecid or membrane scavenging through ß-cyclodextrin (ß-CD). Plasma membrane fluidity and electrochemical potential (ΔΨp) were evaluated, respectively, with Merocyanine-540 and Di-3-ANEPPDHQ fluorescent probes. RESULTS: Compared to untreated controls, a threefold increase of the percentage of non-motile sperms was observed after 2 h of exposure to PFOA regardless of the concentration of PFOA, whilst RCR was significantly reduced. Only scavenging with ß-CD was effective in reducing PFOA accumulation, suggesting membrane involvement. Altered membrane fluidity, reduced ΔΨp and sperm motility loss associated with exposure to PFOA were reverted by ß-CD treatment. CONCLUSION: PFOA alters human sperm motility through plasma-membrane disruption, an effect recovered by incubation with ß-CD.

Bioassay-guided isolation of the antinociceptive compounds motiol and beta-sitosterol from Scorzonera latifolia root extract.

Scorzonera latifolia (Asteraceae) is a plant widely distributed in Central and East Anatolia. A mastic, named yaki sakizi, is prepared from the latex and roots of S. latifolia and similar species. This latex is used in Turkish folk medicine for its analgesic activity, as anthelmintic and against infertility. The aim of this study was to isolate the compounds responsible for the antinociceptive activity of S. latifolia using bioassay-guided fractionation. The methanolic extract of the S. latifolia roots was prepared and subjected to chromatographic purification. Isolated active compounds were identified by means of MS and NMR techniques. Writhing and tail-flick tests were used to determine antinociceptive activity. Motiol and beta-sitosterol were isolated as compounds with promising antinociceptive activity. It is suggested that antinociceptive activity of the plant extract is probably caused by the synergic interaction of the isolated compounds.

Immunomodulation mediated by a herbal syrup containing a standardized Echinacea root extract: a pilot study in healthy human subjects on cytokine gene expression.

In this study, the immunomodulatory effect of a triply standardized Echinacea angustifolia root extract (Polinacea(®)) was evaluated in 10 healthy subjects. Ten ml of syrup containing one hundred mg of extract (corresponding to 4.7 mg of Echinacoside and 8.0mg of a high molecular weight-20,000 Da- polysaccharide) were administered as a herbal syrup once a day for one month. The immunomodulatory effect was evaluated before and after herbal syrup administration evaluating the expression levels of the cytokines IL-2, IL-8, IL-6 and TNF-α. Cytokine expression was studied in lympho-monocytes and in plasma samples measuring the mRNA and protein levels, respectively. The results were analysed by ANOVA and non-parametric Friedman rank sum tests; when possible it was adopted a pair-wise comparisons at different post-treatment times, using the paired t-tests with Holm correction. The correlation between the variations of cytokine plasma levels and the respective mRNA was carried out using a linear regression model. In lympho-monocytes our data indicate the up-regulation of the mRNA levels of IL-2 and IL-8 and the down regulation of the mRNA levels of the pro-inflammatory cytokines TNF-α and IL6. The differential regulation was maximal after 14 days of treatment. IL-2 up-regulation and IL-6 down-regulation were also confirmed at the protein level in plasma. Finally, the up-regulation of the mRNA of IL-2/IL-8 and the down-regulation of IL-6 positively correlated with the protein levels detected in the plasma. In conclusion, this pilot study suggests a relevant role for the standardized Echinacea angustifolia root extract in the control of cytokine expression. This first demonstration of the immuno-modulating activity of Echinacea angustifolia root extract in the healthy subject, supports at least in part the common use of such products as health promoting supplement.

A new triterpene from Scorzonera latifolia (Fisch. and Mey.) DC.

A novel triterpene 1 (3-ß-hydroxy-fern-7-en-6-one-acetate) together with four known compounds, urs-12-en-11-one-3-acetyl (2), 3-ß-hydroxy-fern-8-en-7-one-acetate (3), olean-12-en-11-one-3-acetyl (4) and leucodin (5) were obtained from the S. latifolia roots. All compounds were isolated from the n-hexane extract of S. latifolia roots using several chromatographic techniques. The structure of the isolated compounds was elucidated on the basis of (1)H-NMR, (13)C-NMR and 2D NMR data (HMBC, HMQC, COSY, TOCSY, NOESY, DEPT) as well as GC EITOF-HRMS.

Effect of sugar cane policosanol on lipid profile in primary hypercholesterolemia.

Policosanol, a mixture of long-chain aliphatic primary alcohols, is used as a cholesterol-lowering supplement. The effectiveness of policosanol is still questionable. To determine the lipoprotein-lowering effects of Cuban sugar cane-derived policosanol a double-blind, randomized, placebo controlled trial was performed. Sixty-eight primary hypercholesterolemic subjects were enrolled and randomly assigned to the treatment or to the control group. The first group received sugar cane policosanol 20 mg daily for 8 weeks, while the control group was treated with placebo. All subjects followed a normocaloric diet. The content of policosanol in the supplement tablets was assessed by gas chromatography. A total of 32 subjects in the policosanol group and 31 subjects in the control group completed the study. Body mass index, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride plasma levels did not change significantly in either group. In conclusion, sugar cane policosanol at doses of 20 mg daily showed no lipid lowering effects in subjects with primary hypercholesterolemia.

Analgesic and antiinflammatory activity of Cyclamen repandum S. et S.

According to folk medicine some species belonging to the genus Cyclamen were used for their biological activities. Early investigation of the different species of the genus resulted in the isolation of triterpenic saponins. No phytochemical and biological data are available on C. repandum. As part of a series of phytochemical investigations for bioactive compounds from medicinal plants, Cyclamen repandum S. et S. was investigated. The present study sought to find the antiinflammatory and antinociceptive activities of C. repandum tubers in rats and mice. A preliminary screening was conducted with three different extracts in the tests used, particularly the paw edema and the writhing tests. Subsequently some saponins isolated from the ME extract, the more effective one, have been identified. This paper also describes the results of fractionation and bioassay guided chemical studies. Chemical investigation of the active extract afforded the isolation and characterization of six triterpenic saponins. The possible antiinflammatory and analgesic properties were investigated as the saponin content of the fractions allows to speculate on such aspect.

In vitro estrogenic activity of Achillea millefolium L.

Isolation and biological characterization of pure compounds was used to identify and characterize estrogenic activity and estrogen receptors (ER) preference in chemical components of Achillea millefolium. This medicinal plant is used in folk medicine as an emmenagogue. In vitro assay, based on recombinant MCF-7 cells, showed estrogenic activity in a crude extract of the aerial parts of A. millefolium. After fractionation of the crude extract with increasing polar solvents, estrogenic activity was found in the methanol/water fraction. Nine compounds were isolated and characterized by HR-MS spectra and 1D- and 2D-NMR techniques. In particular, dihydrodehydrodiconiferyl alcohol 9-O-beta-D-glucopyranoside - a glycosyl-neolignan - was isolated for the first time from the genus Achillea in addition to six flavone derivatives, apigenin, apigenin-7-O-beta-D-glucopyranoside, luteolin, luteolin-7-O-beta-D-glucopyranoside, luteolin-4'-O-beta-D-glucopyranoside, rutin, and two caffeic acid derivatives, 3,5-dicaffeoylquinic acid and chlorogenic acid. Apigenin and luteolin, the most important estrogenic compounds among those tested, were studied for their ability to activate alpha or beta estrogen receptors (ERalpha, ERbeta) using transiently transfected cells. Our results suggest that isolation and biological characterization of estrogenic compounds in traditionally used medicinal plants could be a first step in better assessing further (e.g. in vivo) tests of nutraceutical and pharmacological strategies based on phytoestrogens.

Cytotoxic constituents from Anagyris foetida leaves.

Anagyris foetida extracts were investigated in preliminary cytotoxic tests against two tumour cell lines. Chromatographic separations on active extracts led to the isolation of two alkaloids, anagyrine (1) and baptifoline (2), as well of isorhamnetin (3) and syringin 4-O-beta-D-glucopyranoside (4). In vitro cytotoxicity of compounds 1-3 was also evaluated.

Topical anti-inflammatory activity of Solenostemma argel leaves.

The topical anti-inflammatory activity of Solenostemma argel Hayne leaves was evaluated using the Croton oil ear test in mice. A bioassay-guided fractionation procedure led to a highly active chloroform extract: at 300 microg/cm(2), it induced 73% oedema reduction, while the reference drug indomethacin (100 microg/cm(2)) induced 56% reduction. The extract contained a new pregnene glycoside (1, solenoside A) and the known 14 beta,15 alpha-dihydroxy-Delta(4)pregnene-3,20 dione (2), kaempferol-3-O-glucoside (3) and kaempferol-3-O-rutinoside (4). Their structures were determined by ID and 2D NMR experiments as well as HRMS. Compounds 2 and 4 showed anti-inflammatory activity comparable to that of indomethacin.

Anti-inflammatory, anti-nociceptive and antioxidant activities of Balanites aegyptiaca (L.) Delile.

The anti-inflammatory and anti-nociceptive activities of methanol (ME), butanol (BE) extracts and of two new saponins isolated from Balanites aegyptiaca bark were evaluated. The study was carried out in vivo and in vitro. The samples, extracts and pure substances, were intra-gastrically administered to animals. Two different animal models, the carrageenin-induced edema, in the rat, and acetic acid-induced writhing test in mice, were adopted. Moreover, the antioxidant power of extracts, fractions and individual constituents from Balanites aegyptiaca has been evaluated in vitro, using a method based on the Briggs-Rauscher (BR) oscillating reaction. Results obtained demonstrate that both ME or BE have a significant effect at the highest dose on the number of abdominal writhes induced by acetic acid, with a 38 and 54% inhibition respectively, but no significant difference was observed for extracts at the lowest dose and for the pure compounds compared with control animals. The same extracts exhibit a significant reduction on the rat paw edema. The inhibition produced by ME is about the same (28+/-3% lowest dose, 32+/-3% highest dose) after administration. A more evident effect is obtained by BE (41+/-3% and 68+/-6% respectively) and single saponins B1 and B2 (62+/-5% and 59+/-6% respectively) after oral administration. The antioxidant activity obtained seems to be in good accordance with the pharmacological results. The histological sections of rat paw confirm the antiflogistic activity of the plant extracts.

6-Aminoquinolones: photostability, cellular distribution and phototoxicity.

Three selected aminoquinolones endowed with a potent antibacterial (compounds 1 and 2) and antiviral activity (compound 3) have been evaluated for their phototoxic properties in vitro. Photostability studies of these compounds indicate that compound 3 is photostable whereas compound 1 and in particular, compound 2 are rapidly photodegraded upon UVA irradiation, yielding a toxic photoproduct. Intracellular localization of these compounds has been evaluated by means of fluorescence microscopy using tetramethylrhodamine methyl ester and acridine orange, which are specific fluorescent probes for mitochondria and lysosomes, respectively. No co-staining was observed with lysosomal stain for all the test compounds. On the contrary compound 3 was found to be specifically incorporated in mitochondria. The compounds exhibited remarkable phototoxicity in two cell culture lines: human promyelocytic leukaemia (HL-60) and human fibrosarcoma (HT-1080). The quinolone-induced photodamage was also evaluated measuring the photosensitizing cross-linking in erythrocyte ghost membranes, the strand breaks activity and oxidative damage on plasmid DNA. The results show that these derivatives are able to photoinduce crosslink of erythrocytes spectrin, whereas do not significantly photocleavage DNA directly, but single strand breaks were observed after treatment of photosensitized DNA with two base excision repair enzymes, Fpg and Endo III respectively.

[Hyperparathyroidism in patients with respiratory decompensation]. / Iperparatiroidismo in pazienti in scompenso respiratorio.

Eighteen patients aged between 55 and 65 and affected by respiratory insufficient were included in the study. Serum levels of PTH, total calcium, Ca++ and phosphorus were measured. Special attention was focused on PTH concentrations with in some case provedo to be above normal. Two hypotheses of pathogenesis are put forward. The first suggests that reduced Ca++ levels are responsible for anomalous PTH serum concentrations, whereas the second suggests that this role may be played by hypoxia. The latter would indirectly or directly affect parathyroids, either increasing PTH hyperincretion or slowing down the metabolism of the hormone by kidneys and liver which are in turn negatively affected by low blood oxygen tension.

[Tumor markers in colorectal neoplasm pathology: our experience of the diagnostic and prognostic importance of CEA antigen]. / I markers tumorali nella patologia neoplastica del colon-retto: nostra esperienza sul significato diagnostico e prognostico dell'antigene CEA.

It is well known that the presence of a tumour may be accompanied by production and release into the serum of a substantial number of protein and/or hormonal substances. Only relatively recently, however, have investigators begun to identify which of these supposed markers may actually be clinically useful. A picture is thus gradually emerging of a number of markers (in actual fact only very few) which are already clinically useful, as well as others which are clinically promising and numerous markers which require more thorough clinical evaluation. Prominent among those already in clinical use is undoubtedly CEA. Carcinoembryonic antigen (CEA) is a glycoprotein isolated for the first time by Gold and Friedman in 1965. Very recently, sensitive radioimmune and immuno-enzymatic assays have made it possible to determine serum levels lower than 3 ng/ml. Though high plasma levels of CEA do not indicate the presence of a tumour with certainty, very high levels are, however, to some extent indicative of its existence. In view of the fact that CEA possesses neither the sensitivity nor the specificity to be able to diagnose the presence or otherwise of a tumour, its use is generally recommended when formulating a prognosis or for monitoring surgical and/or medical therapy (chemotherapy and radiotherapy) in asymptomatic patients. We therefore carried out tests in 357 patients on file as having undergone surgery for neoplasms of the colon-rectum-anus, monitored in our colostomy outpatients' department, which was started up in 1980. 188 of these patients had already been submitted to CEA assay prior to surgery. The data are analyzed in relation to the site, stage and grade of differentiation of the neoplasm, the supposed radicality of the operation and the period of follow-up in these patients.

[Medullary carcinoma of the thyroid. Observation of a case]. / Il carcinoma midollare della tiroide. Osservazione di un caso.

A case of medullary carcinoma of the thyroid is reported, particular attention being paid to preoperative diagnostic problems. The importance of cytological examination carried out with fine needle acupuncture is also stressed. The anatomopathological and clinical features are outlined, special mention being made of the neoplasia's frequent hormonal secretion and its possible familial nature.

[Solitary congenital cyst of the liver. Clinical and therapeutic considerations on 2 cases]. / Cisti congenita solitaria del fegato. Considerazioni clinico-terapeutiche su 2 casi.

A brief overview of the aetiopathogenetic aspects of non-parasitic cyst of the liver is followed by the presentation of two personal cases. Their diagnostic, anatomopathological and therapeutic features are examined, and reference is made to the treatment employed, namely partial cystectomy and derivation on an excluded loop à la Roux. It is suggested that the type of surgery employed will depend in each case on the relation of the cyst to the liver parenchyma (superficial or deep sites) and neighbouring organs. Benign forms should be treated in a not excessively destructive manner.

[Experiments with splenohepatoplasty in the rat]. / Esperimenti di splenoepatoplastica nel ratto

The physiopathological premises for various methods of hepatic revascularisation are described and results wiht splenohepatoplasty in 40 rats are presented. This operation involves insertion of the upper pole of decapsulated spleen in the left hepatic lobe to create a new parenchymal supply and regenerate the liver cell. The clinical, functional and anatomopathological data are discussed. The experimental value of the procedure is asserted, though it is felt that no sure demonstration can be given of the establishment of newly formed vascular connections between the two organs.