Expression profiling of extramedullary acute myeloid leukemia suggests involvement of epithelial-mesenchymal transition pathways.

Extramedullary (EM) colonization is a rare complication of acute myeloid leukemia (AML), occurring in about 10% of patients, but the processes underlying tissue invasion are not entirely characterized. Through the application of RNAseq technology, we examined the transcriptome profile of 13 AMLs, 9 of whom presented an EM localization. Our analysis revealed significant deregulation within the extracellular matrix (ECM)-receptor interaction and focal-adhesion pathways, specifically in the EM sites. The transcription factor TWIST1, which is known to impact on cancer invasion by dysregulating epithelial-mesenchymal-transition (EMT) processes, was significantly upregulated in EM-AML. To test the functional impact of TWIST1 overexpression, we treated OCI-AML3s with TWIST1-siRNA or metformin, a drug known to inhibit tumor progression in cancer models. After 48 h, we showed downregulation of TWIST1, and of the EMT-related genes FN1 and SNAI2. This was associated with significant impairment of migration and invasion processes by Boyden chamber assays. Our study shed light on the molecular mechanisms associated with EM tissue invasion in AML, and on the ability of metformin to interfere with key players of this process. TWIST1 may configure as candidate marker of EM-AML progression, and inhibition of EMT-pathways may represent an innovative therapeutic intervention to prevent or treat this complication.

Comparison between azacitidine and decitabine as front-line therapy in elderly acute myeloid leukemia patients not eligible for intensive chemotherapy.

We compared the efficacy of azacitidine (AZA) and decitabine (DEC) in elderly patients with untreated AML, diagnosed according to WHO criteria. In the two groups, we evaluated complete remission (CR), overall survival (OS) and disease free survival (DFS). The AZA and DEC groups included 139 and 186 patients, respectively. To minimize the effects of treatment selection bias, adjustments were made using the propensity-score matching method, which yielded 136 patient pairs. In the AZA and DEC cohort, median age was 75 years in both, (IQR, 71-78 and 71-77), median WBCc at treatment onset 2.5 × 109/L (IQR, 1.6-5.8) and 2.9 × 109/L (IQR, 1.5-8.1), median bone marrow (BM) blast count 30% (IQR, 24-41%) and 49% (IQR, 30-67%), 59 (43%) and 63 (46%) patients had a secondary AML, respectively. Karyotype was evaluable in 115 and 120 patients: 80 (59%) and 87 (64%) had intermediate-risk, 35 (26%) and 33 (24%) an adverse risk karyotype, respectively. Median number of cycles delivered was 6 (IQR, 3.0-11.0) and 4 (IQR, 2.0-9.0), CR rate was 24% vs 29%, median OS and 2-year OS rates 11.3 (95% CI 9.5-13.8) vs 12.0 (95% CI 7.1-16.5) months and 20% vs 24%, respectively. No differences in CR and OS were found within the following subgroup: intermediate- and adverse-risk cytogenetic, frequency of WBCc at treatment ≥ 5 × 10^9 L and < 5 × 10^9/L, de novo and secondary AML, BM blast count < and ≥ 30%. Median DFS for AZA and DEC treated patients was 9.2 vs 12 months, respectively. Our analysis indicates similar outcomes with AZA compared to DEC.

The emerging role of measurable residual disease detection in AML in morphologic remission.

Despite the increasing knowledge of the genomic landscape of acute myeloid leukemia (AML), prediction merely based on genetics fails to anticipate outcome, presumably due to the heterogeneous composition of the leukemic clone determining complex interactions between different genetic abnormalities. Therefore, the introduction of a post-treatment biomarker exploring the quality of response to therapy such as assessment of measurable (previously minimal) residual disease (MRD) may lead to refinements of the prognostic assessment in AML. In this view, the European LeukemiaNet has recently endorsed the achievement of a MRD negative morphologic complete remission as a purpose the treatment. Techniques like multiparametric flow cytometry and reverse transcriptase-quantitative polymerase chain reaction have reached a level of sensitivity and specificity that make them ready for introduction in clinical practice. In the present review, we will give an update on the efforts in harmonization and/or standardization of MRD assessment in AML, focusing on the newest acquisitions in the clinical applications of MRD, and considering issues like relationship of MRD with leukemic stem cells or MRD assessment in peripheral blood.

Low-level laser therapy in the treatment of muscle-skelet pain in patients affected by temporo-mandibular disorders.

BACKGROUND: The purpose of the study is to evaluate the effectiveness of Low-Level Laser Therapy in reducing joint and muscle pain in patients with acute and chronic temporomandibular dysfunction. MATERIALS AND METHODS: The study was conducted on a sample of 180 patients. The sample was divided into two groups according to the time of onset of the disease: acute TMD (<6 months) and chronic TMD (> 6 months). The treatment for all patients provided for the irradiation with Diode Laser Wiser Doctor Smile with tip plane wave at wavelength of 830 nm, continuous beam to 40nW diameter and radius of 6 mm. The irradiated areas were the joint area, temporal, masseter and pterygoid. The irradiation time for each zone was 60s.The protocol adopted consisted of two weekly treatment for six weeks. Pain assessment was performed using the Visual Analog Scale (VAS), in which different scores (s) depending accused of pain by the patient: s0 no pain, s1-3 mild pain, s4-6 moderate pain, s7-9 severe pain and s10 excessive pain. The pain monitoring was performed before treatment, after 15 days and after one month. RESULTS: The sample included 80 patients with acute TMD and 100 with chronic TMD. The sample belonging to acute TMD group before treatment, was distributed as follows: 0% in s0; 12,5% in s1-3; 31.3% in s4-6; 53.6% in s7-9 and 2.5% in s10. After 15 days the distribution was was as follows: 6.25% in s0; 47.5% in s1-3; 20% in s4-6; 26.3% in s7-9 and 0% in s10. After 30 days the sample was well distributed: 35% in s0; 45% in 1-3; 10% in s4-6; 10% in s7-9 and 0% in s10. The sample belonging to the chronic TMD group, at time zero, was as follows: 0% at s0; 48% in S1-3; 35% in s4-6; 15% in s7-9 and 2% in s10. After 15 days the distribution was: 29% in s0; 28% in S1-3; 33% in s4-6; 10% in s7-9 and 0% in s10. After 30 days the sample was well distributed: 45% in s0; 36% in S1-3; 15% in s4-6; 4% in s7-9 and 0% in s10. CONCLUSIONS: The Low-Level-Laser-Therapy is a valuable tool that can significantly decrease the perception of pain in patients with temporomandibular joint dysfunction, acute and chronic.

Polysonographical evaluation in a case of moderate osas treated with mandibular advancement device.

BACKGROUND: The Obstructive Sleep Apnea Syndrome (OSAS) is a clinical picture characterized by partial or complete obstruction of the upper airway during sleep, associated with a reduction of oxygen saturation in the blood.The most common symptoms are: apnea sleep, snoring, headache, sleepiness, reduced concentration and memory, irritability, increased blood pressure and dry mouth. MATERIALS AND METHODS: It was examinated a not-smoker man of 54 years that suffers of roncophaty. He did physical examination, rhinoscopyexam, faringoscopy, rhinofibrolaringoscopy, gnathological evaluation and polysomnographic examination performed with multichannel polygraphy (VitalNight). From the performed examinations, it has been diagnosed a moderate form of obstructive apnea sleep syndrome. He was treated with a mandibular advancement device and it was repeated the polysomnographic exam. RESULTS: Comparing the results of the polysonographic examination performed before and after the treatment, the patient's clinical picture clearly improved. It has shown a clear reduction of obstructive apneas, hypopneas and snoring. CONCLUSIONS: The use of a mandibular advancement device is certainly a valuable aid in the treatment of moderate type OSAS. The quality of diurnal life is also improved as shown with Sleepness Epworth Scale.

Phytochemistry, micromorphology and bioactivities of Ajuga chamaepitys (L.) Schreb. (Lamiaceae, Ajugoideae): Two new harpagide derivatives and an unusual iridoid glycosides pattern.

Ajuga chamaepitys (L.) Schreb, well-known as Camaepitium or Ground Pine, is an annual herb typical of the Mediterranean area accounting several uses in the traditional medicine. In this work we have, analyzed the plant iridoid fraction together with the essential oil composition and study of the plant indumentum. Finally, we assayed the polar extracts and essential oil obtained from the aerial parts for antioxidant activity and cytotoxicity on tumor cells. The analysis of the monoterpene glycosides allowed us to isolate from roots and aerial parts and to structurally elucidate by NMR and MS the following compounds: ajugoside (1), reptoside (2), 8-O-acetylharpagide (3), harpagide (4), 5-O-ß-d-glucopyranosyl-harpagide (5), asperulosidic acid (6), deacetyl asperulosidic acid (7) and 5-O-ß-d-glucopyranosyl-8-O-acetylharpagide (8), among which 5 and 8 were two new natural products. Chemotaxomic relevance of these constituents was discussed. The chemical analysis of A. chamaepitys essential oil by GC-FID and GC-MS showed ethyl linoleate (13.7%), germacrene D (13.4%), kaurene (8.4%), ß-pinene (6.8%), and (E)-phytol (5.3%) as the major volatile components. The micromorphological and histochemical study showed that iridoids and essential oil are mainly produced in the type III capitates and peltate trichomes of leaves and flowers. Biological evaluations of A. chamaepitys polar extracts and essential oil showed that the former were more potent as radical scavengers than the latter. MTT assay revealed that essential oil and ethanolic extracts were moderately cytotoxic on tumor cells with IC50 of 36.88 and 59.24µg/mL on MDA-MB 231 cell line, respectively, and IC50 of 60.48 and 64.12µg/mL on HCT116, respectively.

Venetoclax: Bcl-2 inhibition for the treatment of chronic lymphocytic leukemia.

Venetoclax (ABT-199) is a small-molecule selective oral inhibitor of the antiapoptotic protein Bcl-2 that promotes programmed cell death of chronic lymphocytic leukemia (CLL) cells regulating the release of proapoptotic factors, such as Smac/Diablo, apoptosis-inducing factor (AIF) and cytochrome c. In April 2016, the U.S. Food and Drug Administration (FDA) granted accelerated approval to venetoclax for patients diagnosed with CLL with 17p deletion, as detected by an FDA-approved test, who have received at least one prior therapy. This review will focus on the mechanism of action, preclinical studies and clinical development of venetoclax both as a monotherapy and in combination with other drugs for CLL in the current milieu of therapy dominated by novel tyrosine kinase inhibitors such as ibrutinib and idelalisib.

Statistical analysis of the frequency distribution of signs and symptoms of patients with temporomandibular disorders.

The purpose of this text is to evaluate the distribution by sex and age of TMD patients and the frequency of pain on palpation of the lateral pterygoid muscle, pterygoid internal, sternocleidomastoid, masseter, temporal. The purpose is also to assess the existence of direct correlations between the articulating click and lateral deviation, the headache and tenderness of the temporal, the back pain and tenderness of the sternocleidomastoid, the postural disorders and difficulty finding the exact mating closing, the disorders related to previous dental procedures (large conservative reconstructions, rehabilitations fixed dentures and dental extractions) and tenderness to the temporal and masseter. 623 medical records, including 451 males and 175 females were randomly selected. It was statistically calculated the F/M ratio, age distribution and frequency of muscle tenderness to palpation through the relationship of positive feedback on the total sample multiplied by one hundred. The correlation between various disorders was calculated by estimating the Mantel-Haenszel common odds ratio. For each result it has been finally calculated the asymptotic level of significance.

Unusual molecular pattern in Ajugoideae subfamily: the case of Ajuga genevensis L. from Dolomites.

We analysed the ethanolic extract from Ajuga genevensis L. (Lamiaceae) growing in Dolomites, part of Italian Alps. Three new compounds for this species were identified: rosmarinic acid (1), oleanolic acid (2) and maslinic acid (3), representative of two different classes of chemical compounds (phenylpropanoids and pentacyclic triterpenes). A. genevensis resulted to be a valuable source of these compounds endowed with interesting biological activities (i.e. antioxidant, neuroprotective, anti-inflammatory, antiproliferative). The recognition of compounds (1), (2) and (3) may also confirm the ethnomedicinal uses of this plant. From a chemotaxonomical point of view, it is worth noting that iridoids were not evidenced in this accession. Iridoids are considered chemotaxonomic marker in Lamiales, and, in contrast with a previous study on this species, the presence of aucubin was not confirmed. In addition, the presence of large amounts of rosmarinic acid (1) was unexpected for a species that does not belong to subfamily Nepetoideae.

Secondary metabolites from Scrophularia canina L.

A re-examination of Scrophularia canina L. confirmed the presence of iridoid glucosides considered as chemotaxonomic markers for the Scrophulariaceae family, like aucubin, harpagide and 8-O-acetylharpagide, besides the further presence of 8-epiloganic acid, which is, indeed, considered the biogenetic precursor of iridoids normally found in Scrophulariaceae, and was recognised here for the first time in the studied species. Also verbascoside and (E)-phytol were evidenced for the first time in S. canina. The former compound is an almost ubiquitous glycosidic phenyl-ethanoid, which attains systematic importance when in co-occurrence with iridoids, and its taxonomical implications were discussed. The latter compound, even though it is omnipresent, is interestingly endowed with several biological activities, which may give an additional reason for the traditional uses of this plant.

Scenedesmus dimorphus (Turpin) Kützing growth with digestate from biogas plant in outdoor bag photobioreactors.

Digestate coming from an Anaerobic Digestion unit in a Biogas Plant, feeded on cow manure and vegetable waste from markets, has been used. About 8-35 L polyethylene transparent bags have been employed as cultivation container, outdoor. Different aliquots of digestate, alone or mixed with commercial liquid fertiliser, were employed to cultivate in batch Scenedesus dimorphus, a freshwater green microalga, in the ENEA facilities of Casaccia Research Center, near Rome, Italy. The cultivation period was June-July 2013. The average daily yields of dry microalgae biomass varied from 20 mg/L/d to 60 mg/L/d, mean 38.2 mg/L/d. Final dry biomass concentration varied from 0.18 to 1.29 g/L, mean 0.55 g/L. S. dimorphus proved to be very efficient in removing N and P from the culture medium. Another fact emerged from these trials is that S. dimorphus inner composition resulted to be variable in response to the tested different culture conditions.

Reassessment of Melittis melissophyllum L. subsp. melissophyllum iridoidic fraction.

The analysis of the polar fraction of Melittis melissophyllum L. subsp. melissophyllum led to the identification of several iridoid glycosides: monomelittoside (1), melittoside (2), harpagide (3), acetyl-harpagide (4) and ajugoside (5). Compounds 3 and 4 are considered marker compounds for the genus and, as well as compounds 1, 2 and 5, were already evidenced in a previous study on the nominal species. It was noteworthy of the presence of allobetonicoside (6) which was never reported for this genus. The isolation of 6 is very relevant because of its allose residue on the structure. Allose has been often found in the species of the subfamily Lamioideae even if it mostly regarded flavonoids considered of chemotaxonomical relevance for some correlated genera of Lamiaceae. Same as allosyl-glycosidic flavonoids, the presence of allosyl-glycosidic iridoids may also be an additional chemosystematic evidence of botanical relationships among Lamiaceae species and genera.

Minimal residual disease negativity in elderly patients with acute myeloid leukemia may indicate different postremission strategies than in younger patients.

In the present analysis, we evaluated whether in elderly acute myeloid leukemia (AML) patients (>60 years), minimal residual disease (MRD) assessed by flow cytometry may have a role in guiding choice of postremission strategies. We analyzed 149 young and 61 elderly adults who achieved morphological CR after induction course of EORTC/GIMEMA protocols. Elderly patients reached a postconsolidation MRD negative status less frequently than younger ones (11 vs 28 %, p = 0.009). MRD negativity resulted in a longer 5-year disease-free survival (DFS) both in elderly (57 vs 13 %, p = 0.0197) and in younger patients (56 vs 31 %, p = 0.0017). Accordingly, 5-year cumulative incidence of relapse (CIR) of both elderly (83 vs 42 %, p = 0.045) and younger patients (59 vs 24 % p = NS) who were MRD positive doubled that of MRD negative ones. Nevertheless, CIR of MRD negative elderly patients was twofold higher than that of younger MRD negative ones (42 vs 24 %, p = NS). In conclusion, elderly patients in whom chemotherapy yields a MRD negative CR have duration of DFS and rate of CIR significantly better than those who remain MRD positive. Nonetheless, the high CIR rate observed in the elderly suggests that MRD negativity might have different therapeutic implications in this population than in the younger counterpart.

Terpenoids of Linaria alpina (L.) Mill. from Dolomites, Italy.

This paper reports the first phytochemical analysis of Linaria alpina (L.) Mill., collected in Dolomites (Italy), a species characteristic of mountain environment. Besides aucubin (4), which is rare in the subgenus Antirrhineae of Plantaginaceae, mainly acidic compounds were found, i.e. oleanolic acid (1), ursolic acid (2) maslinic acid (3) and shikimic acid (5). The pentacyclic triterpenes of L. alpina resulted in relatively high content, whereas flavonoids resulted in low content.

Azacitidine frontline therapy for unfit acute myeloid leukemia patients: clinical use and outcome prediction.

Hypomethylating agents are able to prolong the overall survival of some patients diagnosed with acute myeloid leukemia. The aim of this study was to evaluate the clinical use of azacitidine as front-line therapy in unfit acute myeloid leukemia (AML) patients and to develop a clinical prediction model to identify which patients may benefit more from the drug. One hundred and ten untreated unfit AML patients received front-line azacitidine therapy in Spain, and response and survival were evaluated in them following European LeukemiaNet (ELN) guidelines. A clinical prediction rule was obtained from this population that was validated and refined in 261 patients treated in France, Austria and Italy. ELN response was achieved in 21.0% of the 371 patients (CI95% 17.0-25.5) and did not depend on bone marrow blast cell percentage. Median overall survival was 9.6 months (CI95% 8.5-10.8) and 40.6% of the patients were alive at 1 year (CI95% 35.5-45.7). European ALMA score (E-ALMA), based on performance status, white blood cell counts at azacitidine onset and cytogenetics, discriminated three risk groups with different survival and response rates. Azacitidine seems a reasonable therapeutic option for most unfit AML patients, i.e. those displaying a favorable or intermediate E-ALMA score.

Monoterpenoids glycosides content from two Mediterranean populations of Crucianella maritima L.

In this study, the iridoidic content of two accessions of Crucianella maritima L., one from Sardinia and the second from Latium, was examined and compared. From a qualitative point of view, the iridoidic pattern of the two samples was similar, since the same compounds (asperuloside, asperulosidic acid and deacetyl asperulosidic acid) were isolated. Asperuloside was the main compound in both accessions. Asperulosidic acid was the second compound in the accession from Sardinia, while the accession from Latium exhibited a similar amount of asperulosidic acid and deacetyl asperulosidic acid. These iridoids can be considered as chemotaxonomic markers for parts of the Rubiaceae family, in particular for the Rubioideae subfamily to which C. maritima belongs.

Non steroid treatment for eczema: results from a controlled and randomized study.

AIM: This study assesses the efficacy of a new non steroid anti-inflammatory product in comparison to Hydrocortisone Butyrate 0.1% Cream in healing eczematous dermatitis. METHODS: A bilateral controlled randomized pilot study was conducted in Italian adults affected by eczema with at least two symmetric lesions at baseline, respectively assigned to a non steroid cream or Hydrocortisone. The severity of lesions was judged through the Global Clinical Score (GCS) and the recovery was defined as a GSC equal to 0. The study investigated: 1) the differences in GCS between four points in time during therapy (baseline, four, eight, twelve weeks), according to medication received; 2) treatment efficacy. RESULTS: The study showed that time, treatment and interaction between treatment and time were associated with GCS; moreover, lesions treated with Hydrocortisone went better on the whole but the post-hoc analysis showed a significant clinical improving at each point in time only for the non steroid cream. At the end of the study, in the intention to treat analysis, lesions recovered in 76.1% and 40.3% patients treated with Hydrocortisone and with the non steroid cream respectively; in the per protocol population, recovery was achieved in 91.7% and 58.3% of cases. CONCLUSION: According to the results, the non steroid cream has been demonstrated effective in reducing the severity of eczema and may be used with continuing success in the long term treatment of the disease.

Phytochemical pattern of Gentiana species of Appennino in central Italy.

The molecular pattern of two Gentiana species, G. dinarica and G. lutea, present in a protected area of Appennino Centrale in Italy, was examined. Results were compared with literature data, examining the differences between the two species.