Improving sustainable hydrogen production from green waste: [FeFe]-hydrogenases quantitative gene expression RT-qPCR analysis in presence of autochthonous consortia.

BACKGROUND: Bio-hydrogen production via dark fermentation of low-value waste is a potent and simple mean of recovering energy, maximising the harvesting of reducing equivalents to produce the cleanest fuel amongst renewables. Following several position papers from companies and public bodies, the hydrogen economy is regaining interest, especially in combination with circular economy and the environmental benefits of short local supply chains, aiming at zero net emission of greenhouse gases (GHG). The biomasses attracting the largest interest are agricultural and urban green wastes (pruning of trees, collected leaves, grass clippings from public parks and boulevards), which are usually employed in compost production, with some concerns over the GHG emission during the process. Here, an alternative application of green wastes, low-value compost and intermediate products (partially composted but unsuitable for completing the process) is studied, pointing at the autochthonous microbial consortium as an already selected source of implementation for biomass degradation and hydrogen production. The biocatalysts investigated as mainly relevant for hydrogen production were the [FeFe]-hydrogenases expressed in Clostridia, given their very high turnover rates. RESULTS: Bio-hydrogen accumulation was related to the modulation of gene expression of multiple [FeFe]-hydrogenases from two strains (Clostridium beijerinckii AM2 and Clostridium tyrobutyricum AM6) isolated from the same waste. Reverse Transcriptase quantitative PCR (RT-qPCR) was applied over a period of 288 h and the RT-qPCR results showed that C. beijerinckii AM2 prevailed over C. tyrobutyricum AM6 and a high expression modulation of the 6 different [FeFe]-hydrogenase genes of C. beijerinckii in the first 23 h was observed, sustaining cumulative hydrogen production of 0.6 to 1.2 ml H2/g VS (volatile solids). These results are promising in terms of hydrogen yields, given that no pre-treatment was applied, and suggested a complex cellular regulation, linking the performance of dark fermentation with key functional genes involved in bio-H2 production in presence of the autochthonous consortium, with different roles, time, and mode of expression of the involved hydrogenases. CONCLUSIONS: An applicative outcome of the hydrogenases genes quantitative expression analysis can be foreseen in optimising (on the basis of the acquired functional data) hydrogen production from a nutrient-poor green waste and/or low added value compost, in a perspective of circular bioeconomy.

P450-based porous silicon biosensor for arachidonic acid detection.

A porous silicon biosensor based on P450 enzyme for arachidonic acid detection was developed. A new transduction method is presented with a simultaneous measurement of refractive index and fluorescence intensity changes when the analyte is binding to an enzyme on the porous silicon surface. A fluorophore bound to a cysteine residue in an allosteric position of the haem domain (BMP) of cytochrome P450 BM3 enhances its fluorescence intensity upon interaction with its substrate arachidonic acid, involved in diseases such as Alzheimer's, liver cancer and cellular inflammation processes. BMP has been anchored on porous silicon surface and the new transduction method has been successfully exploited to develop a biosensor for arachidonic acid, reaching a detection limit of 10 µM arachidonic acid in a dynamic range of 10-200 µM. Moreover, the change of the refractive index has been also monitored at the same time, displaying a higher detection limit of 30 µM. Preliminary test were also conducted in plasma proving the high specificity and selectivity of the sensor even in presence of interferents in the range of 50-100 µM. Here we suggest these two detection systems could be used simultaneously to increase the accuracy and the dynamic range of the sensor avoiding a false positive response.

First Report of Leaf Spot Caused by Pseudomonas cichorii on Coreopsis lanceolata in Italy.

Coreopsis lanceolata L. (Compositae), an ornamental species grown in parks and gardens, is very much appreciated for its long-lasting flowering period. In August of 2008, pot-grown plants with necrotic leaf lesions were observed in a commercial nursery located near Biella (northern Italy). Lesions were present, especially along the margin of basal leaves, and sometimes had a chlorotic halo. On infected leaves, dark brown necrosis developed. Leaf stalks were sometimes affected. In many cases, the leaves, especially those at collar level, were withered. Of 1,500 plants, 15% were infected by the disease. Microscopic examination did not reveal any fungal structures within the lesions. Small fragments of tissue from 30 affected leaves were macerated for 15 min in casein hydrolysate and 0.1-ml aliquots of the resulting suspension were spread onto Luria Bertani agar (LB) and potato dextrose agar (PDA). Plates were maintained at 22 ± 1°C for 48 h. No fungi were isolated from the leaf spots on LB or PDA. Colonies similar to those of Pseudomonas spp. were consistently isolated on LB. Colonies were fluorescent on King's medium B, levan negative, oxidase positive, potato soft rot negative, arginine dihydrolase negative, and tobacco hypersensitivity positive (LOPAT test). The bacterial colonies were identified as Pseudomonas cichorii (2). The internal transcribed spacer (ITS) region of rDNA was amplified using primers 27F and 1492R and sequenced (GenBank Accession No. FJ534557). BLAST analysis (1) of the 998-bp segment showed a 98% homology with the sequence of P. cichorii. The pathogenicity of one isolate was tested twice by growing the bacterium in nutrient broth shake cultures for 48 h at 20 ± 1°C. The suspension was centrifuged, the cell pellet resuspended in sterile water to a concentration of 107 CFU/ml, and 30 4-month-old healthy coreopsis plants were sprayed with the inoculum. The same number of plants was sprayed with sterile nutrient broth as a control. After inoculation, plants were covered with plastic bags for 48 h and placed in a growth chamber at 20 ± 1°C. Five days after inoculation, lesions similar to those seen in the field were observed on all plants inoculated with the bacterium, but not on the controls. Ten days later, 40% of the leaves were withered. Isolations were made from the lesion margins on LB and the resulting bacterial colonies were again identified as P. cichorii. The pathogen caused the same symptoms also on plants of Dendranthema frutescens (cv. Camilla), Chrysanthemum morifolium (cvs. Eleonora and Captiva), and an Osteospermum sp. (cv. Wild side) when artificially inoculated with the pathogen with the same methodology. The same bacterial leaf spot caused by P. cichorii was observed in 2005 in other nurseries in the same area on Phlox paniculata (3). To our knowledge, this is the first report of bacterial leaf spot caused by P. cichorii on C. lanceolata in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) H. Bergey et al. Bergey's Manual on Determinative Bacteriology. Williams and Wilkins, Baltimore, MD, 1994. (3) A. Garibaldi et al. Plant Dis. 89:912, 2005.

Report of Leaf Blight on Washington Lupine (Lupinus polyphyllus) Caused by Rhizoctonia solani AG 4 in Italy.

Lupinus polyphyllus (Leguminosae), Washington lupine, is a perennial herbaceous plant. In March 2008, in a campus greenhouse at the University of Torino, Grugliasco (northern Italy), a leaf blight was observed on 20% of potted 30-day-old plants. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Severely infected plants died. Plants were grown in a sphagnum peat/perlite/clay (70:20:10) substrate at temperatures between 18 and 25°C and relative humidity of 60 to 80%. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani (4) was consistently and readily recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 20 ± 1°C appeared light brown, rather compact, and exhibited radial growth. The isolates of R. solani successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (3). Pairings were also made with tester isolates AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 660-bp fragment showed 100% homology with the sequence of R. solani. The nucleotide sequence has been assigned GenBank Accession No. FJ486272. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old L. polyphyllus were grown in 10-liter containers (10 plants per container) on a steam disinfested sphagnum peat/perlite/clay (70:20:10) medium. Inoculum, consisting of an aqueous suspension of mycelium disks prepared from PDA cultures (5 g of mycelium per plant), was placed at the collar of plants. Plants inoculated with water and PDA fragments alone served as control treatments. Three replicates were used. Plants were maintained in a greenhouse at temperatures between 18 and 23°C. First symptoms, similar to those observed in the nursery, developed 10 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was repeated twice. The susceptibility of L. polyphyllus to R. solani was reported in Poland (2). This is, to our knowledge, the first report of leaf blight of L. polyphyllus caused by R. solani in Italy. The importance of the disease is at the moment limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) W. Blaszczak. Rocz. Nauk. Roln. Ser A 85:705, 1962. (3) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (4) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.

First Report of Leaf Blight on Fan Columbine (Aquilegia flabellata) Caused by Rhizoctonia solani AG 4 in Italy.

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips. It can also be grown for cut flower production. In April of 2008, in several nurseries located near Biella (northern Italy), a leaf blight was observed on 10 to 15% of potted 30-day-old plants grown on a sphagnum peat substrate at 15 to 20°C and relative humidity of 80 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded over several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and abscised. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and had radial growth. Sclerotia were not present. Isolates obtained from affected plants successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates of AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 648-bp fragment showed a 100% homology with the sequence of R. solani AG-4 AB000018. The nucleotide sequence has been assigned GenBank Accession No. FJ 534555. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Five plants of 30-day-old A. flabellata were grown in 3-liter pots. Inoculum consisting of an aqueous suspension of PDA and mycelium disks (5 g of mycelium + agar per plant) was placed at the collar of plants. Five plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a greenhouse at temperatures between 20 and 24°C. The first symptoms, similar to those observed in the nursery, developed 7 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. The presence of R. solani AG1-IB on A. flabellata has been reported in Japan (4), while in the United States, Rhizoctonia sp. is described on Aquilegia sp. (3). This is, to our knowledge, the first report of leaf blight of A. flabellata caused by R. solani in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) E. Imaizumi et al. J. Gen. Plant Pathol. 66:210, 2000.

First Report of Powdery Mildew Caused by Podosphaera xanthii on Calendula officinalis in Italy.

Calendula officinalis L. (Asteraceae) (pot marigold or English marigold) is an ornamental species grown in gardens and as potted plants for the production of cut flower. It was also used in ancient Greek, Roman, Arabic, and Indian cultures as a medicinal herb as well as a dye for fabrics, foods, and cosmetics. During the summer of 2007, severe outbreaks of a previously unknown powdery mildew were observed on plants in several gardens near Biella (northern Italy). Both surfaces of leaves of infected plants were covered with dense, white mycelia and conidia. As the disease progressed, infected leaves turned yellow and died. Mycelia and conidia also were observed on stems and flower calyxes. Conidia were hyaline, ellipsoid, born in short chains (four to six conidia per chain), and measured 27.0 to 32.1 (31.4) × 12.9 to 18.4 (18.2) µm. Conidiophores measured 49 to 77.3 (67.2) × 8 to 13.3 (10.8) µm and showed a foot cell measuring 44 to 59 (51.9) × 9.3 to 12.6 (11.3) µm followed by one shorter cell measuring 15.6 to 18.9 (17.6) × 10.4 to 13.6 (12.2) µm. Fibrosin bodies were present. Chasmothecia were spherical, amber colored, with a diameter of 89 to 100 (94.5) µm. Each chasmothecium contained one ascus with eight ascospores. On the basis of its morphology, the causal agent was determined to be a Podosphaera sp. (2). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 588 bp showed a 100% homology with the sequence of Podosphaera xanthii (2). The nucleotide sequence has been assigned GenBank Accession No. EU100973. Pathogenicity was confirmed through inoculations by gently pressing diseased leaves onto leaves of healthy C. officinalis plants. Five plants were inoculated. Five noninoculated plants served as control. Plants were maintained in a greenhouse at temperatures ranging from 20 to 26°C. Eleven days after inoculation, typical symptoms of powdery mildew developed on inoculated plants. Noninoculated plants did not show symptoms. The pathogenicity test was carried out twice. To our knowledge, this is the first report of powdery mildew on C. officinalis in Italy. C. officinalis was previously described as a host to Sphaerotheca fuliginea (synonym S. fusca) in Great Britain (4) as well as in Romania (3). Voucher specimens are available at the AGROINNOVA Collection, University of Torino. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (3) E. Eliade. Rev. Appl. Mycol. 39:710, 1960. (4) F. J. Moore. Rev. Appl. Mycol. 32:380, 1953.

First Report of Powdery Mildew Caused by an Oidium sp. on Spiraea japonica in Italy.

Spiraea japonica is an old-fashioned shrub widely grown in Italy in parks and gardens. In the summer of 2003, severe outbreaks of a previously unknown powdery mildew were observed in some parks and gardens in the city of Torino (northern Italy). Infected leaves became covered on both sides with a white mycelium. As the disease progressed, infected leaves turned reddish and eventually dropped prematurely. The presence of powdery mildew infections on leaves sometimes caused their distortion and growth reduction. Frequently, mycelium was observed also on the stem. Conidia were hyaline, ellipsoid, cylindrical, or dol iform, born in chains, measured 21.0 to 38.4 × 10.8 to 14.4 mm, and showed fibrosin bodies. Cleistothecia were not observed. The pathogen was identified as Oidium subgenus Fibroidium (1,2). Pathogenicity was confirmed by spraying leaves of healthy potted S. japonica plants with a conidial suspension (105 conidia per ml) prepared in sterile water from diseased leaves. Three plants were inoculated and three noninoculated plants served as control. The artificial inoculation was carried out twice. After artificial inoculation, plants were maintained in a growth chamber at 25°C. After 20 days, powdery mildew symptoms developed. Microsphaera alni and Podosphaera oxyacanthae were described as causal agents of powdery mildew on S. japonica in the United States (3), while Sphaerotheca spiraeae was considered the causal agent of a powdery mildew observed in Japan (4) and more recently in Poland. References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (2) R. T. A. Cook et al. Mycol. Res. 101:975, 1997. (3) P. Pirone. Diseases and Pests of Ornamental Plants. John Wiley and Sons, NY, 1978. (4) K. Sawada. Rev. Appl. Mycol. 31:577, 1952.

Engineering and design in the bioelectrochemistry of metalloproteins.

Engineered metalloproteins offer interesting systems for electrochemical studies of protein structure/function and their applications in nanobiotechnology. Scanning probe microscopy and cyclic voltammetry of engineered metalloproteins and electrodes have proved to be a powerful combination of tools contributing to the field of bioelectrochemistry. The ability to engineer tags, such as histidine tags and biotin-acceptor peptides, and to site-specifically introduce cysteine residues enabled the creation of ordered immobilised protein structures that can be characterised both electrochemically and topographically. Gene fusion and de novo combinatorial synthesis of metalloproteins are emerging to provide structures with the desired electrochemical properties.

[Treatment of intraepithelial cervical neoplasia with REP Radiosurgical Excision Procedure)]. / Intervento di REP (Radiosurgical Excision Procedure) nel trattamento della neoplasia cervicale intraepiteliale.

Radiosurgical cervical excision has been performed in 59 patients affected by CIN of various stages. This non-traumatic method uses 3.8 MHz radio waves to cut and/or coagulate without postoperative pain and tissue destruction. The radiosurgical excision has been performed in local anesthesia by a loop electrode or microneedle according to the seat of the lesion. The therapy was "effective" in 98% of cases (58/59) whereas the "incomplete excision" of CIN was observed in 6.7% (4/59). Complications (postoperative and late bleeding) occurred in 3.2% of the patients (2/59). Therefore, the Radiosurgical Excision Procedure (REP) can be considered an easy technique in ambulatory surgery; this method is effective in the treatment of CIN with either therapeutic and economic advantages and guarantee the functional and anatomic integrity of the cervix.

Spectroscopic properties of an engineered maltose binding protein.

The maltose binding protein (MBP) has been site specifically labelled with a nitrobenzoxadiazole (NBD) group following mutation of a serine to a cysteine residue at position 337. The resulting protein shows a large ligand (maltose or beta-cyclodextrin) dependent increase in its steady-state fluorescence intensity. Analysis of the static (intensity and anisotropy) and dynamic (lifetime distributions) fluorescence of the NBD label as well as the tryptophan residues in both ligand-bound and ligand-free states of this molecule reveals complex multi-component decays that are interpreted in terms of a ligand-induced solvent shielding mechanism. In the context of the known crystal structures of the various forms of the maltose-binding protein (MBP), ligand-dependent changes in both the fluorescence parameters as well as the circular dichroism spectra of the NBD group are interpreted by a twisted intramolecular charge-transfer (TICT) mechanism, wherein ligand binding locks the NBD group into a conformation that prevents efficient relaxation of the excited state.

[Endometrial carcinoma: an increasing neoplasm. Screening and early diagnosis: proposal for a protocol]. / Il carcinoma dell'endometrio: una neoplasia in aumento. Screening mirato e diagnosi precoce: proposta di protocollo.

Endometrial cancer is the most common cancer of the genital tract and it represents 10% of all cancers diagnosed in women. A protocol for screening and early diagnosis of this tumor has been designed by the authors. All asymptomatic women with risk factors undergo transvaginal sonography. The value of endometrial thickness suggests the need for endometrial sampling. On the contrary, such a procedure is always combined with transvaginal sonography in case of women with atypical genital bleeding. The aim of the protocol is to verify the efficacy of transvaginal sonography and endometrial sampling as combined procedures for screening and early diagnosis of endometrial cancer.

[Carcinoma of the endometrium: a neoplasm on the increase. Its staging and treatment: a proposed protocol]. / Il carcinoma dell'endometrio: una neoplasia in aumento. Stadiazione e trattamento: proposta di protocollo.

Endometrial cancer staging is obtained by the analysis of the surgical-pathological parameters. Therefore, surgery represents the first approach to this neoplasia. Surgical-pathological data analysis informs us of the main prognostic factors and the spread of the disease. On the basis of these findings, the risk of recurrence can be estimated and adjuvant treatments can be planned. A protocol for staging and treatment of endometrial carcinoma has been designed by the authors. After surgery, adjuvant therapy (chemotherapy or radiotherapy or the two modalities combined) is suggested by analysis of the prognostic factors. The aim of the protocol is to increase the relapse-free survival and to improve the quality of life of endometrial cancer patients.

Pure Leydig cell tumour (hilus cell) of the ovary: a rare cause of virilization after menopause.

A 58-year-old postmenopausal woman with high plasma testosterone levels and virilization, as demonstrated by hirsutism and alopecia, is presented. Urinary 17-ketosteroids and 17-hydroxycorticosteroids as well as the computed axial tomography scan of the adrenal glands were normal. Although no pelvic mass was detected by sonography or pelvic examination, the patient was found to have small pure Leydig cell tumour of the left ovary. Following total abdominal hysterectomy and bilateral salpingo-oophorectomy, the patient had regression of the hirsutism, and the plasma testosterone dropped to normal level.

Strongyloides stercoralis hyperinfection in a case of idiopathic pulmonary fibrosis.

The Strongyloides stercoralis infection is a prominent cause of death in many areas of the world. A 64-year-old man with pulmonary fibrosis was admitted to the hospital because of progressive shortness of breath and increasing cough. The patient had been previously admitted for his illness and had a remarkable immunosuppression due to the use of steroids (CD4+ lymphocytes = 200 x 10(6)/l). Repeated sputum and stool studies were diagnostic for strongyloidiasis. The patient died suddenly from severe cardiorespiratory failure while he was under mebendazole treatment (100 mg b.i.d.). Strongyloidiasis occurs mostly in immunocompromised hosts, as in patients with chronic pulmonary diseases on long-term treatment with corticosteroids, oncology patients under treatment and patients with AIDS.

[Giant pedunculated clustered uterine fibromyoma in hyaline and cystic degeneration. Limitations of clinical and instrumental diagnosis]. / Fibromioma uterino gigante peduncolato a grappolo in degenerazione ialina e cistica. Limiti della diagnostica clinica e strumentale.

The authors report a case of a 46-year-old woman who was checked by integrated ultrasound examination, i.e. transabdominal and transvaginal, and by abdominopelvic CAT. The examinations, ascertained the presence of a probably malignant, voluminous, ovarian, multilobate abdominopelvic neoplasia. On opening the belly, it was found that the tumefaction was a voluminous, pedunculate uterine myofibroma in clusters starting from the rear isthmian wall while the adnexa were normal. The histological examination confirmed that it was a "fibromyoma in hyaline and cystic degeneration". The authors conclude that although the image diagnostics has reached an extremely high level of precision, it is not yet possible to make an absolutely certain diagnosis, either on the organ from which a pelvic neoplasia originates or on its biological nature.

[The possibility of using sulprostone during laparoscopic myomectomy in uterine fibromyomatosis. Preliminary studies]. / Possibilità d'impiego del sulprostone in corso di laparomiomectomia nella fibromiomatosi uterina. Indagini preliminari.

The authors considered sulprostone effects, PGE2 synthetic derivative, on the hematic loss during laparomyomectomy. This drug is been used by intravenous way during the operation in 84 patients affected with uterine myomatosis. The effectiveness and tolerance of the drug is thus evident in the reduction of the hematic intraoperative loss.

[Ultrasonographic monitoring of the endometrium in post-menopausal women asymptomatic or undergoing hormone replacement therapy]. / Monitoraggio ecografico dell'endometrio in donne in post-menopausa asintomatiche od in terapia ormonale sostitutiva (HRT).

This study evaluated the reliability of ultrasonography in the post-menopausal period for the screening of endometrium pathology in women who are non-symptomatic or undergoing hormone replacement therapy. A total of 152 women were examined, 76 were non-symptomatic and 76 were undergoing estro-progestational therapy. As in previous studies on the subject, we took 0.8 cm as the thickness limit for the endometrium; over this limit closer investigations are considered necessary. In the non-symptomatic women, a strong correlation between the thickness of the endometrium and hormonal state was noted; it was not possible to differentiate the proliferative-secretory endometrium from the hyperplastic one. For women under HRT ultrasonography proved to be a good method for monitoring the endometrium as it allows for an adequate evaluation of its thickness throughout the different phases of sequential hormone treatment.

Engineering the maltose binding protein for reagentless fluorescence sensing.

This paper describes a mutant of the maltose binding protein (MBP) in which the serine residue at position 337 is replaced by a cysteine residue using site-directed mutagenesis. The mutant MBP has an approximately 2-fold lower affinity for maltose, and the cysteine residue can be modified with 4-[N-(2-(iodoacetoxy)ethyl)-N-methylamino]-7-nitrobenz-2-oxa-1,3-diazole (IANBD) and 6-acryloyl-2-(dimethylamino)-naphthalene (acrylodan). This combined genetic and chemical modification places the fluorophores close to the maltose binding site such that when the ligand is added the fluorescence intensity of the labels increases by 60-180% over that of the ligand-free form. This change is consistent with the fluorophores being buried when the conformation of the protein changes with maltose binding. Titration of the labeled mutant proteins yields dissociation constants for maltose of 62 +/- 0.2 and 0.8 +/- 0.01 microM respectively for the IANBD and acrylodan modifications. The application of this strategy of combined genetic and chemical modification to the development of reagentless fluorescence sensing is discussed.

[Severe anomaly of fetal limbs after early and failed voluntary interruption of pregnancy]. / Severa anomalia degli arti fetali dopo precoce e fallita interruzione volontaria di gravidanza.

The authors report the case of a patient who underwent a failed legal abortion at the 7th week of pregnancy via vacuum aspiration. A follow-up ultrasound survey done at the 14th week demonstrated the continuance of pregnancy and the presence of a serious fetal deformity consisting of the amelia of the two upper limbs, complete phocomelia of the right lower limb and distal phocomelia of the left lower limb. In the light of recent findings of a high incidence of fetal limb abnormalities in women subjected to chorionic villus sampling, before the 9th week of gestation, the Authors hypothesize that chorionic villus sampling, occurring during the abortion attempt, could probably be the cause of the fetal limb abnormalities in the present case study.

Conservative treatment of CIN: a review.

This review examines the indications for and the various therapeutic techniques used in the management of CIN. There is an extensive review of the literature. We believe that conization should be the long-term therapy of choice in over 90% of patients with CIN.