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Objectives The aim of this survey was to evaluate the different surgical approaches for women with high-grade vaginal intraepithelial neoplasia (HG-VaIN) used in 8 hospitals in central and northern Italy in the last 20 years. In particular, the baseline characteristics of the patients and factors potentially leading to excisional treatment rather than ablation were considered. Moreover, the clinical outcome of patients treated for HG-VaIN (disease persistence or recurrence and progression toward invasive vaginal cancer) was analyzed. Materials and methods The medical records of all women initially diagnosed with HG-VaIN and subsequently treated in 8 Italian hospitals from January 1996 to December 2016 were analyzed in a multicenter retrospective case series. Results Among the 226 women included, 116 (51.3%) underwent ablative procedures and 110 underwent excisional surgery (48.7%). An ablative procedure was preferred in cases where multiple lesions were found on colposcopic examinations. Physicians decided more frequently to perform excisional procedures in women with menopausal status, high-grade referral cervical cytology, previous hysterectomy for human papillomavirus-related disease, or VaIN 3 on colposcopic-guided biopsy. Conclusions The surgical treatment of HG-VaIN should be tailored according to the clinical characteristics of each woman and each lesion. However, in potentially high-risk cases (VaIN 3, previous hysterectomy for human papillomavirus-related disease, and menopausal women) or in those cases in which an occult invasive disease cannot be ruled out, an excisional approach should be preferred.In any case, long-term follow-up is advisable in women treated for HG-VaIN.

A retrospective study was conducted on patients subjected to laparoscopic myomectomy at our institution from January 2017 to December 2018 to identify predictive factors of blood loss. Two multiple regression models were run to predict intraoperative blood loss and haemoglobin drop. Predictors of an increased intraoperative blood loss and haemoglobin drop were the presence of three-four fibroids at ultrasound (+47 ml

OBJECTIVES The aim of the present study was to observe the trend of length of cone excisions in women treated with cervical excision procedure in five institutions of Central and Northern Italy. MATERIALS AND METHODS A multicenter retrospective cohort study was conducted on women who underwent a cervical excision procedure between January 2006 and December 2014. The pertinent clinical, histopathological, and sociodemographic characteristics of each woman were collected. In particular, the length of the cone specimen was evaluated and all of the factors that potentially influenced the length of excision were considered. RESULTS A total of 1482 women who underwent a cervical excision procedure from January 2006 to December 2014 were included. A mean (SD) cone length of 12.9 (5.0) mm was reported, and mostly, a significant decrease during the whole study period emerged. Age (r = 0.1, p = .007) and preoperative diagnosis of glandular lesions (r = 0.1, p < .001) were significantly related to the length of cone excision on multivariate analysis. Compared with the carbon dioxide laser excisional procedure, loop electrosurgical excision procedure showed a negative correlation with the length of excision (r = -0.2, p < .001). CONCLUSIONS During the study period, a significant decrease in the length of cone excision was observed, probably reflecting the gynecologists' acquired awareness of the increased risk of adverse obstetric outcomes for future pregnancies in the case of wide cone excisions.

ObjectiveTo evaluate the risk of progression to high-grade squamous intraepithelial lesion (HSIL) (CIN2-3) or invasive cancer in women with histopathological diagnosis of low-grade squamous intraepithelial lesion (LSIL) (CIN1), managed in a long-term observational approach up to 5 years.DesignRetrospective cohort study.SettingFour tertiary referral hospital.Participants434 women with adequate colposcopy and complete colposcopic charts were included in the present analysis. Women with glandular lesions on the referral cytology or previous diagnosis of cervical dysplasia or invasive cervical cancer or with synchronous vaginal, or with HIV infection or immunodepression were excluded.Primary and secondary outcome measuresThe main study outcome was the rate of progression to histopathological HSIL (CIN2-3) or invasive cancer at any time during 5 years of follow-up. The possible risk factors were also evaluated. As secondary outcome, we analysed the possible risk factors at the 24-month evaluation for histopathological HSIL (CIN2-3) or invasive cancer progression between 2 and 5 years from initial diagnosis.ResultsA progression to histopathological HSIL (CIN2-3) was found in a total of 32 (7.4%) cases during 5 years of follow-up. A histopathological diagnosis of HSIL (CIN3) was found in four patients (0.9%) and no case of invasive cancer was detected. High-grade cytology at inclusion and the presence of a positive high-risk human papillomavirus (HR-HPV) DNA test at 2 years from inclusion maintained a significant correlation with the risk of histopathological progression to HSIL (CIN2-3).ConclusionsThe results of our study showed a low rate (7.4%) of histopathological progression to HSIL (CIN2-3) in women with LSIL (CIN1) diagnosis during long-term follow-up up to 5 years. In case of positive HR-HPV DNA test at the 2 years evaluation an excisional treatment could be the preferred choice to prevent progression to HSIL (CIN2-3) in the following years, preferring a continuation of follow-up in case of HR-HPV DNA negative result.

OBJECTIVE The aim of the study was to evaluate the association between colposcopic features, age, menopausal status, and overtreatment in women subjected to "see-and-treat" approach, to identify subgroups of patients in which this approach could be more appropriate. MATERIALS AND METHODS Retrospective multicentric cohort study conducted on women older than 25 years, with a high-grade squamous cytological report and a visible squamocolumnar junction, in which colposcopy and the excisional procedure were performed at the same time without a previous cervical biopsy (see and treat). Overtreatment was defined as histopathological finding of cervical intraepithelial lesion grade 1 or normal tissue. RESULTS Among the 254 included patients, the overall overtreatment rate was 12.6%, whereas in women with a grade 2 colposcopy, it was 3.2% and, in women with grade 1 colposcopy, it was 22.0%. Among the considered factors (age, menopause, and grade 1 colposcopy), only a positive association with overtreatment and grade 1 colposcopy emerged (odds ratio = 8.70, 95% CI = 2.95-25.62, p < .001). CONCLUSIONS See and treat may be appropriate in women older than 25 years with a visible squamocolumnar junction and a high-grade squamous cervical cytology. Patients need to be informed about the higher risk of overtreatment in case of a grade 1 colposcopic impression, which however may still be considered acceptable. Patient's age and menopausal status should not influence the decision to propose a see-and-treat approach.

Convectional laser-wave-communication-systems canalize only the linear momentum in time (or mutually in space) domain, therefore neglecting all the non-linear signal power related to the angular momentum and losing the related information, whilst in time-space-domain the power-information is carried by the spin-angular-momentum (SAM) the orbital-angular-momentum (OAM) and the linear-momentum (LM) related to the signals. This paper concerns with an innovative technique which uses the non-linear-power of the beam-waves-modes to canalize/extract simultaneous independent signals, in Coulomb gauge conditions and paraxial accuracy, to enable information space-time-domain-canalization with extremely large bandwidth regardless of the given frequency carrier. The paper illustrates verification tests results developed using an EM or optical laser ground-communication-system implementing a health-care-tele-assistance and patient health-care-tele-examination in data, audio and video star-link configuration using any UHF/VHF-frequency-carrier.

The aim of this study was to assess the vitamin D serum level in women with ovarian endometriosis; specifically, a possible correlation between the dimensions of ovarian endometriomas and vitamin D serum levels was evaluated.This was an observational study of childbearing-age women diagnosed with singleton ovarian endometrioma from January 2015 to December 2015. Women diagnosed with multiple ovarian endometriomas or extraovarian endometriosis were excluded.Forty-nine women constituted the initial study cohort. In these women, the mean (±SD) 25-OH-D3 serum level was 22.0 (±8.9) ng/ml, and 42 of them (85.7%) were diagnosed with hypovitaminosis D. In the "hypovitaminosis D women", the mean (± SD) diameter of ovarian endometriomas was 40.2 ± 22.6 mm, while in the "normal vitamin D serum level women" it was 26.7 ± 12.1 mm (p = 0.1). However, a significant linear correlation between 25-OH-D3 serum level and the diameter of ovarian endometriomas was found (r = -0.3, p = 0.03).We found a relatively high rate of women with ovarian endometriosis and hypovitaminosis D. Interestingly, a significant linear correlation between 25-OH-D3 serum levels and the diameter of ovarian endometrioma emerged.

The effects of different growth conditions (ventilated and closed vessels, medium with 0, 15 and 30 g dm−3 sucrose) during proliferation of donor quince (Cydonia oblonga Mill.) shoots (stage I) on net photosynthetic rate and soluble sugars content were evaluated. In order to assess the influence of these physiological parameters on morphogenesis, leaf explants harvested from donor shoots were induced to form somatic embryos and adventitious roots under ventilated and closed Petri dishes (stage II). Natural ventilation and low sucrose contents (0–15 g dm−3) promoted the photosynthetic rate of quince shoots whereas biomass accumulation was the highest in those shoots cultured with 30 g dm−3 sucrose in both vessel types and 15 g dm−3 sucrose under natural ventilation. Increasing sucrose content in the medium induced greater accumulation of sucrose in leaf tissues of donor shoots. The content of reducing sugars was higher than that of sucrose, and it appeared to be higher in shoots cultured under natural ventilation compared to those in closed vessels. Somatic embryogenesis and root regeneration were influenced by stage I and II treatments. A significant correlation between sucrose content in the leaves of donor shoots and the number of somatic embryos regenerated was found, suggesting that identification of biochemical and physiological characteristics of donor shoots associated with increased regeneration ability might be helpful for improving morphogenesis in plant tissue culture.

The effect of Azospirillum brasilense Sp245 on the micropropagation of three fruit rootstocks: Mr.S 2/5 plum (Prunus cerasifera×P. spinosa), GF 677 hybrid (Prunus persica×P. amigdalus), and MM 106 apple (Northen Spy×M1) was assessed. Rooted shoots were treated with 3×107 of Sp245 cells during transplantation from in vitro cultures to the acclimatization phase. After 60 days, growth parameters were positively affected by Sp245 inoculum. In the case of Mr.S 2/5, an increase in rootstock stem length and node number by 37% and 42%, respectively, compared to the control was noted. In the case of GF 677, the bacterial inoculum increased stem length and node number by up to the 75% and 65%, respectively, compared to the control. The inoculum did not exert on MM 106 for both parameters suggesting that the effects of Sp245 could depend on a specific clone-microbe association. In all cases, however, a higher vigor, consistent with a wider leaf area, was present in the inoculated plantlets demonstrating tha...

The effect of in vitro cultivation of donor shoots on subsequent morphogenesis in leaf explants of quince (Cydonia oblonga Mill.) clone BA29 was investigated. Proliferating donor shoots were cultured in ventilated or closed vessels under different photosynthetic photon flux densities (PPFD; 200 and 100 µmol m−2 s−1) with 0, 15, 30 g dm−3 sucrose. Shoots grown in ventilated vessels, especially with sucrose at 15 or 30 g dm−3, were better developed with fully expanded leaves compared to those in standard closed vessels. Leaves collected from pre-treated donor shoots were used to assess regeneration capacity. Somatic embryo production was highest in leaves harvested from shoots cultured in closed vessels with 30 g dm−3 sucrose and in ventilated vessels with 15 and 30 g dm−3 sucrose and under high PPFD which was, in comparison with the control treatment (closed vessel, 30 g dm−3 sucrose and low PPFD), about 2 to 2.5 times higher. A similar response was observed for root regeneration.

This paper presents the results of the interaction of red light (R) with blue light (B), applied to shoots of M9 apple (Malus pumila paradisiaca Schmid) rootstock, on the regulation of stem growth, apical dominance and branching. These results are compared with the active form of phytochromes (PHYs) under monochromatic and dichromatic light treatments. The inhibition of internode elongation was determined by PHY photoequilibrium, with the additional effect of B, by means of a separate photoreceptor. The development of phytomers appeared to be primarily due to the active form of PHY, with a marginal effect from B. Shoot growth, which combines internode elongation and development of the phytomer, was highest under R and lowest under B and far red light (FR), showing the largely positive role of PHY photoequilibrium. Under FR, reduced stem elongation was due to the very small number of phytomers formed. Apical dominance was inhibited, while branching was increased under R, corresponding to the highest values of PHY photoequilibrium determined. Apical dominance was increased and branching was reduced by B, indicating a complex interaction between PHY and B receptor. In the shoot cluster system, photomorphogenic behavior was dependent on the time of exposure to the different light qualities. The information gained from the study will be helpful in improving the set up of in vitro growth light conditions, and in providing useful insights into research of the development of the woody plant canopy, an important factor in ecological plant communities.

The extraordinary success of imatinib in gastrointestinal stromal tumor (GIST) represents a model for molecularly targeted therapy of solid tumors. Research is currently going to identify the molecular basis of mechanisms of action and drug resistance. For the optimal management of the patients treated, a multidisciplinary approach, including medical oncologists, surgeons, pathologists, and radiologists is needed. In this article, we reviewed recent advances in the clinical management of GIST patients treated with imatinib, and in the knowledge of the molecular mechanisms that are basic to imatinib effects.

The interaction between shoot proliferation and regulative action of light quality was investigated in terms of cluster growth and development. In the commercial laboratory, the shoot multiplication procedure is normally conducted by growing clusters of shoots during several sub-cultures. Thus in this work the regulative action of light quality was evaluated by analysing the shoot cluster proliferation rate, the evolution of axillary bud differentiation, the axillary shoot development on both initial explant stem (order 1 axis) and three representative lateral branches (order 2 axes), growing from the lateral buds located at the bottom of the initial explant. Experiments showed that light qualities regulate bud differentiation and interact with apical dominance, leading to strong control of axillary bud outgrowth, and, finally, resulting in a differentiated architectural formation of clusters and type of branching. Two independent actions were observed, one dependent on blue light and the other dependent on red light, evoking the role of specific photoreceptor systems in plant response. Blue and UV-A light increased the number of buds differentiated from the apical meristem, without affecting apical dominance. An opposite trend was induced by red, yellow and green light. The involvement of phytochrome became clear under red light, while possible specific receptors and/or specific interaction among the known photoreceptors can be postulated under the other two light qualities. Results confirm that in apple, proliferation is the product of the interaction between two biological responses, lateral bud differentiation, which can be defined as potential proliferation, and new shoot development, which is the effective proliferation, and both responses are regulated by light quality.

CO 2 concentration was monitored during three 15-day subculturing cycles in vessels containing actively proliferating plum cultures of Prunus cerasifera , clone Mr.S. 2/5. The effects of two photosynthetic photon flux density regimes: 50 ± 5 μmol m −2 s −1 and 210 ± 5 μmol m −2 s −1 were compared. Three distinct phases in the CO 2 trend were distinguished during each culturing cycle of both light treatments. In the first, occurring at the beginning of the culture cycle, the amount of CO 2 emitted by the cultures during dark periods was greater than that assimilated during the light periods. In the second phase, the opposite trend was detected, while in the third, the range of CO 2 day–night fluctuations increased or remained stable according to the number of explants per vessel. The treatment with 210 ± 5 μmol m −2 s −1 did not modify the CO 2 phase trend but induced more pronounced fluctuations in day–night CO 2 concentration. Under this light treatment, cultures reached CO 2 compensation point for a period as long as 48% of the total number of light hours, while under 50 ± 5 μmol m −2 s −1 , it was only 8%. The different range in CO 2 day–night fluctuations monitored throughout a subculturing cycle, appeared to be mainly induced by changes in culture growth dynamics.

The effects of three cytokinins, kinetin 4.5 μM (Kin), 6-benzylaminopurine 4.5 μM (BA) and N-phenyl-N′1,2,3- thiadiazol-5-yl-urea 4.5 μM (TDZ), and the effects of different treatment duration on the regeneration of adventitious shoots from quince (Cydonia oblonga Mill.) leaves were studied. In a first experiment, leaves treated with Kin for 0, 8, 16 and 24 d were transferred to BA or TDZ-containing growth medium. In a second experiment TDZ applied for 0, 4, 8, 12, 16 and 24 d was followed by BA. All treatments included 0.5 μM α -naphthaleneacetic acid (NAA). In the sequence Kin-BA, the production of adventitious shoots decreased and reddish-coloured nodular structures (RNS) of meristematic appearance increased with increasing duration of Kin treatment, while somatic embryo formation was optimal at 8 d. In the Kin-TDZ sequence, shoot production was initially pronounced, but it declined with increasing duration of the Kin treatment, while the number of roots, somatic embryos and RNS increased. TDZ-BA treatments induced marked shoot production, which gradually increased with increasing duration of TDZ treatment. The presence of TDZ and a long treatment duration appeared to be very important factors in inducing caulogenesis. Kin appeared to be effective in shoot induction but not in shoot development; the results of this work demonstrate that RNS were adventitious shoots blocked at an early developmental stage on account of insufficient cytokinin activity. BA was less effective than TDZ in inducing shoot regeneration. Finally, both Kin and BA applied after 2,4-D treatment promoted somatic embryo induction.

SummaryThe purpose of this work was to study the behaviour of Platanus acerifolia Willd. to propagation by cuttings. Over three consecutive years, rooting responses were evaluated in cuttings collected on different dates, treated with indole-3-butyric acid (IBA) or naphthaleneacetic acid (NAA) at different concentrations as hydro-alcoholic solutions, powder preparations or potassium salts, and exposed to bottom-heat for different periods of time. A high natural rooting capacity was observed, particularly in cuttings collected in December and January. Auxin treatments at concentrations between 0.025 – 6 mg ml–1, applied to cuttings planted in bottom-heated benches, were unsuccessful or had negative effects on the rooting of cuttings, while they appeared to be positive if applied to cuttings planted in the field. Bottom-heat enhanced the rooting of cuttings and its effect was directly related to the duration of treatment. Cutting survival was negatively influenced by prolonged heat treatment, while pre-trea...

Experiments were performed to evaluate the capacity of quince (Cydonia oblonga Mill.) leaves to regenerate somatic embryos and shoots and/or roots simultaneously. Leaves, treated for 2 d in liquid medium containing 2.5 mg dm−3 2,4-dichlorophenoxyacetic acid were cultured for 0, 3, 6, 9, 12, 15, 18, 21 d on a gelled medium supplemented with 1 mg dm−3 kinetin (Kin) and 0.1 mg dm−3 naphthalenacetic acid (NAA) and were transferred to a medium either without growth regulator (GR-) or containing 0.6 mg dm−3 6-benzylaminopurine (BA) + 0.2 mg dm−3 gibberellic acid (GA3) + 0.06 mg dm−3 indole-3-butyric acid (IBA) (GR+). Leaves producing somatic embryos (SEs) only, or adventitious roots (Rs) only, or SEs+Rs simultaneously, were detected on GR- culture medium; on GR+ medium, leaves producing adventitious shoots (Ss) only, SEs+Ss or SEs+Rs+Ss simultaneously, also appeared. Leaves producing both Ss+Rs were never detected. Proportions among the various types of regenerating leaves varied according to both the length of Kin+NAA treatment and the presence or absence of GR in the transfer medium. The greatest variations, both on GR− and on GR+, took place within the first 9 d of culturing on Kin+NAA. After this period, no further substantial differences in the trend of each type of regenerating leaf were observed. The length of the treatment with Kin+NAA also modified the proportions between the different types of morphogenic structures.

The influence of the gaseous composition of the atmosphere inside culturing vessels on somatic embryogenesis and on adventitious root formation was investigated in the quince clone (Cydonia ablonga Mill.) BA29. Leaves taken from in vitro-grown shoots were cultured in glass Petri dishes and exposed to ventilation with atmospheric air (flow rate 25 ml min−1) for 0, 5, 10, 20, and 40 d. Twenty days of ventilation reduced the frequency of embryogenic leaves and a further decrease was observed after 40d of treatment. Conversely, adventitious root formation in the ventilated dishes was never different from the untreated cultures. In a second test, leaves were incubated in atmospheres containing different levels of oxygen (0, 5.0, 10.0, and 21.0%) or carbon dioxide (0, 0.04, 0.15, 1.5, and 3.0%). Anoxia conditions almost completely inhibited somatic embryo and adventitious root formation, but without compromising callus formation and explant viability. In contrast, embryo and root regeneration occurred even in totally CO2-free atmosphere. Oxygen seemed to influence somatic embryogenesis according to a quadratic response; a similar relationship was also observed for root regeneration. Instead, no clear trend could be inferred between embryo or root regeneration and CO2 levels. Furthermore, in dishes flushed with gas mixtures containing oxygen or carbon dioxide somatic embryo formation was almost always lower than in confined dishes. A different result was observed for root regeneration, since the number of roots was never lower than in the control and increased appreciably with 3.0% CO2. These results demonstrate that atmosphere composition of the culture head-space can influence somatic embryogenesis in quince. The finding that both vessel ventilation and atmosphere replacement with different gas mixtures reduced somatic embryo formation suggests that gaseous compounds, different from O2 an CO2, present in the gaseous environment may promote embryogenesis in this species.

The effects of NaCl and CaCl2 on shoot regeneration from quince (Cydonia oblonga BA L29 clone) leaves were investigated. Caulogenesis was induced on in vitro-grown leaves treated for 2d in liquid Murashige and Skoog (MS) medium with 11.3 μM 2,4-dichlorophenoxyacetic acid and cultured on MS gelled medium supplemented with 4.5 μM thidiazuron and 0.5 μM naphthaleneacetic acid. Three experiments were performed: in the first, we compared the effects of NaCl at 0, 25, 50, 100, and 200 mM in factorial combination with 3, 9, and 27 mM CaCl2. In the second, NaCl was tested at 0, 5, 10, 20, 40, and 80 mM with CaCl2 at 0.3, 1.0, and 3.0 mM. The third experiment was carried out with the same experimental design as the second one but replacing NaCl with Na2SO4. Shoot regeneration was evaluated after 50 d of culturing: 25 in darkness and 25 in white light. In the first experiment, shoot regeneration was very poor and was observed only at the lower salt concentrations. In the second experiment, the percentages of caulogenic leaves were much higher, but decreased with increasing NaCl concentration. The more pronounced negative effect of the highest NaCl concentrations appeared to be partly mitigated by CaCl2 at 1 and 3 mM. The presence of 3 mM CaCl2, in the experiment with Na2SO4, appeared to be even more effective in reducing the adverse effect of sodium stress on caulogenesis. This result was attributed to the lower Cl− concentration in the growth medium, which resulted from replacing NaCl with Na2SO4. NaCl applied at low concentrations (5 and 10 mM) in combination with 3 mM CaCl2 exerted a favorable effect on adventitious shoot regeneration. As regards the Na+ and Ca2+ interaction, when the Na+/Ca2+ ratio was below roughly 35 and 20, with NaCl and Na2SO4, respectively, at least 60% of leaves showed regenerating capacity, but optimal values of this ratio were not derived.

The effects of different macroelement combinations on somatic embryogenesis of quince (Cydonia oblonga Mill.) were tested. Leaves were excised from shoot cultures of quince clones and cultured on macroelement combinations of 8 different growth media. Callus production varied depending on the medium and the clone combinations. The influence of genotype and macronutrient combination on somatic embryo and root regeneration was also observed. Clone BA 29 showed the highest embryogenic properties and Murashige and Skoog-based medium appeared to be the most favourable for somatic embryo formation. Root regeneration was higher on Woody Plant Medium and Schenck and Hildebrandt-based media. Interactive effects between genotypes and macroelement combinations were also detected both for embryo and root formation. In all treatments, somatic embryos underwent early developmental arrest and failed to convert into plants. Differences in embryo and root regeneration observed among macroelement combinations may be ascribable to different levels of medium nitrogen and probably to the ratio between nitrate and ammonium.

The effects of 2,4-dichlorophenoxyacetic acid (2,4-D) concentration, length of induction period and light quality on leaf regeneration of quince clone BA 29 were investigated. After 2, 4 or 6 days of induction with 2.5 mg l−1 or 5.0 mg l−1 2,4-D, leaves were cultured under red, blue, red+blue, far-red+blue, white, far-red light or darkness conditions. Leaves thereby treated showed different responses, with respect to somatic embryogenesis, callus, red-nodular structures or roots. Callus production increased with increasing 2,4-D concentration and induction period, although it was not influenced by light quality; the only exception was far-red+blue light, which reduced callusing response. This result suggested involvement of the blue-absorbing photoreceptor system in the callus formation processes. A high regeneration of red-nodular structures with a meristematic appearance was also observed; from some histological characterizations, we presumed they were adventitious buds that were arrested at an early developmental stage. Red-nodular structures increased with decreasing 2,4-D concentration and induction period. In the regeneration of such structures, the blue-absorbing photoreceptor system appeared to have a negative effect but only at a low photoequilibrium value. In contrast, light quality which activated phytochrome induced an increment in regeneration, but the response did not vary for photoequilibrium values ranging from 0.43 to 0.86. For root regeneration, phytochrome seemed to be the only photoreceptor involved.

The effect of light quality on somatic embryogenesis in quince BA 29 was investigated. 2,4-D induced leaves were exposed for 25 days to the following light quality treatments: dark, far-red, far-red+blue, far-red+red, blue, white, red+blue, red. After a further 20 days of white light exposure, somatic embryo production was recorded. Somatic embryogenesis was highest in cultures subjected to red light treatment, and decreased progressively with the transition to red+blue and to white. Overall, embryogenic competence showed a correlation with photoequilibrium. Phytochrome appeared to be inductive although this effect was adversely influenced by the blue absorbing photoreceptor, in particular at low photoequilibrium. Independently of light treatments applied, somatic embryos frequently showed severe morphological abnormalities. Conversion of somatic embryos to plantlets was not observed.

SummaryThe influence of arbuscular mycorrhizal fungi on in vivo root initiation and development of Mr.S. 2/5 microcuttings was investigated. Micropropagated shoots of the Prunus cerasifera L. rootstock, clone Mr.S. 2/5, were placed in a sterile rooting mixture or inoculated with arbuscular mycorrhizal (AM) fungi. A part of the uninoculated microcuttings received auxin treatment (IBA 200 mg F1) or were supplied with a non-sterile sievate of mycorrhizal inoculum. Rooting percentage, root system morphology and growth increments of in vivo rooted microcuttings were evaluated. In vivo rooting of Mr.S. 2/5 shoots was obtained successfully but mycorrhizal inoculation did not show a positive influence on rooting ability of microcuttings. First harvest of plantlets was carried out very early, when mycorrhizal and non-mycorrhizal plants showed no significant growth differences. By this time, AM fungi had greatly increased the proportion of the root system present as higher-order laterals and branching intensity of ...

The effect of the photoperiod on some stomatal characteristics in various leaf lamina zones and in leaves of different age was studied on in vitro grown shoots of Prunus cerasifera clone Mr.S. 2/5, t Malus pumila Mill clone M9, and peach x almond hybrid clone GF 677. Stomatal density was highest in leaves exposed to continuous light and lowest with continuous dark. Photoperiod treatments supplying the same quantity of daily radiation but distributed over different cycles (4 h light and 2 h dark, 16 h light and 8 h dark) led to differing stomatal densities intermediate between those of the above light treatments. The light regime with the shortest light and dark periods was found to be most favorable to stomatal differentiation. Regardless of light treatment, stomatal density was found to be lower in the leaf lamina basal zone as compared to the median and apical zones, and decreasing from the 1st to 3rd to 5th leaf counting down from the apex. The photoperiod effect was also confirmed by a stomatal index. The stomatal axis ratio showed no interaction with the photoperiod but did highlight a tendency to rounder stomatal shape with increasing stomatal age.

We studied the effects of phosphate fertilization and inoculation with the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. and Gerd.) Gerdmann and Trappe, Glomus intraradices Schenck and Smith or Glomus viscosum Nicolson on shoot apical growth of plantlets that had been micropropagated from MM 106 apple (Malus pumila L.) and Mr.S. 2/5 plum (Prunus cerasifera Ehrh.) rootstocks. Unfertilized and non-mycorrhizal plantlets showed no apical growth during the post in vitro acclimation phase, whereas P fertilization induced early resumption of shoot apical growth. Growth enhancement and percentage of actively growing apices of mycorrhizal-inoculated plantlets were comparable to those obtained in plantlets fertilized with P. Furthermore, tissue P concentrations of mycorrhizal plantlets were similar to those of plantlets fertilized with P. We conclude that mycorrhizal inoculation can be used as a biotechnological tool to overcome blocked apical growth and to reduce chemical inputs, especially P inputs, to micropropagated fruit trees.

We compared root system morphogenesis of micropropogated transplants of Prunus cerasifera L. inoculated with either of the arbuscular mycorrhizal (AM) fungi Glomus mosseae or Glomus intraradices or with the ericoid mycorrhizal species Hymenoscyphus ericae. All plants were grown in sand culture, irrigated with a nutrient solution that included a soluble source of phosphorus, for 75 days after transplanting. Arbuscular mycorrhizal colonization increased both the survival and growth (by over 100%) of transplants compared with either uninoculated controls or transplants inoculated with H. ericae. Arbuscular mycorrhizal colonization increased root, stem and leaf weights, leaf area, root length and specific leaf area, and it decreased root length/leaf area ratio, root/shoot weight ratio and specific root length. Both uptake of phosphorus and its concentration in leaves were increased by AM infection, although the time course of the relationships between intensity of AM infection and P nutrition were complex and suggested a role for factors other than nutrition. The time course for the development of infection varied. It was most rapid with G. mosseae, but it was ultimately higher with G. intraradices. None of the treatments significantly affected the lengths of adventitious roots or the first-, second- or third-order laterals that developed from them. Arbuscular mycorrhizal colonization increased the intensity of branching in all root orders with the effect being most obvious on first-order lateral roots where the number of branches increased from under 100 to over 300 brances m(-1). As a result, although first-order laterals made up 55% of the root systems of control plants, the comparable value was 36% in AM-infected plants. In contrast, second-order laterals represented 25% of control root systems, but 50% of AM-colonized root systems. Glomus intraradices but not G. mosseae increased root diameter. Anatomical studies revealed no changes in the overall form of the root tip, although there were changes in the diameter of the root cap, cell numbers and cell size. Hymenoscyphus ericae increased the duration of the metaphase index. Both AM fungal treatments increased the concentrations of soluble proteins in root extracts and modified the protein profiles by the elimination and addition of protein bands detected by PAGE analysis. We conclude that AM fungal inoculation influenced processes in the root system at different levels, but not all effects were due to improved P nutrition or increased physiological age.

SummaryShoots of Mr. S. 2/5, a plum rootstock selected from Prunus cerasifera seedlings, were rooted in vitro at 100%, 90%, 80% and 70% relative humidity (r.h.). Decreases in r.h. led to increased resistance to water loss when the plantlets were subsequently exposed to a low environmental relative humidity. Statistically significant differences were observed after only 20 min of exposure. Differences increased progressively over time up to the end of the 90 min exposure period. In 80% and 70% r.h. treatments, the leaf waxy mesh was denser and presented thicker filaments. In addition, the reduction in r.h. induced a decrease in stomatal density; stomata were also more ellipsoidal and had a smaller opening. Net photosynthetic activity at the end of the in vitro rooting stage showed a slower rate in plantlets obtained from the 80% r.h. treatment compared with 100% r.h. plantlets; however, the total CO2 uptake over an 8 h period of illumination tended to be similar in plantlets from both r.h. treatments.

Biomass and length of intraradical and extraradical mycorrhizal mycelium under ambient (aCO2) and elevated (eCO2 ) atmospheric CO2 was investigated using a non-destructive in vivo experimental model system. Time-course experiments allowed measurements of intact extraradical mycelium spreading from mycorrhizal roots of Prunus cerasifera micropropagated plants inoculated with the arbuscular mycorrhizal fungus Glomus mosseae, in controlled environmental chambers. The length of extraradical mycelium was significantly increased at the highest CO2 concentration, ranging from 10.7 to 20.3 m at aCO2 and eCO2, respectively. The biochemical determination of mycelial glucosamine content allowed the evaluation of intraradical and extraradical fungal biomass, which were 2 and 3 times larger at eCO2 than at aCO2. Present data show that Glomus mosseae responds to increases of CO2 concentrations producing larger mycorrhizal networks which may potentially represent carbon sink agents in soil ecosystems.

The application of 4 h light followed by 2 h darkness (4/2 h), compared to 16 h light-8 h darkness (16/8 h). induced very different patterns of CO2 evolution from proliferating shoots of Prunus cerasifera in vitro. Under the former light/dark regime, fresh and dry weights of shoot clutures and the number of newly formed shoots were increased. Cultures under 4/2 h showed a higher photosynthetic capacity at an earlier stage of growth but this did not appear to be a factor in the enhanced growth. It is suggested that phytochrome may be involved in determining a different pattern of growth.

Des vitroplants deprunier (Prunus cerasifera Ehrh clone Mr.S.2/5) ont ete inocules post-vitro avec 4 champignons endomycorhizogenes: Glomus mosseae, G caledonium, G coronatum et Glomus souche A6, dans le but d'evaluer les parametres d'infection. Les champignons le plus infectieux (G mosseae) et le moins infectieux (G coronatum) ont ete respectivement selectionnes et leur capacite d'influencer la croissance de vitroplants de prunier a ete evaluee. Ces 2 champignons ont un effet positif sur la croissance des plants inocules []

Trials were conducted to study the effect of different photoperiods on in vitro growth of Mr.S.2/5 plum rootstock. Three photoperiods-16 h (control), 12 h and 8 h-were applied, with a PAR of 39 μmol m-2 sec-1. Tips collected from in vitro established shoots were used;ggrowth medium was MS with BA at 2.7 μM, GA3 at 0.7 μM and IBA at 0.3 μM.

Inoculation with Azospirillum brasilense Sp245 exerts beneficial effects on micropropagated plants of Prunus cerasifera L. clone Mr.S 2/5, as seen in the results of a comparative analysis of inoculated and non-inoculated explants, during both the rooting and acclimatation phases. The presence of Azospirillum brasilense Sp245 increased root system, root hair biomass production and apical activity. Although the presence of the bacteria had a positive effect on rooting, the addition of indolebutyric acid (IBA) to Murashige and Skoog (MS) medium was seen as indispensable in order to promote the rooting of explants. Aside from the promotion of plant growth, A. brasilense Sp245 protects plants against pathogen attacks, such as Rhizoctonia spp., with a plant survival rate of nearly 100% vs. 0% as seen in the negative control. The biocontrol effect of A. brasilense Sp245 on the fungal rhizospheric community has been confirmed by denaturing gradient gel electrophoresis (DGGE) profiles of the rhizospheric microbial community. This study indicates that A. brasilense Sp245 could be employed as a tool in plant biotechnology.

The purpose of this work was to acquire more information on the capacity of in vitro grown quince ( Cydonia oblonga Mill.) leaves to simultaneously regenerate somatic embryos, adventitious roots and shoots, and to evaluate the variations induced on regeneration response by treatments of different length with growth regulators. After 2 days of liquid treatment with 2,4-dichlorophenoxyacetic acid, the leaves were cultured for 0, 3, 6, 9, 12, 15, 18 and 21 days on gelled growth medium containing the basal components of Murashige and Skoog and kinetin (Kin) 4.5 μM + naphthaleneacetic acid (NAA) 0.5 μM. At the end of each treatment period, the leaves were cultured on a transfer medium in the absence or in the presence of a growth regulator combination represented by N 6 -benzylaminopurine (BA) 2.66 μM + gibberellic acid 0.58 μM + indole-3-butyric acid 0.3 μM. The culture period for all the treatments was fixed to 52 days. Simultaneous regeneration response of somatic embryos, adventitious roots and shoots differed according to the length of the kinetin + naphthaleneacetic acid treatment and to the absence or the presence of growth regulators in the transfer medium. In the absence of growth regulators, only somatic embryos and roots were regenerated; the number of somatic embryos increased from the 0 day to the 6–9th day of treatment with kinetin + naphthaleneacetic acid, while root regeneration showed the opposite trend. After the 9th day, the production of somatic embryos decreased up to the 15th day and then stabilized, while roots showed only very small reduction up to the end of the experiment. In the presence of growth regulators in the transfer medium, adventitious shoots were regenerated together with somatic embryos and roots. The highest number of shoots was observed when the leaves were directly transferred from 2,4-dichlorophenoxyacetic acid treatments to growth regulator medium. Increasing the length of kinetin + naphthaleneacetic acid treatment, shoot regeneration decreased and by the 12th day no more shoots were produced. Somatic embryo regeneration peaked on the 6th day of culture with kinetin + naphthaleneacetic acid and showed the same trend as in the absence of growth regulators in the transfer medium, while root formation tended to increase with prolonging the kinetin + naphthaleneacetic acid treatment.

Experiments were carried out to evaluate the effects of 4/2 light-dark cycles (4 h of light followed by 2 h of dark) on the rooting responses of shoots cultivated in vitro of the fruit tree rootstocks GF 677 (peach × almond hybrid), Mr.S. 2/5 (Prunus cerasifera), MM 106 (apple Nothern Spy × Paradise M1) and BA 29 (Cydonia oblonga). Under this light regime rooting percentage of GF 677, Mr.S. 2/5 and MM 106 shoots reached 100 % as in the control treatment (16/8), while in BA 29 shoots, short light-dark cycles increased rooting response by about 65 %. Moreover, the shoots of all rootstocks submitted to the 4/2 cycle showed an appreciable increase in root number and length, and an earlier root emergence of about 4 – 5 d compared to the 16/8 cycle. Finally, rooting percentage of BA 29 shoots submitted to the 4/2 light regime and treated with 0.2 mg dm−3 indolebutyric acid (IBA), was equal to that reported with 0.4 mg dm−3 IBA under the 16/8 regime, indicating that the former light regime also amplified the rhizogenic effect of auxin.

This paper presents HARE, a rule-based system addressing a section of the Italian law, and in particular the taxes to be paid to start legal proceedings. HARE uses an Italian version of Softlaw's STATUTE Expert. We will then shortly describe the main aspects of STATUTE Expert approach and some recent developments of this system.

Plum shoot proliferation was investigated in terms of two distinct processes: axillary bud differentiation and axillary shoot development. Results showed that light quality influenced bud differentiation and interacted with apical dominance in determining shoot outgrowth, resulting in a differentiated structure of shoot clusters and type of branching. Results suggested that blue light, acting through its photoreceptor, increased the number of axillary buds differentiated from apical meristem, but did not remove the apical dominance. Red light removed apical dominance, while reducing the formation of axillary buds; both events appeared to be dependent on the putative amount of phytochrome active form, and independent of light photon fluence rate. On the contrary, blue light action appeared to be dependent on photon fluence rate. In addition, apparent blue-red interactions related to photomorphogenic events fit an antagonistic model for branching regulated by light via cryptochrome and phytochrome photoreceptors. Our results show that the dynamics of shoot cluster development is the product of two events: the formation of new axillary buds and their release from apical dominance.