Treatment of multiple adjacent RT 1 gingival recessions with the modified coronally advanced tunnel (MCAT) technique and a collagen matrix or palatal connective tissue graft: 9-year results of a split-mouth randomized clinical trial.

OBJECTIVES: To evaluate t he long-term outcomes following treatment of RT 1 multiple adjacent gingival recessions (MAGR) using the modified coronally advanced tunnel (MCAT) with either a collagen matrix CM or a connective tissue graft (CTG). MATERIAL AND METHODS: Sixteen of the original 22 subjects included in a randomized, controlled split-mouth clinical trial were available for the 9-year follow-up (114 sites). Recessions were randomly treated by means of MCAT + CM (test) or MCAT + CTG (control). Complete root coverage (CRC), mean root coverage (MRC), gingival recession depth (GRD), probing pocket depth (PD), keratinized tissue width (KTW), and thickness (KGT) were compared with baseline values and with the 12-month results. RESULTS: After 9 years, CRC was observed in 2 patients, one in each group. At 9 years, MRC was 23.0 ± 44.5% in the test and 39.7 ± 35.1% in the control group (p = 0.179). The MRC reduction compared to 12 months was - 50.1 ± 47.0% and - 48.3 ± 37.7%, respectively. The upper jaw obtained 31.92 ± 43.0% of MRC for the test and 51.1 ± 27.8% for the control group (p = 0.111) compared to the lower jaw with 8.3 ± 46.9% and 20.7 ± 40.3%. KTW and KGT increased for both CM and CTG together from 2.0 ± 0.7 to 3.1 ± 1.0 mm (< 0.0001). There were no statistically significant changes in PD. CONCLUSION: The present results indicate that (a) treatment of MAGR using MCAT in conjunction with either CM or CTG is likely to show a relapse over a period of 9 years, and (b) the outcomes obtained in maxillary areas seem to be more stable compared to the mandibular ones. CLINICAL RELEVANCE: The mean root coverage at 12 months could not be fully maintained over 9 years. On a long-term basis, the results seem to be less stable in the mandible as compared to maxillary areas.

Fibroblast activation protein inhibitor (FAPI) PET for diagnostics and advanced targeted radiotherapy in head and neck cancers.

PURPOSE: Cancer-associated fibroblasts (CAFs) expressing fibroblast activation protein (FAP) have been associated with the aggressive nature of head and neck cancers (HNCs). These tumours grow diffusely, leading to extremely challenging differentiation between tumour and healthy tissue. This analysis aims to introduce a novel approach of tumour detection, contouring and targeted radiotherapy of HNCs using visualisation of CAFs: PET-CT with 68Ga-radiolabeled inhibitors of FAP (FAPI). METHODS: FAPI PET-CT was performed without complications prior to radiotherapy in addition to contrast enhanced CT (CE-CT) and MRI on 14 patients with HNC. First, for tissue biodistribution analysis, volumes of interest were defined to quantify SUVmean and SUVmax in tumour and healthy parenchyma. Secondly, using four thresholds of three-, five-, seven- and tenfold increase of FAPI enhancement in the tumour as compared with normal tissue, four different gross tumour volumes (FAPI-GTV) were created automatically. These were compared with GTVs created conventionally with CE-CT and MRI (CT-GTV). RESULTS: The biodistribution analysis revealed high FAPI avidity within tumorous lesions (e.g. primary tumours, SUVmax 14.62 ± 4.44; SUVmean 7.41 ± 2.39). In contrast, low background uptake was measured in healthy tissues of the head and neck region (e.g. salivary glands: SUVmax 1.76 ± 0.31; SUVmean 1.23 ± 0.28). Considering radiation planning, CT-GTV was of 27.3 ml, whereas contouring with FAPI resulted in significantly different GTVs of 67.7 ml (FAPI × 3, p = 0.0134), 22.1 ml (FAPI × 5, p = 0.0419), 7.6 ml (FAPI × 7, p = 0.0001) and 2.3 ml (FAPI × 10, p = 0.0001). Taking these significant disparities between the GTVs into consideration, we merged FAPI-GTVs with CT-GTVs. This resulted in median volumes, that were, as compared to CT-GTVs, significantly larger with FAPI × 3 (54.7 ml, + 200.5% relative increase, p = 0.0005) and FAPI × 5 (15.0 ml, + 54.9%, p = 0.0122). Furthermore, FAPI-GTVs were not covered by CE-CT-based planning target volumes (CT-PTVs) in several cases. CONCLUSION: We present first evidence of diagnostic and therapeutic potential of FAPI ligands in head and neck cancer. Larger studies with histopathological correlation are required to validate our findings.

Validation and discussion of clinical practicability of the 2022 graded prognostic assessment for NSCLC adenocarcinoma patients with brain metastases in a routine clinical cohort.

Introduction: The goal of this analysis is to validate the 2022 graded prognostic assessment (GPA) for patients with brain metastases from adenocarcinoma of the lung and to discuss its clinical practicability. Methods/material: 137 patients with adenocarcinoma of the lung were included in this analysis. The disease specific GPA for NSCLC, Lung-molGPA and the GPA for NSCLC adenocarcinoma were calculated. Overall survival was calculated for each GPA group. Additionally, expected and actual OS in the prognostic groups of the GPA available at the time of the patients' diagnosis was compared. Results: Median overall survival (OS) from diagnosis of brain metastases was 15 months (95% confidence interval (CI) 9.7-20.3 months). The median OS in the three individual prognostic groups was 7 months for GPA 0-1, 16 months for GPA 1.5-2, 33 months for GPA 2.5-3 and not reached for GPA 3.5-4 (p<0.001). Median survival times for the individual groups were similar to those published in the original GPA publication. Regarding the expected and actual OS when using the available GPA at the time of diagnosis there was an underestimation of survival of more than 3 months for all except the worst prognosis group. Conclusion: We were able to validate the 2022 GPA for NSCLC adenocarcinoma patients with brain metastases in a similar cohort from a non-academic center. However, the practical applicability regarding the expected median OS might be limited due to the constantly evolving treatment landscape and the consecutive improvement in overall survival.

Erythrocytes in sickle cell anemia are heterogeneous in their rheological and hemodynamic characteristics.

To understand the contribution to the pathophysiology of sickle cell anemia of the different erythrocyte density types present in the blood of these patients, we have studied the viscosimetric and hemodynamic characteristics of four major classes of hemoglobin SS erythrocytes. We have isolated reticulocytes, discocytes, dense discocytes, and irreversibly sickled cells (fractions I-IV) on Percoll-Renografin density gradients. Bulk viscosity was studied in a coneplate viscosimeter and the hemodynamic studies were performed on the isolated, artificially perfused mesoappendix vasculature of the rat (Baez preparation). Bulk viscosity measurements at shear rates of 230 S-1 demonstrate that when the cells are oxygenated, fraction I (reticulocyte rich) has a higher viscosity than expected from its low intracellular hemoglobin concentration. The rest of the fractions exhibit moderate increases in bulk viscosity pari-passu with the corresponding increases in density (mean corpuscular hemoglobin concentration). When deoxygenated, all cell fractions nearly doubled their bulk viscosity and the deoxy-oxy differences remained constant. The Baez preparation renders a different picture: oxygenated fractions behave as predicted by the viscosimetric data, but, when deoxygenated, cell fractions exhibit dramatically increased peripheral resistance and the deoxy-oxy difference are directly proportional to cell density, thus, the largest increases were observed for fractions III and IV. The differences between the rheological and the hemodynamic measurements are most probably due to the different sensitivity of the two methods to the extent of intracellular polymerization. These results also demonstrate that the hitherto unrecognized fraction III cells (very dense discocytes that change shape very little on deoxygenation) are as detrimental to the microcirculation as the irreversibly sickled cell-rich fraction IV. They may, however, induce obstruction by a different mechanism. As the extent to which these fractions are populated by erythrocytes varies considerably from patient to patient, the distribution function of cell densities in each sickle cell anemia patient might have consequences for the type of pathophysiological events occurring in their microcirculation.

The branched-chain amino acid transaminase 1 sustains growth of antiestrogen-resistant and ERα-negative breast cancer.

Antiestrogen-resistant and triple-negative breast tumors pose a serious clinical challenge because of limited treatment options. We assessed global gene expression changes in antiestrogen-sensitive compared with antiestrogen-resistant (two tamoxifen resistant and two fulvestrant resistant) MCF-7 breast cancer cell lines. The branched-chain amino acid transaminase 1 (BCAT1), which catalyzes the first step in the breakdown of branched-chain amino acids, was among the most upregulated transcripts in antiestrogen-resistant cells. Elevated BCAT1 expression was confirmed in relapsed tamoxifen-resistant breast tumor specimens. High intratumoral BCAT1 levels were associated with a reduced relapse-free survival in adjuvant tamoxifen-treated patients and overall survival in unselected patients. On a tissue microarray (n=1421), BCAT1 expression was detectable in 58% of unselected primary breast carcinomas and linked to a higher Ki-67 proliferation index, as well as histological grade. Interestingly, BCAT1 was predominantly expressed in estrogen receptor-α-negative/human epidermal growth factor receptor-2-positive (ERα-negative/HER-2-positive) and triple-negative breast cancers in independent patient cohorts. The inverse relationship between BCAT1 and ERα was corroborated in various breast cancer cell lines and pharmacological long-term depletion of ERα induced BCAT1 expression in vitro. Mechanistically, BCAT1 indirectly controlled expression of the cell cycle inhibitor p27Kip1 thereby affecting pRB. Correspondingly, phenotypic analyses using a lentiviral-mediated BCAT1 short hairpin RNA knockdown revealed that BCAT1 sustains proliferation in addition to migration and invasion and that its overexpression enhanced the capacity of antiestrogen-sensitive cells to grow in the presence of antiestrogens. Importantly, silencing of BCAT1 in an orthotopic triple-negative xenograft model resulted in a massive reduction of tumor volume in vivo, supporting our findings that BCAT1 is necessary for the growth of hormone-independent breast tumors.

Immunohistochemical investigation on the pattern of vimentin expression in regenerated and intact monkey and human periodontal ligament.

The expression of vimentin is well documented in the intact animal and human periodontal ligament (PDL), but there is limited information on the pattern of vimentin expression in the regenerated PDL. The aim of the present study was to investigate the pattern of vimentin expression in the regenerated and intact monkey and human PDL. A total of 12 chronic recession-type defects were created in three monkeys (Macaca fascicularis) and treated either with guided tissue regeneration (GTR), or with an enamel matrix protein derivative (EMD). After 5 months, the animals were sacrificed and specimens containing the defects and surrounding tissues were dissected free, decalcified in EDTA and embedded in paraffin. Sections were labelled immunohistochemically by using monoclonal antibody against vimentin (VIM 3B4). Twelve patients, each of whom displayed one deep intrabony defect scheduled for extraction were treated with GTR, EMD or combination of EMD+natural bone mineral (NBM). Following a healing period of 6 months, the teeth were extracted "en block" and immunohistochemically analysed according to the same protocol as described in monkeys. The results revealed that in both monkeys and humans the newly formed PDL was labelled similarly for vimentin to the intact (non-treated) PDL. In all specimens, the newly formed PDL was in continuation with the intact parts of PDL, thus suggesting that the mesenchymal cells capable of regenerating the attachment apparatus may have their origin in the intact PDL. In conclusion, the present findings indicate that (a) the reformed PDL displayed a similar expression of vimentin to the intact (original) PDL, and (b) the cells capable of regenerating new PDL and new cementum appear to be of mesenchymal origin and their source may be in the intact PDL.

Clinical and histologic evaluation of human intrabony defects treated with an enamel matrix protein derivative (Emdogain).

An enamel matrix protein derivative (Emdogain) has been recently shown to promote periodontal regeneration in experimentally created recession-type defects. However, only limited histologic data from human material are available concerning the healing of intrabony periodontal defects following treatment with Emdogain. The aim of the present study was therefore to present the clinical and histologic results following the application of Emdogain in intrabony defects. Two patients with marginal periodontitis and deep intrabony defects adjacent to teeth scheduled for extraction were treated with Emdogain. The postoperative healing phase was uneventful in both cases. At 6 months following treatment, newly formed cementum with inserting collagen fibers was found in both specimens. In one case, the new attachment formation was also accompanied by bone neoformation. The results of this human histologic study indicate that Emdogain possesses the potential to stimulate new connective tissue attachment formation in human intrabony defects.

GTR with bioresorbable membranes in the treatment of intrabony defects: a clinical and histologic study.

The aim of the present study was to evaluate clinically and histologically the treatment of intrabony periodontal defects with a bioresorbable membrane barrier. Fifty-two intrabony periodontal defects were treated according to the principles of guided tissue regeneration (GTR) with a bioresorbable membrane. Results were evaluated by assessing probing pocket depth, recession of the gingival margin, and clinical attachment level at baseline and at 1 and 2 years after therapy. Bone level changes were evaluated radiographically. The postoperative phase was uneventful in all cases. There was a mean probing pocket depth reduction from 8.4 to 3.6 mm, a mean increase of gingival margin recession from 1.5 to 3.0 mm, and a mean clinical attachment level change from 9.9 to 6.5 mm. Mean attachment gain was 3.4 mm. Two teeth scheduled for extraction were also treated with the same bioresorbable membrane. The histologic analysis 6 months after treatment revealed the formation of new connective tissue attachment and new alveolar bone in both cases. Based on the histologic findings it can be concluded that the clinical improvements following GTR with this type of bioresorbable membrane may represent, at least in part, true periodontal regeneration.

GTR with three different types of membranes in the treatment of intrabony periodontal defects: three-year results in sixty consecutive cases.

The aim of the present study was to evaluate the clinical results at 3 years following treating intrabony periodontal defects with different nonresorbable and bioabsorbable membrane barriers. Sixty intrabony periodontal defects were treated according to the principles of guided tissue regeneration (GTR). Twenty pockets were treated with Gore Resolut, a bioabsorbable membrane; 20 were treated with Gore-Tex, a titanium-reinforced membrane; and 20 with nonresorbable Gore-Tex membrane (all manufactured by Gore Regenerative Technologies, Flagstaff, AZ). The therapeutic results were evaluated by assessing probing pocket depth (PPD), recession of the gingival margin (GR), and clinical attachment level (CAL) at baseline, at 1 and at 3 years after therapy. The postoperative phase was uneventful in all cases. At 1 year after surgery, the results showed a mean PPD reduction from 9.42 mm to 3.35 mm (p < 0.0001) with Resolut; from 10.30 mm to 4.00 with titanium-reinforced Gore-Tex (p < 0.0001); and from 8.40 mm to 3.73 mm (p < 0.0001) with Gore-Tex membranes. The mean GR increased from 1.92 mm to 3.70 mm (p < 0.001) with Resolut; from 0.47 mm to 2.85 mm (p < 0.0001) with titanium-reinforced Gore-Tex; and from 0.73 mm to 2.15 mm (p < 0.0001) with Gore-Tex membranes. The mean CAL changed from 11.35 mm to 6.92 mm (p < 0.001) with Resolut; from 10.78 mm to 6.85 mm (p < 0.0001) with titanium-reinforced Gore-Tex; and from 9.13 mm to 5.87 mm (p < 0.0001) with Gore-Tex membranes. The clinical results at 3 years were not significantly different when compared with the 1-year results (p > 0.05). No significant differences existed between the mean changes in PPD, GR, and CAL in the three different test groups. Furthermore, one tooth scheduled for extraction for periodontal and prosthodontic reasons was treated with Resolut. Histological analysis 6 months after treatment demonstrated the neoformation of a connective tissue attachment and of new alveolar bone. This additional evidence thus proved that treatment with bioabsorbable membranes according to GTR principles delivers not only clinical improvement, but also histological periodontal regeneration.

[The effect of certain stages of periodontal treatment on the regeneration of periodontal tissues]. / A komplex parodontális kezelés egyes lépéseinek hatása a parodontális szövetek regenerációjára.

The regenerative potential of the periodontal tissues is relatively limited. The attachment loss has long been considered as an irreversible damage of the periodontium. Most of the conventional methods of the comprehensive periodontal treatment provided no convincing evidence of true new periodontal attachment formation. Most of the surgical and nonsurgical approaches achieved either secondary gingival recession and/or long epithelial attachement. The recently introduced guided tissue regeneration techniques can make the regeneration of the fibrous periodontal attachment and convincing clinical and histological evidences of new cementum and bone formation possible, as well as the regeneration of the perpendicular Sharpey's fibers fully embedded into the matrix of the appositionally formed new cementum and bone. The theories and clinical implications of these techniques are discussed and illustrated with clinical cases.

[Possibilities of reducing the sensitivity of the neck of the tooth]. / A fognyaki érzékenység csökkentésének lehetöségei.

Authors overview the etiology and pathomechanism of the root hypersensitivity. The most commonly used therapeutical aids and devices are discussed. They discuss the theory of action of the drugs used for controlling root hypersensitivity. In details the dental literature of devices used on the individual bases--included different fluoride compounds, stronciumchloride, and potassium nitrate--are reviewed. In the end the authors's own clinical experience and findings with the Sensodyne toothpaste (kindly donated by the Block Drug Co.) are presented.

[Importance of periodic follow up of periodontal diseases for the maintenance of periodontal health and the longevity of dental restoration]. / A fogágybeteg rendszeres ellenörzésének jelentösége a paradontium épségének megörzésében és a fogpótlás élettartamának meghosszabbításában.

The major objective of the author's investigation was to determine if the progression of attachment loss, the recurrence of active phase of periodontitis and development of new carious lesions can be controlled and prevented in individuals with prior history of aggressive periodontitis provided full mouth reconstruction and maintained at a proper level of oral hygiene. The results of a group on regular periodontal recall program were compared with the five years dental and periodontal findings on a matched group of patients having not been on regular maintenance program. The well maintained group of patients' annual recall data did not vary markedly. In five years basically the same amount of attachment apparatus was maintained. The reexamination of the non recall group at the end of the fifth year shoved a total deterioration of dentition due to the progression of periodontal disease and attachment loss. Especially those of rapidly progressing periodontitis cases shoved 70 to 100% attachment loss. This comparative study attempted to outline the insufficiency of our former dental philosophy of patient management and follow up. Authors emphasize the importance of the follow up, and maintenance of a very high level.

[A 3-year experience with guided tissue regeneration procedures]. / Az irányított szövetregenerációs technikákkal nyert három éves klinikai tapasztalatok.

The Guided Tissue Regeneration (GTR) procedures are promoting a clinically and radiologically as well as histologically verifiably periodontal attachment gain. The objective of the study was to evaluate the clinical efficacy of these GTR techniques. In the past four years different barrier membranes (Gore-tex, Resolut and Guidor) were used around 318 teeth of 196 patients. 169 periodontal defects of 140 patients were followed up at least for two years. 54 patient had chronic adult type periodontitis, 67 suffered with rapidly progressing periodontitis and 15 had different severe mucogingival lesions. 111 vertical bony defects, 43 Class II-III furcation lesions and 15 mucogingival lesions were surgically corrected. The average preoperative probing depth (PD) and the clinical attachment loss (CAL) of the vertical bony defects were 5.3 +/- 1.7 mm and 6.2 +/- 1.9 mm respectively. The PD of the deepest Class III furcation lesion was 11 mm. The average gingival recession of the mucogingival lesions was 4.5 +/- 1.1 mm. The GTR technique provided the best results in the Class II-III furcation lesions, where an average 2.4 +/- 0.9 clinical attachment gain was observed one year postoperatively. The GTR techniques provided an average 1.8 +/- 1.2 mm attachment gain in the vertical bony crater cases. In both groups of cases a marked gingival recession followed the healing and the periodontal regeneration. In this way the average reduction in the probing depth exceeded the average attachment gain by more that 1.5 mm. 1 year after the operation the average radiologic bone fill was about 0.9-1.2 mm. The resorbable barrier membranes resulted in clinically significant root coverage and an average 3.5 +/- 1.7 mm gain in the width of keratinized gingiva. The success or failure of our cases were mainly determined by the patient's compliance, the level of the postoperative professional and individual oral hygiene and the number of periodontal recalls. These findings are also underlining the importance of the high standard of oral hygiene in the postoperative periodontal regeneration.

Caudex growth and phenology of Cyathea atrovirens (Langsd. & Fisch.) Domin (Cyatheaceae) in secondary forest, southern Brazil.

The leaf production and senescence, formation and release of spores of Cyathea atrovirens (Langsd. & Fisch.) Domin were analysed based on the monthly monitoring of 50 plants growing in a secondary forest, in the municipality of Novo Hamburgo, in the state of Rio Grande do Sul, during the year 2004. The caudex height and number of mature and fertile leaves were recorded annually in 2004-09. In 2004, monthly production and senescence of leaves were concomitant, without total leaf abscission. Population synchrony at emergence (Z = 0.86) and leaf senescence (Z = 0.82) increased in spring but did not correlate with temperature and photoperiod. All individuals were fertile and the sporangia production and spore liberation presented higher and equal synchrony (Z = 0.84) respectively in spring and summer. Sporangia production was related with temperature and photoperiod, however taller plants did not produce more fertile leaves. Phenological events analysed were not influenced by precipitation, as expected for forests in non-seasonal climate. Over five years (2004-09), the annual mean caudex growth varied between 1.19 and 2.50 cm.year-1 and the plants appeared to have an ability to maintain a relatively stable amount of leaves throughout this period.

Effects of nickel on the fern Regnellidium diphyllum Lindm. (Marsileaceae).

The heterosporous fern Regnellidium diphyllum occurs in southern Brazil and some adjoining localities in Uruguay and Argentina. Currently it is on the list of threatened species in the state of Rio Grande do Sul. Anthropic alterations such as the conversion of wetlands into agricultural areas or water and soil contamination by pollutants may compromise the establishment and survival of this species. Nickel (Ni) is an essential nutrient for plants but increasing levels of this metal due to pollution can cause deleterious effects especially in aquatic macrophytes. Megaspore germination tests were performed using Meyer's solution, at concentrations of 0 (control), 0.05, 0.5, 1, 5, 10, 20, 30, 50 and 100 mg L(-1) of Ni. The initial development of apomictic sporophytes was studied using solutions containing 0 (control) to 4.8 mg L(-1) of Ni. A significant negative relation was observed between the different Ni concentrations and the megaspore germination/sporophyte formation rates. Primary roots, primary leaves and secondary leaves were significantly shorter at 3.2 and 4.8 mg L(-1) of Ni, when compared with the treatment without this metal. At 4.8 mg L(-1), leaves also presented chlorosis and necrosis. The introduction of pollutants with Ni in the natural habitat of Regnellidium diphyllum may inhibit the establishment of plants in the initial stage of development, a problem to be considered in relation to the conservation of this species.

Volumetric difference evaluation of registered three-dimensional pre-operative and post-operative CT dental data.

OBJECTIVE: The purpose of this study is to propose a complete methodology for automatically registering three-dimensional (3D) pre-operative and post-operative CT scan dental volumes as well as to provide a toolset for quantifying and evaluating their volumetric differences. METHODS: The proposed methodology was applied to cone beam CT (CBCT) data from 20 patients in order to assess the volume of augmented bone in the alveolar region. In each case, the pre-operative and post-operative data were registered using a 3D affine-based scheme. The performance of the 3D registration algorithm was evaluated by measuring the average distance between the edges of the registered sets. The differences between the registered sets were assessed through 3D subtraction radiography. The volume of the differences was finally evaluated by defining regions of interest in each slice of the subtracted 3D data and by combining all respective slices to model the desired volume of interest. The effectiveness of the algorithm was verified by applying it to several reference standard-shaped objects with known volumes. RESULTS: Satisfactory alignment was achieved as a low average offset of 1.483 ± 1.558 mm was recorded between the edges of the registered sets. Moreover, the estimated volumes closely matched the volumes of the reference objects used for verification, as the recorded volume differences were less than 0.4 mm(3) in all cases. CONCLUSION: The proposed method allows for automatic registration of 3D CBCT data sets and the volumetric assessment of their differences in particular areas of interest. The proposed approach provides accurate volumetric measurements in three dimensions, requiring minimal user interaction.

Biodiversity and spatial distribution of epiphytic ferns on Alsophila setosa Kaulf. (Cyatheaceae) caudices in Rio Grande do Sul, Brazil.

The extractive exploitation of the tree fern Alsophila setosa Kaulf. alters forest formations and diminishes the availability of micro-habitat for epiphytes. A survey of epiphytic fern communities on A. setosa at 16 study sites in different forest formations in the State of Rio Grande do Sul, Brazil, documented the occurrence of 31 species representing 16 genera and six families. The greatest richness of species occurred in Polypodiaceae (39%) and in the genus Asplenium L. (22%). Habitual holoepiphyte was the predominant ecological category, representing 61% of the species. Similarity analysis demonstrated heterogeneity in the composition of the epiphytic ferns at the study sites and indicated that the vegetation type is not the main factor for floristic difference. The lowest total specific richness (5) was recorded for the seasonal deciduous forest site. The majority of the sites presented similar averages of phorophyte height and epiphyte richness per caudex. In 25% of the sites the height of the host plants presented significant correlation with specific richness. Considering that the majority of the epiphytes are habitual and that some of them occur exclusively or preferentially on tree ferns, the maintenance of these host plants in the vegetation is essential for the conservation of epiphytic species.

Germination and sporophytic development of Regnellidium diphyllum Lindman (Marsileaceae) in the presence of hexavalent chromium.

Regnellidium diphyllum Lindman is a heterosporous fern, growing in aquatic environments and surrounding wetlands, which is assumed to be threatened by increasing water pollution and disappearance of its natural habitats. Among contaminants, hexavalent chromium - Cr(VI) - is known to be present in effluents from some leather tanning factories. Megaspore germination tests were performed using Meyer's solution, at concentrations 0 (control), 0.1, 0.5, 1, 5, 10, 15, 20, 30, 50, and 80 mg.L⁻¹, from a standard solution of Titrisol® 1000 mg.L⁻¹. The primary development of apomictic sporophytes was studied using solutions containing 0.025 to 4.8 mg.L⁻¹ of Cr(VI). The experiments were conducted in a growth chamber at 24 ± 1 ºC and for a 12-hour photoperiod under fluorescent lights, providing a nominal irradiance of 77 µmol.m⁻²/s. Significant differences in megaspore germination, with subsequent sporophytic development, were verified from 0.5 mg.L⁻¹ Cr(VI) concentration onwards. Growth of primary root and primary and secondary leaves was significantly reduced at 3.2 mg.L⁻¹ Cr(VI) concentration or higher. Considering the pollution from Cr(VI) in some areas of R. diphyllum natural occurrence, these data indicate that low reproductive rates and disappearance of populations are likely to occur in these situations.

Effects of 2,4-D on the germination of megaspores and initial development of Regnellidium diphyllum Lindman (Monilophyta, Marsileaceae).

Regnellidium diphyllum is considered as endangered, occurring in the State of Rio Grande do Sul, Brazil, and a few adjoining localities in Uruguay, Argentina and the State of Santa Catarina. It grows in wetlands frequently altered for agricultural activities. Herbicides based on 2,4-dichlorophenoxyacetic acid (2,4-D) are widely used in these fields. The effects of 2,4-D on the germination of megaspores and initial sporophytic development of R. diphyllum were investigated. Six concentrations of 2,4-D (0.32; 0.64; 1.92; 4.80; 9.60 and 19.20 mg.L-1), and the control (0.00 mg.L-1), were tested in vitro, using Meyer's medium. Cultures were maintained in a growth chamber at 24 +/- 1 degrees C, under artificial light with nominal irradiance of 110 micromol.m-2/s and 16 hours photoperiod. Megaspore germination was lower at 9.60 and 19.20 mg.L-1 of 2,4-D (56 and 48%, respectively), compared with the control (68%). Herbicide concentrations of up to 1.92 mg.L-1 did not significantly decrease the number of sporophytes formed. At 19.20 mg.L-1, no sporophytes were formed. The lengths of the primary root, primary and secondary leaves were greater at concentrations of 0.32 and 0.64 mg.L-1 of 2,4-D. Low concentrations of 2,4-D do not affect germination rates and initial development of R. diphyllum in a significant way. However, higher concentrations (9.60 and 19.20 mg.L-1) affect substantially the germination of the megaspores and interfere with the establishment of the species.