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Background/purpose: This randomized clinical trial was aimed at comparing two different means of delivering chlorhexidine digluconate (CHX) for plaque control during the 2 weeks following implant surgery., Materials and Methods: Twenty patients selected for implant therapy were randomly divided into two groups: 10 subjects used 15 ml of 0.12% CHX mouthrinse (control group) and 10 used 0.2% CHX spray (test group). Professional oral hygiene was carried out immediately before surgery. During the 14 days following surgery mechanical oral hygiene was performed only at the teeth not surgically involved. Plaque index (PI), stain index (SI), modified gingival index and taste alteration were assessed on the 7th and 14th day after surgery. The clinical parameters were evaluated at four tooth surfaces by a single examiner. Teeth proximal to surgical site and teeth not involved were statistically compared., Results: In both groups, the PI increased similarly, with respect to the baseline, at days 7 and 14. There was no significant difference between the two groups at either time point. On the contrary, in the control group, the SI increased significantly when compared with baseline over the 14 days both at teeth nearest to surgical sites and at not-involved sites. In the test group pigmentation was consistent only at teeth proximal to the surgical site. When considering not-involved sites, tooth staining was significantly lower in the test with respect to the control group., Conclusions: The present study indicates that the efficacy of CHX spray in the post-surgical control of dental plaque is similar to that of CHX mouthwash. Tooth staining, however, is significantly lower in the spray group at sites not surgically involved. These effects might be related to the route of CHX delivery, as well as the total dose administered that was significantly lower in the spray group with respect to the rinse group., (Copyright Blackwell Munksgaard, 2004)

Aim: To evaluate the apical seal of canals filled with a core-carrier obturator following root-end resection with and without a root-end filling., Methodology: Thirty single-rooted human teeth with single canals were used. Root canal treatment was performed and canals filled with a core-carrier obturation technique. The teeth were then randomly assigned to three groups (n = 10). In the first group root canal filling only was performed. After orthograde filling, the teeth of the second group were resected apically, perpendicular to the major axis of the root. In the third group after apical resection, a root-end cavity was prepared using ultrasonic diamond retrotips and the cavities filled with Super-EBA cement. During a period of 3 h and with a headspace pressure of 0.12 atm, methylene-blue solution was forced through a tube that was connected to the apical end of each tooth specimen. The coronal end of the tooth was connected to a capillary tube containing an air bubble. Leakage was evaluated by observing the distal displacement of the air bubble. The roots were then sectioned along their long axis. Using a stereomicroscope, linear dye infiltration at the dentine-cement interface was determined. Kruskal-Wallis and Mann-Whitney tests were used to compare the three groups., Results: Linear dye infiltration was significantly greater in root canals filled with the core-carrier obturators and resected apically (0.9 +/- 0.9 mm) when compared with those that had root-end fillings (0.2 +/- 0.4 mm). Air bubble displacement was not observed., Conclusion: Root-end filling improves the sealing of roots with core-carrier obturation following root-end resection.

The coronally advanced flap combined with a free connective tissue graft is a predictable method for achieving root coverage in buccal gingival recession. Nevertheless, this procedure conventionally requires involvement of a second surgical site; the latter is avoided by the proposed technique. Sixteen isolated gingival recessions (2.5 to 4.0 mm deep) were surgically treated with a coronally advanced flap associated with a connective tissue graft harvested from one adjacent papilla whose dimensions matched those of the exposed root area. Procedures were performed with the aid of a surgical microscope. Recession depth, probing depth, periodontal attachment level, and keratinized tissue width were recorded at baseline and 12 months after surgery. Mean recession moved from 3.38 +/- 0.72 mm at baseline to 0.13 +/- 0.29 mm at 12 months, a gain of 97.03%. In 13 of the 16 cases 12 months after surgery, the gingival margin was located at the CEJ or coronal to it, while in two cases the residual recession was less than 1.0 mm and in another case it was 1.0 mm. Mean periodontal attachment level was 4.72 +/- 1.00 mm at baseline and 1.03 +/- 0.59 mm at follow-up. Mean keratinized tissue increased from 1.25 +/- 0.75 mm to 3.47 +/- 0.87 mm. All differences between 12 months and baseline were statistically significant. No pockets were present at baseline, and this situation remained stable during the observation period. All 16 isolated recessions treated showed an excellent gain in root coverage without requiring a second surgical site and thus reducing patient morbidity.

Objective: This paper reports the results of a prospective multicenter clinical study on immediately fully occlusally loaded full-arch screw-retained prostheses with distal extensions (hybrid prostheses) supported by Osseotite implants inserted in edentulous lower jaws., Method and Material: Sixty-two patients were enrolled in four clinical centers. Three hundred and twenty-five Osseotite implants were inserted and occlusally loaded according to an immediate loading protocol. The temporary prosthesis was delivered 4 h from surgery. The final prosthesis was delivered after 6 months. Marginal bone loss was monitored from periapical radiographs using a computerized technique., Results: Two implants failed to integrate within 2 months of occlusal loading. A cumulative implant success rate of 99.4% was achieved for a period of 12-60 months postplacement (mean 28.6+/-14.1 (SD) months). Crestal bone loss around the immediately loaded implants was similar to that reported for standard delayed loading protocols., Conclusion: The results of this study suggest that the rehabilitation of the edentulous mandible by an immediate occlusally loaded hybrid prosthesis supported by five to six Osseotite implants represents a viable alternative treatment to classic delayed loading protocols.

Immediate loading of endosseous implants is becoming a widespread therapeutic procedure for the rehabilitation of patients with edentulous jaws. The purpose of this prospective clinical trial was to evaluate the long-term success rate of endosseous implants placed in the edentulous lower jaw and loaded on either the same day of surgery or the next day. Nineteen patients were enrolled in the study. Eleven patients, accounting for 64 implants, received their provisional prosthesis the same day of implant placement, and 8 patients, accounting for 52 implants, were rehabilitated the day after surgery. All patients were rehabilitated by a hybrid prosthesis supported by 5 to 6 Osseotite implants. Two implants failed in the group of patients who had their implants loaded the same day (96.9% success rate), whereas 1 implant failed in the other group (98.1% success rate). The overall implant success rate was 97.4%. All failures occurred within 2 months of function. No other complication was reported. The mean follow-up for this interim report was 37.8 +/- 16.5 months (range 8-65 months). Crestal bone loss was similar to that reported for standard delayed loading protocols. The results of this study suggest that the rehabilitation of the edentulous lower jaw by an immediate occlusally loaded implant-supported hybrid prosthesis is equally successful when loading is applied the same day or the day after implant placement. Immediate loading with 5 to 6 implant-supported prostheses represents a viable alternative treatment to classic delayed loading protocols.

Background: A growing flow of recent evidence indicates enamel matrix derivative (EMD, Emdogain) as a useful tool for the regeneration of periodontal tissues. This prospective clinical study aimed to evaluate the efficacy of EMD combined with surgical treatment of periodontal intra-bony defects, as compared with surgery alone, up to 24 months of follow-up., Methods: Twenty-four intra-bony defects were treated in 24 patients in a single clinical centre. Each defect had intra-bony depth (IBD) > or = 4 mm and probing pocket depth (PPD) > or = 6 mm. Patients were randomly assigned to either test or control group. Plaque index (PI), gingival index (GI), PPD and periodontal attachment level (PAL) were assessed at baseline pre-surgical examination at the site to be treated. Full mouth plaque score (FMPS) and full mouth bleeding score (FMBS) were also evaluated. Twelve patients were treated by simplified papilla preservation flap technique (control group), while 12 patients were treated with the same surgical technique plus EMD after ethylenediamine tetraacetic acid root conditioning (test group). Any probing at the involved sites was avoided in the first year post-surgery. Radiographs were taken at baseline, 12 and 24 months after surgery using customized bite blocks. Intra-bony defect depth (IBD) and angle (IBA) were measured from X-rays by a computer-aided technique. At 12 and 24 months post-surgery, FMPS, FMBS, PI, GI, PPD, PAL and radiographic IBD and IBA were assessed. The difference between each follow-up and baseline, and between groups at each follow-up was evaluated for the above parameters by standard statistical methods., Results: In both groups, clinical and radiographic parameters were improved at either 12 or 24 months when compared with baseline. The test group displayed better outcomes when compared with the control group for IBD, PPD, and PAL gain at 12 months, and only for PAL and IBD gain at 24 months. No adverse event related to the use of EMD was reported., Conclusions: The surgical procedure used in the present study, aiming for maximum preservation of the regenerative potential of periodontal tissues, showed per se excellent results. The use of EMD as an adjunct to periodontal surgery in the treatment of angular defects possibly enhances periodontal regeneration rate.

Immediate loading of dental implants can reduce treatment time and management concerns, thus increasing patient acceptance. This paper reports the authors' preliminary experience with partially edentulous patients having received non-occlusally loaded provisional restorations within 24 hours of surgery, compared to patients treated according to an early loading protocol. The present data suggest that implant supported fixed partial prostheses can be immediately non-occlusally loaded and restored with a predictability similar to early loaded implants.

The aim of this work is to develop an accurate finite element model able to reproduce a standard experimental set-up for the evaluation of mechanical failure of a dental implant system. The considered system is composed of a fixture, an abutment and a connecting screw. We analysed the behaviour of the implant system considering three different designs of the fixture, in order to establish which one provides the better mechanical behaviour. After the definition of the numerical models, loading conditions were selected in order to reproduce the same stress state found in previous mechanical failure tests. Preloading and functional loading conditions were simulated. The analysis of the numerical results shows that the structure yielding is due to the fixture neck plastic deformation, that increases the load eccentricity and then the bending stress on the connecting screw. Only slight differences were found between the three implant systems in the amount and distribution of stress. The model reproduces properly the implant systems and the experimental set-up. The goodness of the model can be summarised as: realistic geometrical structure, elastoplastic model for the material description, correct definition of the contacts and the existing tolerance among the different system components, reproduction of the preloading stress condition. The present study permitted to define a valid procedure for the realization of numerical models of implant systems.

Purpose: This article reports the preliminary data from a clinical study of immediately loaded, full-arch, screw-retained prosthesis with distal extensions (hybrid prosthesis) supported by Osseotite implants placed in the edentulous mandible., Materials and Methods: Fifteen patients who received 103 implants were enrolled in this study. The first 2 patients received both immediately loaded and submerged implants, while the remaining patients had all implants immediately loaded. The first 9 patients received a temporary prosthesis within 4 hours of surgery, and the hybrid prosthesis, made of a titanium framework and acrylic resin teeth, was placed after 6 months. The last 6 patients received the same type of hybrid prosthesis within 36 hours of surgery. Marginal bone loss was monitored via periapical radiographs by a computerized technique., Results: One failure (out of the 92 immediately loaded implants) occurred after 3 weeks of function because of infection. A cumulative success rate of 98.9% was achieved for up to 48 months of follow-up, while the prosthetic cumulative success rate for the same period was 100%. Marginal bone loss at the immediately loaded implants was within the generally accepted conventional limits for standard delayed loading protocols., Discussion: This technique can reduce treatment time but should be applied with caution., Conclusion: The preliminary results of this study suggest that rehabilitation of the edentulous mandible by an immediately loaded hybrid prosthesis supported by 5 to 6 implants may represent a viable alternative treatment to the classical delayed loading protocols.

This article describes two different surgical techniques of root coverage using Emdogain and shows preliminary results on 26 shallow recessions in 14 patients. For the treatment of 13 recessions, Emdogain was used in combination with a coronally advanced flap (CAF+EMD group). In the other 13 recessions, Emdogain and the flap were used in combination with a subepithelial connective tissue graft (CAF+CTG+EMD group). For the CAF+EMD group, the root coverage at 6 months was 93.97%, with an attachment gain of 3.2 mm; for the CAF+CTG+EMD group, the root coverage was 93.59%, with an attachment gain of 3.4 mm (no statistically significant difference between groups). When complete root coverage was not achieved, the residual recession was 1 mm in four cases and 2 mm in one case. Keratinized gingiva was increased for both groups, but more for the CAF+CTG+EMD group (1.38 mm versus 0.69 mm; statistically significant difference). Clinical attachment level decreased significantly in both groups, from 4.46 to 1.23 mm in the CAF+EMD group, and from 4.62 to 1.23 mm in the CAF+CTG+EMD group. Preliminary results show that Emdogain, in combination with CAF or CAF+CTG for the treatment of Miller Class I or II gingival recessions, displays good clinical results, with percentage of root coverage comparable or superior to other techniques. Further experimental studies on the dynamics of wound healing are needed to prove that EMD is really responsible for improving the percentage of regenerated versus repaired tissues with respect to other techniques.

A growing number of clinical studies show that early (2 months) and immediate loading protocols may be predictable. However, they are based on clinical stability only The aim of this case report was to document the osseointegration status of two Osseotite implants after 2 months of healing in soft bone corresponding to type IV and subjected to two distinct mechanical environments. A completely edentulous patient received a total of 11 Osseotite implants in the mandible. Six were immediately loaded to support a provisional fixed partial denture, and five were left submerged. After 2 months, two submerged and one immediately loaded implants were retrieved and processed for histologic analysis. All immediately loaded implants were clinically stable. One histologic section per implant was obtained. All implants achieved osseointegration. The bone-implant contact was 38.9% for the submerged implant and 64.2% for the immediately loaded one. In the marrow space, both implants were covered by thin, neoformed bone trabeculae. Osseointegration can be achieved after 2 months by Osseotite implants placed in soft bone in the mandible either when immediately loaded and splinted into a provisional denture retained on six implants, or when submerged and unloaded.

In this prospective study, the clinical outcome of 2-months loaded implants placed in the posterior jaws was evaluated after up to 3 years of functional loading. 405 Osseotite implants with microtextured acid-etched surface were consecutively placed in 11 fully and 164 partially edentulous patients (mean age 53.5 +/- 15.3 (S.D.) years) using a one-stage technique. 282 implants, supporting 154 restorations, were placed in the mandible, while 123 implants, supporting 75 restorations, were placed in the maxilla. A total of 99 single-tooth restorations, 119 short-span fixed bridges and 11 full-arch prostheses were applied. The mean time interval from surgery to connection of provisional restoration was 2.0 +/- 0.7 months. 9 failures were reported up to 3 years of follow-up; of these, 4 mandibular and 2 maxillary implants failed during the initial healing period (before prosthesis connection), while 3 mandibular implants failed after loading. No other complications occurred throughout the study. Life table analysis showed a cumulative survival rate of 97.5% for the mandible and 98.4% for the maxilla. Post-loading implant survival rate was 98.9% for the mandible and 100% for the maxilla. The excellent outcome of this interim report suggests that microtextured Osseotite implants in the posterior jaws may safely bear a functional load applied 2 months after insertion. The results of the present study are particularly encouraging for implants placed in the maxilla, since both a significant reduction of the healing period compared to the 6 months suggested by the classical Brånemark protocol, and an excellent post-loading function may be achieved also in low quality bone.

To study pulmonary arteriolar vasomotion in control conditions and in the transition to hydraulic edema, changes in subpleural pulmonary arteriolar diameter and perivascular interstitial volume were evaluated in anesthetized spontaneously breathing rabbits. Images of subpleural pulmonary microvessels were recorded in control conditions and for up to 180 min during a 0.5 ml x kg(-1) x min(-1) intravenous saline infusion through an intact parietal pleural window. Images were digitized and analyzed with a semiautomatic procedure to determine vessel diameter and perivascular interstitial thickness from which interstitial fluid volume was derived. In control vessels, the diameter of approximately 30-, approximately 50-, and approximately 80-microm arterioles and the perivascular interstitial thickness were fairly stable. During infusion, the diameter increased maximally by 20% in approximately 30 microm vessels, was unchanged in approximately 50 microm vessels, and decreased by 25% in approximately 80-microm arterioles; the perivascular interstitial volume increased by 54% only around 30-microm microvessels. In papaverine-treated rabbits, all arterioles dilated and a larger increase in perivascular interstitial thickness was observed. The data suggest that the opposite vasomotor behavior of 30- and 80-microm arterioles during development of mild edema may represent a local specific response of the pulmonary microcirculation to reduce capillary pressure in the face of an increased transendothelial fluid filtration, thus counteracting progression toward severe edema.

Both theoretical and experimental studies of pleural fluid dynamics and lung buoyancy during steady-state, apneic conditions are presented. The theory shows that steady-state, top-to-bottom pleural-liquid flow creates a pressure distribution that opposes lung buoyancy. These two forces may balance, permitting dynamic lung floating, but when they do not, pleural-pleural contact is required. The animal experiments examine pleural-liquid pressure distributions in response to simulated reduced gravity, achieved by lung inflation with perfluorocarbon liquid as compared to air. The resulting decrease in lung buoyancy modifies the force balance in the pleural fluid, which is reflected in its vertical pressure gradient. The data and model show that the decrease in buoyancy with perfluorocarbon inflation causes the vertical pressure gradient to approach hydrostatic. In the microgravity analogue, the pleural pressures would be toward a more uniform distribution, consistent with ventilation studies during space flight. The pleural liquid turnover predicted by the model is computed and found to be comparable to experimental values from the literature. The model provides the flow field, which can be used to develop a full transport theory for molecular and cellular constituents that are found in pleural fluid.

A growing number of clinical reports show that early and immediate loading of endosseous implants may lead to predictable osseointegration; however, these studies provide mostly short- to mid-term results based only on clinical mobility and radiographic observation. Other methods are needed to detect the possible presence of a thin fibrous interposition of tissue that could increase in the course of time and lead to clinical mobility A histologic evaluation was performed on two immediately loaded Osseotite implants retrieved after 4 months of function from one patient. He had received a total of 12 implants in the mandible, of which six were immediately loaded and six were left to heal in a submerged way Clinical and histologic osseointegration was consistently achieved for both of the retrieved immediately loaded implants. Osteogenesis and bone remodeling on the Osseotite surface were not impeded by immediate loading as shown by histomorphometric evaluation, which revealed high levels of bone-to-implant contact ranging from 78% to 85%. This immediate loading protocol involving bilateral splinting of six Osseotite implants in the mandible proved to be successful after 4 months of loading. Further long-term clinical and histologic studies are needed before introducing this immediate loading protocol as a routine procedure in implant therapy.

Pulmonary interstitial pressure was measured via micropuncture in anesthetized rabbits in normoxia and after breathing 12% O(2). In normoxia [arterial PO(2) = 88 +/- 2 (SD) mmHg], pulmonary arterial pressure and pulmonary interstitial pressure were 16 +/- 8 and -9.6 +/- 2 cmH(2)O, respectively. After 6 h of hypoxia (arterial PO(2) = 39 +/- 16 mm Hg), the corresponding values were 30+/-8 and 3.5+/-2.5 cm H(2)O (P<0.05). Pulmonary interstitial proteoglycan extractability, evaluated by hexuronate assay after 0.4 M guanidinium hydrochloride extraction, was 12.3, 32.4, and 60.6 microg/g wet tissue in normoxia and after 3 and 6 h of hypoxia, respectively, indicating a weakening of the noncovalent bonds linking proteoglycans to other extracellular matrix components. Gel filtration chromatography showed an increased fragmentation of chondroitin sulfate- and heparan sulfate-proteoglycans during hypoxic exposure, accounting for a loss of extracellular matrix native architecture and basement membrane structure. Gelatin zymography demonstrated increased amounts of the proteolytically activated form of gelatinase B (matrix metalloproteinase-9) after hypoxic exposure, providing evidence that the activation of proteinases may play a role in hypoxia-induced lung injury.

Background: An increase in gingival sulcular fluid filtration is a common clinical sign of early gingivitis. The aim of this study was to describe the fluid dynamics of the gingival interstitial tissues at the level of the sulcus in the transition towards inflammation., Methods: In 13 anesthetized rabbits, a silk ligature was placed around incisors close to the gingival margin, in order to prevent mechanical cleaning of plaque deposition. After 2 to 7 days, animals were anesthetized and interstitial fluid pressure measured using glass micropipettes connected to a servonull pressure system at the level of the free and attached gingiva., Results: Interstitial pressure was 3.8 +/- 2.9 cmH2O, significantly higher than the normal physiological value (about -1 cmH2O). Colloid osmotic pressure of interstitial fluid samples collected using the wick technique was measured using an osmometer whose membrane had a molecular cut-off of 30 kD and averaged 12.8 +/- 2.8 cmH2O (unchanged relative to control). Mean gingival sulcular fluid flow, measured by placing a PE tube (0.5 mm OD, 0.28 mm ID) in the sulcus, was 0.16 +/- 0.12 microl/h; the mean colloid osmotic pressure was 13.6 +/- 6.6 cmH2O, corresponding to a protein concentration of approximately 2.8 g/dl. Proteins of gingival fluid may leak from inflamed gingival interstitium or derive from bacteria of dental plaque. Histological analysis of gingival biopsies showed neutrophilic polymorphonuclear leukocyte infiltrates in both the dermis and epidermis layer., Conclusions: Based on hydraulic and colloid osmotic data, the Starling pressure gradient favored filtration from gingival interstitium to the sulcular space, a condition opposite to that observed in healthy gingiva where fluid filtration is absent.

An alteration of the immune system function is one of the main factors involved in the development of periodontal disease. Polymorpho-nuclear neutrophil leukocytes (PMN) play a crucial role in the cell-mediated immune response against bacterial challenge. The mechanism of neutralization of pathogen microorganisms by PMNs involves many different steps: adhesion to capillary endothelium in the inflamed region, trans-endothelial migration, chemotaxis, phagocytosis and, ultimately, bacterial killing by oxidative and non-oxidative mechanisms. A defect in one of these steps leads to altered neutrophil function and, consequently, to a higher host susceptibility to periodontal tissue infection. The main intrinsic neutrophil diseases such as neutropenia, leukocyte adhesion deficiency (LAD-1), Chediak-Higashi syndrome, Papillon-Lefèvre syndrome, chronic granulomatous disease (CGD), are often related to severe and early-onset forms of periodontitis, as described by many evidences in the literature. Therefore PMN dysfunctions, both intrinsic and extrinsic, represent an important risk factor for periodontal disease. Studies on the basic molecular mechanisms of such dysfunctions, also in terms of genetic polymorphisms, recently allowed to identify some specific markers related to a higher susceptibility to the development of disease. Many researches have yet to be performed aiming to gain insight on the dynamics of PMN activation and interaction with other cells, in order to improve and modulate neutrophil function and to develop specific approaches for care and prevention of periodontal diseases.

Background/aims: This clinical trial aimed at comparing 2 different means of delivering chlorhexidine digluconate (CHX) in the oral hygiene phase during the 2 weeks following periodontal surgery., Method: 40 patients were randomly divided into 2 groups: A (using CHX mouthwash) and B (using CHX spray). Professional oral hygiene was carried out immediately before surgical operation. For 7 days after surgery, group A used CHX mouthwash and group B CHX spray on the teeth involved in the surgical procedure, while mechanical oral hygiene was maintained only on the teeth not involved surgically. After suture removal, on the 7th day, patients were allowed to perform mechanical oral hygiene also on surgical sites. Plaque index (PI) and stain index (SI) were evaluated on the 7th and 14th days after the operation., Results: In both groups, PI increased similarly with respect to the baseline in surgically involved teeth, being 0.25+/-0.41 (SD) and 0.15+/-0.26, respectively, in A and B, on day 7, and 0.14+/-0.23 (A) and 0.10+/-0.22 (B), on day 14. There was no significant difference between A and B on either day 7 or 14. On the contrary, SI increased significantly in respect to the baseline over the 14 days in group A at both involved and not involved sites, while it did not differ from the baseline in group B., Conclusions: The present results indicate that the efficacy of CHX spray in the post-surgical control of dental plaque is not different from that of CHX mouthwash. Tooth staining, on the contrary, was significantly lower in the group using CHX spray. The observed effects might be related to the way of delivering CHX and to the total dose administered, about 80% lower in group B in respect to A. Further studies are needed to validate the preliminary findings of the present study.

Polymorphonuclear neutrophil granulocytes (PMN) are considered the most important cells of the host immune response against bacterial challenge. The functional mechanism of PMN consists of different steps: tethering, rolling, primary adhesion to the vascular wall, firm adhesion to the activated endothelium in the inflamed region, trans-migration across endothelium, chemotaxis, contact with the bacterium and phagocytosis and, finally, killing of the micro-organism by releasing hydrolytic enzymes and/or by production of toxic substances such as free radicals. Each of these steps is controlled by interactions between cells and many components of the immune system or inflammatory mediators. These interactions generate specific signals, important for cell regulation. Recent technological advances in molecular biology and immunobiology allowed to disclose the precise role of various molecules involved in the immune response, that regulate PMN function; conversely, more factors have been identified, whose role is still unknown. In the process of adhesion, for example, many classes of molecules are involved (selectins, integrins, ICAMs). The interaction of these molecules (es.: selectin) with their ligands (non completely discovered) is characteristic of specific stages, but may also regulate the successive steps (integrin activation). In periodontal infections, PMNs of gingival tissue migrate towards bacteria of dental plaque along a chemotactic gradient of specific factors (ICAM-1, IL-8) produced by cells of the junctional epithelium. Such gradient is essential to drive PMNs through molecular traffic. Among the mechanisms used by PMNs to kill bacteria, the importance of nitric oxide (NO) production has been recently pointed out.

1. Hydraulic pressure in intercostal and diaphragmatic lymphatic vessels was measured through the micropuncture technique in 23 anaesthetised paralysed rabbits. Pleural lymphatic vessels with diameters ranging from 55 to 950 microm were observed under stereomicroscope view about 3-4 h after intrapleural injection of 20 % fluorescent dextrans. 2. Lymphatic pressure oscillated from a minimum (Pmin) to a maximum (Pmax) value, reflecting oscillations in phase with cardiac activity (cardiogenic oscillations) and lymphatic myogenic activity. With intact pleural space, Pmin in submesothelial diaphragmatic lymphatic vessels of the lateral apposition zone was -9.1 +/- 4.2 mmHg, more subatmospheric than the simultaneously recorded pleural liquid pressure amounting to -3.9 +/- 1.2 mmHg. In extrapleural intercostal lymphatic vessels Pmin averaged -1.3 +/- 2. 7 mmHg. 3. Cardiogenic pressure oscillations (Pmax - Pmin), were observed in all recordings; their mean amplitude was about 5 mmHg and was not dependent upon frequency of cardiac contraction, nor lymphatic vessel diameter, nor the Pmin value. 4. Intrinsic contractions of lymphatic vessel walls caused spontaneous pressure waves of about 7 mmHg in amplitude at a rate of 8 cycles min-1. 5. These results demonstrated the ability of pleural lymphatic vessels to generate pressure oscillations driving fluid from the subatmospheric pleural space into the lymphatic network.

Gingival hydraulic interstitial pressure was measured with glass micropipettes in 18 anesthetized rabbits at the level of the free gingiva, attached gingiva and oral mucosa facing the incisors and molar teeth. Samples of gingival interstitial tissue fluid were also collected by inserting nylon wicks in the subepithelial layer of the oral mucosa. Colloid osmotic pressure of interstitial fluid samples was measured with an osmometer whose membrane had a molecular cutoff of 30 kD. Hydraulic interstitial pressure from the free gingiva, at an average distance of 300 microns from sulcular space, was -1.3 +/- 0.9 (SD) cmH2O. Mean colloid osmotic pressure of gingival tissues interstitial fluid was 13.1 cmH2O, corresponding to a protein concentration of 2.8 g/dl. The thickness of the sulcular epithelium and of the oral gingival epithelium (data from 2 rabbits) were approximately 100 microns; the minimal distance of microvessels from the surface of the sulcular epithelium was approximately 150 microns. Based on hydraulic and colloid osmotic data, the Starling balance of pressures causes fluid filtration from gingival capillaries to gingival interstitium; however, across the sulcular epithelium, the pressure gradient sustains fluid absorption from the sulcus into the gingival interstitium. Plasma proteins may leak from microvessels into gingival interstitium, down convective bulk flow and via a concentration dependent diffusive component. At sulcular level, proteins may leak into the sulcus down a concentration gradient. Thus, at sulcular level a peculiar condition occurs in that there is an absorption gradient for water but a filtration gradient for plasma proteins.

Knowledge of pleural liquid pressure (Pliq) and composition is crucial for studies concerning intrapleural fluid dynamics, and pleural fluid turnover. We measured Pliq at intercostal and costal levels in anaesthetized spontaneously breathing rabbits using a minimally invasive method that assures a long-lasting hydraulic continuity between the pleural liquid and the recording system. Polyethylene tubes were glued either to the exposed endothoracic fascia or inserted into a rib to provide a scaled connection to the recording system. After inducing a pneumothorax with nitrous oxide (N2O) via an intrapleural cannula, a hole (approximately 0.7 mm2) was pierced in the parietal pleura through the tube lumen. The tubes were then connected to pressure transducers and the whole system was filled with heparinized saline to the level of the parietal pleura; finally the pneumo-thorax was removed after N2O washout and Pliq recordings were performed. A different kind of tube was used to obtain microsamples of pleural fluid (2.5-3 microliters) during spontaneous breathing; colloid osmotic pressure of the microsamples (pi liq) was measured with an osmometer, and averaged 9.3 +/- 1.5 cm H2o (n = 70 samples). When pooled and plotted against lung height end-expiratory intercostal and costal Pliq data scattered along a single regression line with a slope of -0.83 and -0.90 cm H2O cm(-1) in supine and prone animals, respectively. End-inspiratory costal Pliq was significantly more subatmospheric than intercostal in the ventral region of the chest (P < 0.05), and less subatmospheric in the dorsal region, regardless of posture. The techniques presented here could be helpful in gaining a greater insight into the physiology and pathophysiology of the pleural space in terms of pleural fluid dynamics and turnover.

Objective: To develop an algorithm to detect the edges between lung tissue, perivascular interstitium, and microvessel using digital processing of in vivo microscopic images of lung surface., Methods: A numerical technique was developed to identify three different regions (namely, pulmonary microvessel, perivascular interstitium, and lung tissue) based on their corresponding gray level distributions. We present a theoretical demonstration of the method and a semiautomatic procedure that, once the edges are detected, determines microvascular diameters and perivascular interstitium thickness., Results: Microvessel diameters and perivascular interstitium thickness were calculated for precapillary arteriolar branching (40 to 140 microns) and saved in an ASCII file., Conclusions: We proved that the maximum value of the moving variance is useful to detect the edge between two adjacent regions whose gray level distributions satisfy the condition: magnitude of sigma Y2 - sigma X2 < or = (mu X - mu Y)2, where mu X, mu Y, sigma X2, sigma Y2 are the statistical moments of the two regions X and Y. Moreover, when the regions have similar means, the above conditions is not met, but the edge between them can be detected by the maximum of the moving variance error.

By micropuncture technique we measured pulmonary interstitial pressure (Pip) from birth up to 6 h postnatal age in anesthetized and paralyzed cesarian delivered term (31 days gestation) and premature (27 to 30 days gestation) rabbits. In term cesarian delivered rabbits Pip followed the time course of vaginally delivered rabbits, namely, it increased from about zero at birth up to about 5 cmH2O at 2 h, as a result of alveolar fluid reabsorption, subsequently it decreased becoming subatmospheric due to progressive interstitial fluid drainage. In ventilated lung regions of premature rabbits, Pip also peaked to about 5 cmH2O at 2 h but its subsequent decrease was markedly slowed down while in atelectatic regions of premature rabbits Pip remained slightly subatmospheric. Up to 6 h, the wet/dry weight ratio of the lung was higher in premature relative to vaginally and cesarian delivered term rabbits (at birth 8.4 +/- 0.9 vs. 7.5 +/- 0.8). In 29-31 days rabbits, plasma protein concentration at birth was 3.6 +/- 0.5 g/dl (within 95% confidence limits for vaginally delivered rabbits, considered as control) while in 27-28 days rabbit it was 3.1 +/- 0.4 g/dl (at the lower edge of control confidence limits). In the first postnatal hours, the increase in Pip favoured fluid reabsorption into pulmonary microcirculation in term cesarian delivered rabbits and in ventilated regions of premature rabbits. Conversely, in the atelectatic regions of premature rabbits the unchanged Pip value in the postnatal hours favours fluid filtration from microcirculation into lung interstitium.

Objective: To determine microvascular diameter and perivascular interstitium thickness at the lung surface in the in situ, in vivo lung., Methods: Microscopic images of the lung surface collected through a "pleural window" by a videocamera were digitized with a monochrome frame grabber (512 x 512 pixels, 8 bits per pixels) to be computer analyzed by image processing techniques., Results: We found that the maxima in the distribution of the standard deviations of gray levels in adjacent neighbors 7 x 7 pixels wide identify the edges between the microvessel lumen and the surrounding perivascular interstitium. Furthermore, the maxima in the distribution of the standard deviation of the standard deviations of gray levels identify the edges between the perivascular interstitium and the lung tissue., Conclusions: This technique can be applied to microvessels ranging in diameter from 30 microns to 200 microns and perivascular interstitial thickness of the order of 10-150 microns. Our approach allows for the definition of microvascular geometry even for noisy images and represents an improvement compared to other edge detection methods. The proposed analytical procedure may provide a useful tool to study lung fluid balance and microvascular reactivity in the in situ lung in the normal state and in response to a variety of functional conditions.

In anesthetized paralyzed term newborn rabbits at various postgestational ages (from birth up to 16 days), we measured by micropuncture technique the hydraulic pressure of the pulmonary interstitium (Pip), the extrapleural parietal interstitium, and the pleural liquid. Birth data refer to cesarian-delivered nonbreathing rabbits. Pip increased from 0.5 +/- 2 to 6 +/- 0.7 cmH2O from birth up to 2 h and then decreased, becoming subatmospheric at 5 h and attaining -6 +/- 1.6 cmH2O at 16 days. Over the same period of time, pressure in the extrapleural parietal interstitium and the pleural liquid remained fairly constant at an average value of approximately -1.5 and -2 cmH2O, respectively. The wet-to-dry weight ratio of the lungs decreased from 7.8 +/- 0.4 to 4.9 +/- 0.1 at 16 days. Plasma protein concentration was 4.2 +/- 0.4 g/dl at birth, decreased to 3.2 +/- 0.5 g/dl at 1 h from delivery, and increased back to 4 +/- 0.6 g/dl at 16 days. Pleural liquid protein concentration was 3 +/- 0.1 g/dl at birth and decreased to 1.2 +/- 0.2 g/dl at 16 days. In the first hours of postnatal life, the marked increase in Pip appears to be a key factor in favoring fluid clearance from pulmonary interstitium into the pulmonary capillaries and the pleural space. This factor vanishes after approximately 6 h because of the marked decrease in Pip.

A model of pleural fluid turnover, based on mass conservation law, was developed from experimental evidence that 1) pleural fluid filters through the parietal pleura and is drained by parietal lymphatics and 2) lymph flow increases after an increase in pleural liquid volume, attaining a maximum value 10 times greater than control. From the differential equation describing the time evolution of pleural liquid pressure, we obtained the equation for the steady-state condition ("set point") of pleural liquid pressure: Pss = (KfPi*+KlPzf)/Kf+Kl), where Kf is parietal pleura filtration coefficient, Kl is initial lymphatic conductance, Pzf is lymphatic potential absorption pressure, and Pi* is a factor accounting for the protein reflection coefficient of parietal mesothelium and hydraulic and colloid osmotic pressure of parietal interstitium and pleural liquid. Lymphatics act as a passive negative-feedback control tending to offset increases in pleural liquid volume. Some features of this control are summarized here: 1) lymphatics exert a tight control on pleural liquid volume or pressure so that the set point is maintained close to the potential absorption pressure of lymphatics; 2) a 10-fold increase in Kf would cause only a 2- and 5-fold increase in pleural liquid volume with normal (1.8 g/dl) and increased (3.4 g/dl) protein concentration of the pleural fluid, respectively; and 3) the reduction in maximum lymph flow greatly reduces the range of operation of the control with increased filtration and/or protein concentration of pleural fluid.

In 31 anesthetized rabbits, after removal of superficial tissues, glass micropipettes filled with 0.5 M NaCl solution and connected to an electrohydraulic servo-null system were used to measure extraperitoneal interstitial fluid pressure (Pi,per) and peritoneal liquid pressure (Pliq,per) at various heights. Linear regressions relating pressure to recording height (H) were Pi,per = 1.07 - 0.27H and Pliq,per = 0.9 - 0.64H, respectively. Protein concentration (Cp;g/dl) and colloid osmotic pressure (II; cmH2O) of plasma and of peritoneal and pleural liquids were 5.48 +/- 0.38 and 24.61 +/- 3.23, 3.07 +/- 0.5 and 13.29 +/- 1.87, and 1.76 +/- 0.42 and 8.45 +/- 2, respectively. The equation relating II to Cp was II = 4.64Cp + 0.0027Cp2. Tissue fluid samples were collected with saline-soaked wicks implanted in vivo or dry wicks inserted postmortem in extraperitoneal and extrapleural interstitial spaces. After 60 and 15 min, respectively, wicks were withdrawn and centrifuged; wick fluid was analyzed in colloid osmometer for small samples. Average extraperitoneal and extrapleural II values were 14.2 +/- 2.49 and 11.94 +/- 1.52 cmH2O, corresponding to Cp of 3.07 and 2.57 g/dl, respectively. The average net pressure gradient, assuming reflection coefficient and hydraulic conductivity (Negrini et al. J. Appl. Physiol. 69: 625-630, 1990; 71: 2543-2547, 1991), was 1.18 and 0.98 cmH2O for parietal peritoneal and pleural mesothelia, respectively, favoring filtration from the extraserosal interstitia into the serosal cavities. Total parietal peritoneal filtration was 1.49 ml.kg-1.h-1, approximately 15-fold higher than that for pleural mesothelium.

In anesthetized rabbits (n = 25) subject to slow intravenous saline loading (0.4 ml.min-1.kg-1) for 3 h, we measured pulmonary interstitial pressure (Pip) in intact in situ lungs with glass micropipettes inserted directly into the lung parenchyma via a "pleural window." Measurements were done in apneic animals at the end-expiratory volume with O2 delivered in the trachea. Pip was -10 +/- 1.5 (SD) cmH2O in control and increased to 0.6 +/- 3.8 and 5.7 +/- 3.3 cmH2O at 66 and 180 min, respectively. The wet-to-dry weight ratio (W/D) of the lung was 5.04 +/- 0.2 in the control group and 5.34 +/- 0.7 at 180 min (+6%); the corresponding W/D for intercostal muscles were 3.25 +/- 0.03 and 4.19 +/- 0.5 (+28%). Pulmonary interstitial compliance was 0.47 ml.mmHg-1.100 g wet wt-1. Pulmonary arterial and left atrial pressures were 18.4 +/- 2 and 3 +/- 1 cmH2O in control and increased to 19.5 +/- 2.9 and 4.6 +/- 1.7 cmH2O at 180 min, respectively. Aortic flow (cardiac output) increased from 103 +/- 35 to 131 +/- 26 ml/min; pulmonary resistance fell from 0.17 +/- 0.06 to 0.14 +/- 0.05 cmH2O.min.ml-1 (-18%), suggesting that the increase in Pip did not limit blood flow. The pulmonary capillary-to-interstitium filtration pressure gradient decreased sharply from a control value of 10 cmH2O to 0 cmH2O within 60 min because of the increase in Pip and remained unchanged for < or = 180 min. Data suggest that the pulmonary interstitial matrix can withstand fluid pressures above atmospheric, preventing the development of pulmonary alveolar flooding.

The morphology of the submesothelial lymphatic lacunae on the pleural and peritoneal surface over the tendinous and muscular portion of the diaphragm was studied in 10 anesthetized rabbits. The lymphatic network was evidenced by injecting 1 ml of colloidal carbon solution in the pleural (n = 5) or the peritoneal (n = 5) space. After 1 h of spontaneous breathing, the animal was killed and the diaphragm was fixed in situ by injection of approximately 5 ml of fixative in pleural and peritoneal spaces. Then both cavities were opened and the diaphragm was excised and pinned to a support. According to which cavity had received the injection, the peritoneal or the pleural side of the diaphragm was scanned by sequential imaging of the whole surface by use of a video camera connected to a stereomicroscope and to a video monitor. The anatomic design appeared as a network of lacunae running either parallel or perpendicular to the major axis of the tendinous or muscular fibers. The lacunae were more densely distributed on the tendinous peritoneal area than on the pleural one. Scanty lacunae were seen on the muscular regions of both diaphragmatic sides, characterized by large areas without lacunae. The average density of lacunae on tendinous and muscular regions was 6 and 1.7/cm2 for the pleural side and 25 and 3.4/cm2 for the peritoneal side, respectively. The average width of lacunae was 137.9 +/- 1.6 and 108.8 +/- 1.7 microns on the tendinous pleural and the peritoneal side, respectively, and 163 +/- 1.8 microns on the muscular portion of the pleural and peritoneal surfaces.

In seven anesthetized rabbits we measured the size, shape, and density of lymphatic stomata on the peritoneal and pleural sides of the diaphragm. The diaphragm was fixed in situ and processed for scanning electron microscopy. Results are from 2,902 peritoneal and 3,086 pleural fields (each 1,620 microns 2) randomly chosen from the various specimens. Stomata were seen in 9% of the fields examined, and in 30% of the cases they appeared grouped in clusters with 2-14 stomata/field. Stoma density was 250 +/- 242 and 72 +/- 57 (SD) stomata/mm2 on peritoneal and pleural sides, respectively, and it was similar over the muscular and tendinous portion of the two surfaces. The maximum diameter ranged from less than 1 to approximately 30 microns, with an average value of 1.2 +/- 3.1 micron. The ratio of the maximum to the minimum diameter and the surface area averaged 2 +/- 1.4 and 0.7 +/- 2.4 micron 2, respectively. The maximum and minimum diameter and surface area values followed a lognormal frequency distribution, suggesting that stomata geometry is affected by diaphragmatic tension.

The peritoneal cavity of 18 anesthetized spontaneously breathing supine rabbits was opened through a midline section. One or two hollow capsules (surface area 0.8 cm2) were glued to the exposed liver surface, filled with whole or 25% diluted plasma, and connected to a transducer and a graduated pipette. Various hydraulic pressures (Pcap) were set in the capsule; at each Pcap the liquid flow per unit surface area (V/S) between the Disse's interstitial space and the capsule was measured from the rate of liquid displacement in the pipette. The slope of the V/S vs. Pcap linear regression was utilized to estimate the hydraulic conductivity of the Glissonian-peritoneal membrane and averaged 5.1 x 10(-3) +/- 4.7 x 10(-3) (SD) ml.h-1.cmH2O-1.cm-2 (n = 25). Hydraulic pressure in the Disse's space (Pd) was measured by closing the capsule against the transducer disconnected from the pipette. At portal and hepatic venous pressures of 7.6 +/- 2.9 and 2.6 +/- 1 cmH2O, respectively, Pd was 2.05 +/- 2 cmH2O. Physiologically, Starling pressure gradients cause fluid transfer from the sinusoids to the Disse's space; transperitoneal fluid filtration only occurs through the liver surface that faces the diaphragm, which corresponds to one-fifth of the total hepatic surface.

The macroagglutination test, according Mailloux, was investigated for its feasibility in the rapid diagnosis of human and animal leptospirosis. Suspected sera examinated by Mailloux test, were also examinated by Complement Fixation and Microagglutination; the results suggest that: Mailloux macroagglutination is the serological test of choice, for screening of animal and human sera, mostly if it is not needed to know the infecting serovar.

Data from studies of the metabolic mechanism of the sodium dependence of halophilic leptospirae revealed that the presence of sodium - 1) has no effect on the oxygen uptake when the microorganism is oxidizing endogeneous and exogeneous substrates; 2) is not involved in the multiplication phase; but 3) is of importance for keeping the cell structure intact.

In 14 spontaneously breathing anesthetized rabbits, we used cyanoacrylate to glue a hollow capsule, at end expiration or at end inspiration, to the peritoneal surface of the tendinous portion of the diaphragm. The capsule was connected to a pressure transducer and a pipette calibrated in microliters. We filled the system with fluid and measured flow into the diaphragmatic surface facing the capsule (Fcap, microliter/cm2), from liquid displacement in the pipette at different hydraulic pressures in the system (Pcap). Pleural liquid pressure was simultaneously measured in the supraphrenic region (Psup). Fcap was positively correlated to transdiaphragmatic pressure gradient (Psup-Pcap) and breathing frequency but was unaffected by protein concentration of capsular fluid. For a breathing frequency of 30 cycles/min and a Psup - Pcap = -2 cmH2O, Fcap was 0.54 microliter.min-1.cm-2 for capsules applied at end expiration and 10-fold greater for capsules applied at end inspiration. Data indicate that the diaphragmatic tendinous portion in rabbits is a draining site for peritoneal fluid and that the conductance of the draining pathways (lymphatic stomata) is related to diaphragmatic tension. In the intact rabbit the average peritoneal fluid drainage through the tendinous portion of the diaphragm (approximately 16 cm2) was estimated at 43 microliters/min.