[The impact of periodontal disease treatment on endothelium of sublingual microvessels]. / Vliyanie khirurgicheskogo lecheniya parodontita na sostoyanie endoteliya mikrososudov pod''yazychnoi oblasti.

The intactness degree of glycocalyx and the thickness of endothelium layer in sublingual microvessels was determined by means of darkfield microscopy and GlycoCheck RI software in 35 patients (19 men and 16 women, aged 35 to 65 years) with chronic generalized severe periodontitis before and after anti-inflammatory and surgical treatment. The patients were divided into two groups. The first group recieved only local anti-inflammatory therapy while patients of group 2 underwent surgical intervention on the periodontium in addition to local anti-inflammatory therapy. In both groups the changes of parameters of endothelial glycocalyx (EGC) in the early stages of treatment were not significant. In long terms the changes were not revealed in group 1, but in group 2 6 months after surgical treatment in all patients the decrease in the thickness of the permeable glycocalyx decreased and increase of the volume of red blood cells filling was observed.

Measurement of single cell refractive index, dry mass, volume, and density using a transillumination microscope.

Phase contrast microscopy has become ubiquitous in the field of biology, particularly in qualitative investigations of cellular morphology. However, the use of quantitative phase retrieval methods and their connection to cellular refractive index and dry mass density remain under utilized. This is due in part to the restriction of phase and cellular mass determination to custom built instruments, involved mathematical analysis, and prohibitive sample perturbations. We introduce tomographic bright field imaging, an accessible optical imaging technique enabling the three dimensional measurement of cellular refractive index and dry mass density using a standard transillumination optical microscope. The validity of the technique is demonstrated on polystyrene spheres. The technique is then applied to the measurement of the refractive index, dry mass, volume, and density of red blood cells. This optical technique enables a simple and robust means to perform quantitative investigations of engineered and biological specimens in three dimensions using standard optical microscopes.

Predictive factors for anemia within the first year after orthotopic liver transplantation.

We sought to determine the prevalence and predictive factors for posttransplant anemia within the first year after orthotopic liver transplant (OLT) among 97 consecutive patients. Anemia was defined at months 6 and 12 according to the WHO criteria, that is, a hemoglobin (Hb) level of <12 g/dL for women and <13 g/dL for men. Immunosuppression relied on tacrolimus and steroids, with or without mycophenolate mofetil. Anemia was present in 64.5%, 50%, and 52.8% of patients pre-OLT versus 6 and 12. Thirty-three percent (month 6) and 30.3% (month 12) of anemic patients received recombinant erythropoietin therapy. A multivariate analysis revealed that the independent predictive factors for anemia at month 6 were mean corpuscular volume (<85 fL) at day 7, daily steroid dosage (<0.3 mg/kg), serum creatinine (>130 mumol/L), and Hb level (<11 g/dL) at month 1. Independent predictive factors for anemia at month 12 were daily steroid dosage at month 1 (<0.3 mg/kg), hematocrit at month 1 (<33%), red blood cell count at month 6 (<3.75 T/L), daily dosage at month 1 of cyclosporine or tacrolimus, and etiology of end-stage liver disease other than alcohol abuse. We concluded that anemia was highly prevalent within the first year of post-OLT. This observation deserves further investigation and appropriate treatment.

Experiments on extracted teeth into the validity of using laser Doppler techniques for recording pulpal blood flow.

The performances of two laser Doppler flow meters (Periflux model PF3 and Moor Blood Flow Monitor model MBF3D) were investigated. Recordings were made while diluted blood was pumped at different rates through the pulp cavities of extracted human or pig teeth. The probe of each instrument was fixed to the enamel surface 2 mm from the original position of the gingival margin. Both instruments performed similarly, although the Moor gave the better signal/noise ratios. Both were capable of detecting flow in the core of the pulp as well as the superficial part closest to the probe tip. With either instrument, and with any one dilution of blood (range 0.5-45% v/v red cells), there was a near-linear relationship between the blood-flow signal and the flow rate of blood through the tooth. However, when data obtained with different red-cell concentrations were compared, a good correlation between the blood-flow signal and red-cell flux (product of concentration and mean velocity) was obtained only with red-cell concentrations up to 1% v/v. Therefore these instruments would give an accurate indication of blood flow changes only under conditions in which either this value was not exceeded or if the red-cell volume fraction remained constant; neither of which can be assumed to apply when recording from teeth in situ. The signals representing the concentration of moving blood cells were unreliable in both instruments. It is concluded that the information provided by these laser Doppler flow meters can be ambiguous and must be interpreted with care.

Pressure-flow relationship of erythrocyte suspension in perfusion of nuclepore membrane and red cell deformability.

The hemodynamic characteristics of Nuclepore (NP) membrane filtration were evaluated from a pressure (P)-flow rate (Q) relationship of erythrocyte suspension obtained by the vertical-tube method (Nichol et al., 1951) with a slight modification. It became evident that the vertical-tube method was a more quantitative and simple method than the conventional steady flow perfusion method using a variable speed pump. The P-Q relationship of erythrocyte suspensions in the perfusion of NP membrane consisted of a smooth curve convex to the P-axis at low P tending to a straight line at high P. The extrapolated linear segments of P-Q curves of erythrocyte suspension did not converge to the same point on the negative Q-axis, that is, the value of the negative intercept increased as flow decreased. This observation indicated clearly an obstruction or plugging of the pores by erythrocytes. It was made clear that flow rate measured with NP membrane filtration was influenced not only by intrinsic red cell deformability but also by several different factors, namely, the different distribution of pore size of NP membrane, microdust contaminated inevitably in red cell suspension, total volume of cell suspension through NP membrane, stagnation or retention of erythrocyte suspension and the hematocrit value of suspension. In conclusion, the present study shows that the precise definition of deformability depends on the method or rheological conditions, such as hematocrit value of suspension, used to measure it, even if confined within NP membrane filtration.

Research on oxygen transport in jaw cysts: investigation of changes of erythrocyte membrane fluidity.

I have investigated oxygen transport in the fluid from jaw cysts. Measurements were made of erythrocyte membrane fluidity, mean corpuscular volume, PO2, leucocyte percentage and erythrocyte and leucocyte numbers, both before and after irrigation of the cysts with antibiotics. On the basis of the results it is suggested that the membrane fluidity of erythrocytes in jaw cysts is decreased by infection and the PO2 value falls as a result of this decrease and also because of the increase in mean corpuscular volume and the raised oxygen consumption due to neutrophilia within jaw cysts.

Biofiltration vs. bicarbonate dialysis: influence on plasma volume changes and extravascular fluid mobilization.

The present study compares the effects of bicarbonate hemodialysis (Bic. HD) and biofiltration (BF), a new hemodiafiltration technique, on plasma volume (PV) changes and extravascular fluid mobilization (Vfm). Ten uremic patients underwent one experimental session of Bic. HD and, one week later, one of BF, both on the second dialysis of the week. Net ultrafiltration rate was limited to 700 ml/min. At the start of each session, whole blood volume (WBV), PV and red cell volume (RCV) were determined using 5 mu Ci of radioiodinated serum albumin (RISA). PV and Vfm were calculated at hourly intervals using a serial hematocrit method. On Bic. HD, PV increased at 60 min. then decreased at 120 and 180 min., with efficient Vfm only during the first hour. On BF, PV increased throughout treatment, with greater Vfm. It would appear that PV is better preserved in BF, on account of more efficient Vfm.

Blood profile and histology in oral infections associated with diabetes.

INTRODUCTION: We aimed to investigate the relationship between blood profile and histologic findings in both apical periodontitis (AP) and periodontal disease (PD) associated with diabetes. METHODS: Wistar rats (N = 80) were assigned to the following 8 groups: control, AP, PD, AP associated with PD, diabetes, diabetes with AP, diabetes with PD, and diabetes with AP and PD. Diabetes mellitus (DM) was induced with streptozotocin, AP was induced by exposure to the oral environment, and PD was induced using periodontal ligature. After 30 days, blood samples were collected, and the rats were euthanized. Subsequently, the maxillae were processed for light microscopy. Hematologic examinations were conducted to determine the total number of erythrocytes and leukocytes, erythrocyte constant, and blood glucose level. One-way analysis of variance and Kruskal-Wallis tests were used for statistical analysis, and the significance was set at P < .05. RESULTS: A significant correlation was found between the histologic findings and blood parameters. CONCLUSIONS: In conclusion, diabetes accelerated the development and progression of AP and PD in the rats and caused an increase in the average erythrocyte volume as well as the leukocyte and neutrophil counts. Oral infections increase the total number of leukocytes, the number of neutrophils and lymphocytes, and blood glucose concentrations in DM rats.

Effects of epinephrine on lidocaine pharmacokinetics and blood volume in the dental pulp.

INTRODUCTION: Epinephrine potentiates and prolongs the efficacy of local anesthetics by reducing blood flow. We investigated the effect of epinephrine on the pharmacokinetics of lidocaine and the pulpal blood volume after maxillary infiltration anesthesia in rats. METHODS: We measured the (14)C-radioactivity and (14)C-distribution in the maxilla and the dental pulp after the injection of 2% (14)C-lidocaine with or without 10 µg/mL epinephrine (n = 7) into the palatine mucosa proximal to the first molar. The blood volume in the pulp was measured using (99m)Tc-pertechnetate (n = 5). RESULTS: When lidocaine was injected together with epinephrine, the lidocaine became widely distributed throughout the maxilla and was observed mainly in the first molar pulp. The lidocaine amount in the dental pulp at 10-60 minutes was more than 2 times higher than that after the injection of lidocaine alone. The relative pulpal blood volume after 20 minutes decreased to 63.1% of the value after the injection of lidocaine alone. CONCLUSIONS: We found that lidocaine had infiltrated into the molar pulp after infiltration anesthesia. Furthermore, our results suggested that epinephrine augmented the retention of lidocaine in the pulp.

[Influence of heat-induced changes in the mechanical properties of the membrane on the filterability of human erythrocytes]. / Einfluss thermisch induzierter Veränderungen mechanischer Membraneigenschaften auf die Filtrierbarkeit menschlicher Erythrozyten.

The influence of heat induced changes of mechanical membrane properties of human erythrocytes on their filterability was investigated. A 10-minute incubation at 47, 48 and 48.4 degrees C, respectively, leads to an increase of elastic membrane shear modulus (up to 130%, 240%, 260%) and membrane viscosity (up to 220%, 380%, 480%). In comparison to these effects the filterability determined by cellulose filters was diminished relatively slightly (increase of filterability index up to 110%, 130%, 170%), with the mean erythrocyte volume remaining unchanged. Taking into account former investigations it is concluded that the used filtration technique is less sensitive with respect to changes of the elastic and viscous membrane properties compared with changes of cytoplasmic viscosity of the cells.

Measurements on the forward light scattering intensities of monodisperse latex particles and sphered or unsphered red blood cells.

Characteristics with respect to light scattering intensity (LSI) were measured on latex spheres and on sphered and unsphered red blood cells (RBC). From the discrepancies in LSI signals from polymer spheres and RBCs it is obvious that latices cannot be used for absolute calibration of RBC sizes. Sphering of RBCs did not give rise to a closer resemblance. Monodisperse latex spheres produce extremely narrow size distribution histograms. Regarding these sharp peaks and the long-term stability, polymer spheres offer definite advantages if compared with blood controls to which preservatives were added. A shift from the RBC size distribution histogram, caused by gradually developing instrument drift, can be easily detected and corrected at an early stage.

Surrogate materials for calibration and control: the use of latex particles as calibrants for red cell volume measurements.

Latex particles, including BCR Certified Reference Material CRM 166a, have important applications for checking linearity and for calibrating aperture-impedance instruments used to determine red-cell volumes. Studies performed on Coulter Counters, e.g. the ZBi/C1000, ZM/C256 and Model S Plus IV, using a series of three different mono-sized latex particles have shown linearity but non-zero intercepts. This emphasizes the importance of calibrating instruments at more than one point. Latex particles also have an important role in calibration of red-cell volume measurements provided that a relative latex: red-cell shape factor has been assigned when the instrument is calibrated with fresh blood. Such relative shape factors are independent of latex particle size but need to be specific for instrument type and probably also for the orifice and diluent to be used. Nevertheless latex calibrants can serve an important role in standardizing red-cell volume measurements.

The measurement of lymphocyte volume: importance of reference particle deformability and counting solution tonicity.

We have determined the influence of reference particle deformability and suspending buffer tonicity on the measurement of lymphocyte volume by an electronic particle volume analyzer. When the volume analyzer was standardized with latex spherules having a shape factor (fe) of 1.5, red cell volume was 96 cu micron and lymphocyte volume was 289 cu micron. The red cell volume corresponded closely to the true red cell volume; the true lymphocyte volume, however, was 218 cu micron when measured by the lymphocytocrit/lymphocyte count and 203 cu micron by wet lymphocyte weight and density (mean approximately 210 cu micron). The difference between the electronic volume (Ve) of 289 cu micron and true lymphocyte volume of 210 cu micron was due to the influence of lymphocyte deformability (shape factor) as it traverses the sizing aperture. Since the true volume equals the Ve/fe, the red cells with a shape factor near 1.0 were sized appropriately by this method. In contrast, the lymphocyte shape factor was 1.38; thus, the true lymphocyte volume was 289 cu micron/1.38 or 210 cu micron. The tonicity of the suspending solution also influenced the measurement of particle volume when osmotically inactive standard particles (e.g., latex spherules) were used as a reference. Whereas the true lymphocyte volume was 210 cu micron at 286 mosmole/liter, it was 194 cu micron at 330 and 229 cu micron at 250 mosmole/liter. The standard counting solution, Isoton, is hyperosmolar (330 mosmole/liter) and causes an 8% shrinkage of osmotically active cells.

A new cellulose acetate filter to remove leukocytes from buffy-coat-poor red cell concentrates.

Transfusion of leukocyte-free red cell concentrates (RCC) prevents or delays HLA immunization in multitransfused patients. We investigated a new cellulose acetate filter which was recently introduced to remove leukocytes from buffy-coat-poor RCC. It was found that the filtration time was only 10 min with buffy-coat-poor RCC in saline-adenine-glucose-mannitol (SAG M; n = 23), hematocrit being 62 +/- 2% (SD). The red cell loss was 13.5 +/- 2.6% and leukocyte removal was more than 99%. Routine filtration in SAG M (n = 179) showed again that more than 99% of the leukocytes were removed from buffy-coat-poor RCC with an original leukocyte content of 804 +/- (SD)458 x 10(6). The red cell loss (12 +/- 8.6%) was not diminished by increasing the amount of saline (0.9% NaCl) from 100 to 300 ml in an attempt to remove the retained red cells from the filter. We conclude that the new filter is reliable in rapidly removing more than 99% of the leukocytes from buffy-coat-poor RCC in SAG M solution.

The effect of chronic red cell mass depletion on the healing of bone grafts.

The purpose of this study was to determine whether a marked decrease in red cell mass with a normal blood volume (normovolemic anemia) would compromise osseous repair in experimental mandibular bone grafts. In adult male dogs, osseous defects in the mandibular body were restored with combined allogeneic and autologous bone grafts. In the experimental group, the red cell mass was depleted by a mean 55% of the baseline value and the resulting anemia maintained for eight weeks by serial phlebotomies. In the control group phlebotomy was not performed. Osseous repair was evaluated by quantitative sequential radionuclide imaging, subjective and quantitative determinations of graft stability, and histologic examination. Results demonstrated no significant difference in bone graft repairs between the anemic and control animals by any of the evaluation methods over the eight-week period of study (P greater than 0.05).

Effect of graft material on loss of erythrocytes after aortic operations.

It has been suggested that loss of erythrocytes after abdominal aortic grafting is influenced by the type of synthetic graft used. A prospective randomized study was done to compare loss of erythrocytes in patients receiving Dacron (polyester fiber, Meadox woven double velour) and Gore-Tex (polytetrafluoroethylene [PTFE]) grafts during the perioperative period. A total of 25 patients (13 Dacron and 12 PTFE) was studied, including 21 with abdominal aortic aneurysms and four with aortoiliac occlusive disease. Erythrocyte volume (EV) was measured using 51Cr-labeled autologous erythrocytes on the day prior to the operation, one to two hours after the operation when the patients were hemodynamically stable and 24 hours postoperatively. In addition to measurements of 51Cr EV and the volume of intraoperatively salvaged washed erythrocytes, the length of storage of the units of homologous liquid preserved erythrocytes at 4 degrees C. prior to transfusion were recorded. The mean intraoperative erythrocyte loss (+/- S.D.) for the Dacron group was 892 +/- 543 milliliters and for the PTFE group, 842 +/- 403 milliliters (p = NS). Patients in the Dacron group received intraoperatively 2.2 +/- 1.6 (units +/- S.D.) milliliters with a range of zero to 4 units of homologous liquid preserved erythrocytes and patients in the PTFE group received 1.2 +/- 1.2 milliliters with a range of zero to 3 units of homologous liquid preserved erythrocytes (p = NS). The mean total loss of erythrocytes (+/- S.D.) was 1,055 +/- 649 milliliters for the Dacron group and 978 +/- 503 milliliters for the PTFE group (p = NS). Despite inherent differences in graft material, there were no significant differences in intraoperative or post-operative loss of erythrocytes or in the number of homologous units of liquid preserved erythrocytes transfused with a p value of less than 0.05 considered significant.

Recovery of red blood cell mass following orthognathic surgery.

The rates of recovery of red blood cell mass after surgery were compared in three groups of orthognathic surgery patients: 12 patients not receiving transfusions during Le Fort I osteotomy, 10 patients receiving transfusions during Le Fort I osteotomy, and 25 patients undergoing bilateral sagittal split osteotomy. Patients in these groups lost a mean of 421, 455, and 191 mL of blood, respectively, during surgery. Both groups of patients undergoing Le Fort I osteotomy donated a unit of blood prior to surgery, while the patients undergoing bilateral sagittal split osteotomy did not. A control group consisted of 77 presurgical orthognathic patients who donated a unit of blood (450 mL) a mean of 11 days prior to surgery. Among the control group, 38 had recovered red blood cell mass prior to surgery. The patients undergoing Le Fort I osteotomy recovered red blood cell mass after surgery more quickly than did the patients undergoing bilateral sagittal split osteotomy, despite having lost twice as much blood. This unexpected finding can be attributed to the need to lose sufficient red blood cell mass to stimulate the erythropoietic system. The patients undergoing Le Fort I osteotomy experienced two challenges to the system, predonation of blood and greater blood loss during surgery, resulting in a great enough decrease in red blood cell mass to stimulate the release of erythropoietin. Patients undergoing bilateral sagittal split osteotomy did not exceed this threshold. None of the differences among the groups were statistically significant, possibly because of the few number of subjects.

Changes in hemoglobin and hematocrit levels following orthognathic surgery of the mandible.

Red blood cell mass has been shown to recover rapidly with great decreases, as occurs in orthognathic surgery. However, blood loss during bilateral sagittal split osteotomy is considered minimal. The purpose of this study was to verify changes in hemoglobin (Hb) and hematocrit (Ht) levels following orthognathic surgery of the mandible. Data from 12 patients with a mean age of 27.5 years, from both genders, that were submitted to bilateral sagittal split osteotomy in the prognathism reduction were evaluated. No patient received blood transfusion during the procedure nor postoperatively. Procedures were performed by the same surgeon, with the use of reciprocating and sagittal saws. Levels of Hb and Ht were obtained in preoperative, one day postsurgery, and one, three and six weeks postoperatively. Statistical analyses were used to verify the difference in each group, and between periods. Patients lost a mean decrease in Hb of 2.0 g/dl, and in Ht of 5.2%. There was significant difference as to the levels of Hb (p=0.002) and Ht (p=0.010). These levels presented significant changes from immediate postoperative to six weeks for Hb, and to three weeks for Ht. It was concluded that a slow recovery of hemoglobin and hematocrit levels occurred following orthognathic surgery of the mandible.

Swelling-stimulated passive potassium transport in camel erythrocytes: inhibitory effects of furosemide and sodium fluoride.

The inhibitory effects of furosemide, sodium fluoride, and age on volume-dependent, ouabain-resistant K+ influx were investigated in camel red blood cells. Swelling of young camel erythrocytes hypotonically stimulates ouabain-resistant potassium influx, a response that was lacking in old camel erythrocytes. The swelling-stimulated influx was partially inhibited by 1 mM furosemide and by 10 and 20 mM sodium fluoride. The inhibitory effect of furosemide was significantly increased if rubidium was added to the flux media. There was a significant correlation between potassium influx in normo- and hypotonic media which might indicate that the anion-dependent transport system operates, to some extent, to regulate cell volume.

A potent new dipeptide inhibitor of cell sickling and haemoglobin S gelation.

A dipeptide L-lysine-L-phenylalanine is shown to inhibit both cell sickling and the gelation of solutions of sickle-cell haemoglobin. The effect on deoxyhaemoglobin solutions and gels was followed by centrifugation; a progressive inhibition of gelation was observed up to 30 mM Lys-Phe. The haemoglobin concentration at the plateau (26 g/dl) suggests that an effect might be seen in vivo if suitable concentrations of Lys-Phe (about 20 mM) can be maintained in the blood stream. Additional studies of lag time before onset of gelation support this. Oxygen dissociation curves of sickle cells showed an effect of Lys-Phe only after incubation for 3 h before measurement, the P50 decreasing from 51 mmHg (6.8 MPa) to 41 mmHg (5.5 MPa) for cells depleted of 2,3-bisphosphoglycerate. The effect of Lys-Phe on intact sickle cells was more rapid. A marked increase in the number of unsickled cells in the presence of Lys-Phe was observed after only 15 min incubation. This result, together with measurements of uptake both into the cell and onto the cell membrane shows that the compound produces a membrane-mediated antisickling effect in addition to the effect on haemoglobin in solution within the cell. The membrane effect is not due to a change in cell volume. The properties of this dipeptide may be of value in the treatment of impending and early sickle crisis.