Microglial cells of the rat brain in postnatal period (comparative immunocytochemical analysis)

In the course of the brain’s development, distribution of microglial cells was studied in rats using immunocytochemical detection. To identify the microglial cells, antibodies to lipocortin 1 (LC1) and phosphotyrosine (PT) were used. On postnatal day 1, LC1-positive microglial cells of an ameboid shape were distributed mainly in the subventricular zone; their mean density was 31±8 cells/mm² (counted across the total area of frontal sections). On postnatal day 7, microglial cells of an intermediate type were located throughout the whole brain; their density was 54±15 cells/mm². On the 15th day, LC1-positive cells were of a ramified shape, and their density reached 104±20 cells/mm² (the microglial cell density in the mature normal brain was 103±3 cells/mm²). On postnatal day 7, PT-positive cells were similar in their morphology to LC1-positive cells of an intermediate type, while their mean density was 32 cells/mm². In the mature brain, the density of PT-positive microglia was 53±5 cells/mm²; the shape of the cells in the white and gray matter of the brain was, on the whole, similar to that of LC1-positive microglia. Therefore, LC1 is a specific marker for different types of microglial cells in the developing brain. Our data about 3D distribution and morphological peculiarities of microglial cells at different stages of postnatal development are consistent with the hypothesis on the neuroectodermal origin of microglia.     

Gender comparison of contractile performance and beta-adrenergic response in isolated rat cardiac trabeculae

It is known that gender can affect susceptibility to development of various cardiomyopathies. However, it is unclear whether basic mechanical contractile function of the myocardium differs between genders, whether they respond differently to stressors, or both. To test for a possible gender factor, contractile parameters of healthy, isolated myocardium were investigated under near physiological conditions. Right ventricular ultra-thin trabeculae from young adult LBN-f1 rats were electrically stimulated to isometrically contract at 37°C. No differences were found in developed force or kinetic parameters. In each muscle, the force-frequency relationship was measured at 4, 6, and 8 Hz, encompassing most of the in vivo range. Again, no differences were observed in force-frequency behavior; developed force rose from 21.6 ± 4.0 at 4 Hz to 30.3 ± 5.8 mN/mm² at 8 Hz in females and from 23.4 ± 3.4 to 29.8 ± 3.4 mN/mm² in males. The response to β-adrenergic stimulation was similar; at 1 μM isoproterenol, developed force increased to 34.5 ± 6.2 mN/mm² in females and 32.3 ± 3.2 mN/mm² in males (female vs. male, not significant). We conclude that basic mechanical performance of healthy isolated myocardium under physiological conditions is not different between males and females, and a different response to stress must underlie gender-based differences in cardiac performance.     

Structural diversity of nuptial pads in Phyllomedusinae (Amphibia: Anura: Hylidae)

We studied the morphological variation of the nuptial pads using light microscopy and scanning electron microscopy (SEM) in 26 species of phyllomedusines (Anura: Hylidae), representing the five currently recognized genera. All phyllomedusines have single nuptial pads with dark colored epidermal projections (EPs). Spine‐shaped EPs occur in Cruziohyla calcarifer, Phrynomedusa appendiculata and in one species of Phasmahyla. The other species have roundish EPs. The density of the EPs on the pad is variable. Species in the Phyllomedusa hypochondrialis Group have EPs with a density that varies between 764 ± 58/mm² and 923 ± 160/mm². In all other studied species (including the Phyllomedusa burmeisteri and Phyllomedusa perinesos groups, Phyllomedusa camba, Phyllomedusa boliviana, Phyllomedusa sauvagii, Phyllomedusa bicolor, and Phyllomedusa tomopterna) the density of EPs varies between 108 ± 20/mm² and 552 ± 97/mm². Pores were observed with SEM in C. calcarifer, Agalychnis lemur, Agalychnis moreletii, but its presence is confirmed through histological sections on several other species. Its visibility using SEM seems to be related with the level of separation between adjacent EPs. The pores in the four studied species of Agalychnis are shown with SEM and histological sections to have a characteristic epidermal rim, that is absent in the otherphyllomedusines. Unlike most previous reports on breeding glands, those of phyllomedusines are alcian blue positive, indicating the presence of acidic mucosubstances on its secretions. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc.     

High‐Performance Relaxor Ferroelectric Materials for Energy Storage Applications

Relaxor ferroelectrics usually possess low remnant polarizations and slim hystereses, which can provide high saturated polarizations and superior energy conversion efficiencies, thus receiving increasing interest as energy storage materials with high discharge energy densities and fast discharge ability. In this study, a relaxor ferroelectric multilayer energy storage ceramic capacitor (MLESCC) based on 0.87BaTiO₃‐0.13Bi(Zn_2/3(Nb_0.85Ta_0.15)_1/3)O₃ (BT‐BZNT) with inexpensive Ag/Pd inner electrodes is prepared by the tape casting method. The MLESCC with two dielectric layers (layer thicknesses of 5 µm) sintered by a two‐step sintering method exhibits excellent energy storage properties with a record‐high discharge energy density of 10.12 J cm^?3, a high energy efficiency of 89.4% achieved at an electric field of 104.7 MV m^?1, a high temperature stability of the energy storage density (with minimal variation of <±5%), and energy efficiency (>90%) over a range of ?75 to 150 °C at 40 MV m^?1. These results suggest that the BT‐BZNT relaxor ferroelectric ceramic material can provide realistic solutions for high‐power energy storage capacitors.     

Cuttlebone as a Marine-Derived Material for Preparing Bone Grafts

The use of synthetic materials for biomedical applications still presents issues owing to the potential for unfavourable safety characteristics. Currently, there is increasing interest in using natural, marine-derived raw materials for bone tissue engineering. In our study, the endoskeleton of the mollusc Sepia, i.e. cuttlebone (CB), was used with regenerated cellulose (RC) to prepare three-dimensional composite bone grafts. CB microparticles were mechanically immobilised within a cellulose gel, resulting in a macroporous structure upon lyophilisation. The interconnected porous structure of the regenerated cellulose/cuttlebone (RC/CB) composite was evaluated by micro-computed tomography. The porosity of the composite was 80%, and the pore size predominantly ranged from 200 to 500 μm. The addition of CB microparticles increased the specific scaffold surface by almost threefold and was found to be approximately 40 mm^?1. The modulus of elasticity and compressive strength of the RC/CB composite were 4.0?±?0.6 and 22.0?±?0.9 MPa, respectively. The biocompatibility of the prepared RC/CB composite with rat hepatocytes and extensor digitorum longus muscle tissue was evaluated. The obtained data demonstrated that both the composite and cellulose matrix samples were non-cytotoxic and had no damaging effects. These results indicate that this RC/CB composite is a novel material suitable for bone tissue-engineering applications.     

Effects of successivein vitro andIn vivo passages on the virulence of the entomopathogenic fungus,Nomuraea rileyi

There was no significant decrease or increase in the activity of conidia ofNomuraea rileyi after 12_{in vivo} serial passages in larvae ofTrichoplusia ni (Hübner) or after 12in vitro serial passages on a semi-synthetic medium. The LC-50 at the start of the serial passage was 16.8±4.0 conidia/mm²; after 12 serial passages the LC-50 for thein vitro- andin vivo-produced conidia was 16.2±2.5 conidia/mm² and 12.0±1.9 conidia/mm², respectively.

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Résumé Il n'y a pas d'augmentation ou de réduction significative dans l'activité des conidies deNomuraea rileyi après 12 passages en sériein vivo chez les larves deTrichoplusia ni (Hübner) ou après 12 passages en sériein vitro sur un milieu semi-synthétique. La concentration léthale 50 au début de ces passages était de 16,8±4,0 conidies/mm², après 12 passages la CL 50 des conidies produitesin vitro etin vivo fut respectivement de 16,2±2,5 conidies/mm² et de 12,0±1,9 conidies/mm².

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Mention of a proprietary product in this paper does not constitute a recommendation for use by the USDA.     

Microstructure,morphology and physicochemical properties of nanocomposites containing hydroxyapatite/vivianite/graphene oxide for biomedical applications

Designing a nanocomposite that accumulates biocompatibility and antimicrobial behaviour is an essential requirement for biomedical applications. Hydroxyapatite (HAP), graphene oxide, and vivianite in one ternary nanocomposite with three phases and shapes led to an increase in cell viability to 97.6% ± 4 for the osteoblast cells in vitro. The obtained nanocomposites were investigated for their structural features using X-ray diffraction, while the microstructure features were analyzed using a scanning electron microscope (SEM) and a transmission electron microscope. The analysis showed a decrease in the crystal size to 13 nm, while the HAP grains reached 30 nm. The elongated shape of vivianite reached 200 nm on SEM micrographs. The monoclinic and hexagonal crystal systems of HAP and vivianite were presented in the ternary nanocomposite. The maximum roughness peak height reached 236.1 nm for the ternary nanocomposite from 203.3 nm, while the maximum height of the roughness parameter reached 440.7 nm for the di-nanocomposite of HAP/graphene oxide from 419.7 nm. The corrosion current density reached 0.004 μA/cm². The ferrous (Fe²⁺) and calcium (Ca²⁺) ions released were measured and confirmed. Therefore, the morphology of the nanocomposites affected bacterial activity. This was estimated as an inhibition zone and reached 14.5 ± 0.9 and 13.4 ± 1.1 mm for Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) after 24 h. The increase in viability and the antibacterial activity refer to the compatibility of the nanocomposite in different medical applications.     

In situ characterization of mast cells in the frog Rana esculenta

The number, distribution, and ultrastructural characteristics of mast cells were assessed in the tongue, heart, and kidney of the frog Rana esculenta. The density of tongue mast cells (253±45 mast cells/mm²) was significantly higher than that of the heart (5.3±0.4/mm²) and kidney (15.3±1.4 /mm²). A striking feature of this study was the remarkable association of frog mast cells to nerves. The ultrastructural study of the mast cell/nerve association demonstrated that mast cells were closely apposed to or even embedded in nerves. Mast cells were also physically associated with melanocytes even in the heart. Mast cells were Alcian blue⁺/safranin⁺ in the tongue and in the peritoneum, whereas in the heart and in the kidney they were Alcian blue^–/safranin⁺. The mast cells in the lamina propria of the gastrointestinal tract were Alcian blue⁺/safranin^–. The cytoplasm of frog mast cells was packed with numerous heterogeneous, membrane-bound granules. The ultrastructure of these cytoplasmic granules was unique, being totally unlike any other previously described granules in other animal species as well as in man. The histamine content/frog mast cell (≈0.1 pg/cell) was approximately 30 times lower than that of human mast cells isolated from different tissues (≈3 pg/cell). A monoclonal anti-histamine antibody was used to confirm the ultrastructural localization of histamine in secretory granules in frog mast cells.     

Genetic analysis of barrel field size in the first somatosensory area (SI) in inbred and recombinant inbred strains of mice

We measured the combined area of posterior medial barrel subfield (PMBSF) and anterior lateral barrel subfield (ALBSF) areas in four common inbred strains (C3H/HeJ, A?/J, C57BL?/6J, DBA/2J), B6D2F1, and ten recombinant inbred (RI) strains generated from C57BL/6J and DBA/2J progenitors (BXD) as an initial attempt to examine the genetic influences underlying natural variation in barrel field size in adult mice. These two subfields are associated with the representation of the whisker pad and sinus hairs on the contralateral face. Using cytochrome oxidase labeling to visualize the barrel field, we measured the size of the combined subfields in each mouse strain. We also measured body weight and brain weight in each strain. We report that DBA/2J mice have a larger combined PMBSF/ALBSF area (6.15?±?0.10?mm²,?n?=?7) than C57BL?/6J (5.48?±?0.13?mm²,?n?=?10), C3H/HeJ (5.37?±?0.16?mm²,?n?=?10), and A/J mice (5.04?±?0.09?mm²,?n?=?15), despite the fact that DBA/2J mice have smaller average brain and body sizes. This finding may reflect dissociation between systems that control brain size with those that regulate barrel field area. In addition, BXD strains (average n?=?4) and parental strains showed considerable and continuous variation in PMBSF/ALBSF area, suggesting that this trait is polygenic. Furthermore, brain, body, and cortex weights have heritable differences between inbred strains and among BXD strains. PMBSF/ALBSF pattern appears similar among inbred and BXD strains, suggesting that somatosensory patterning reflects a common plan of organization. This data is an important first step in the quantitative genetic analysis of the parcellation of neocortex into diverse cytoarchitectonic zones that vary widely within and between species, and in identifying the genetic factors underlying barrel field size using quantitative trait locus (QTL) analyses.     

兔前交叉韧带损伤后膝关节本体感觉、残端血运及膝关节腔尿激酶型纤溶酶原激活物变化的实验研究

摘要 目的：研究兔前交叉韧带（ACL）损伤后膝关节本体感觉、残端血运及膝关节腔尿激酶型纤溶酶原激活物（u-PA）的变化情况。方法：选取80只新西兰兔进行研究,将其以随机数字表法分作模型组及空白对照组各40只。模型组建立单侧ACL损伤模型,空白对照组仅切开关节。比较两组术前及术后2周、4周、8周时膝关节本体感觉、残端血运及膝关节腔u-PA水平的差异。结果：模型组新西兰兔术后2周、4周、8周时的体感诱发电位（SEP）、肌电图（EMG）潜伏期均高于空白对照组,而SEP、EMG波幅均低于空白对照组（P＜0.05）。模型组新西兰兔术后2周、4周、8周时的残端组织微血管密度分别为（2.04±0.24）n/mm²、（2.75±0.61）n/mm²、（1.60±0.33）n/mm²,均高于空白对照组的（1.34±0.24）n/mm²、（1.34±0.25）n/mm²、（1.35±0.26）n/mm²,差异均有统计学意义（P＜0.05）。模型组新西兰兔术后2周、4周、8周时的膝关节液u-PA水平分别为（173.97±14.29）pg/mL、（188.37±15.82）pg/mL、（171.38±14.76）pg/mL,均高于空白对照组的（158.02±10.18）pg/mL、（157.68±10.20）pg/mL、（157.37±10.07）pg/mL,差异均有统计学意义（P＜0.05）。结论：ACL损伤后会在不同程度上影响膝关节本体感觉、残端血运及膝关节腔u-PA含量,值得临床进一步研究。     

A losing battle: weight regain does not restore weight loss-induced bone loss in postmenopausal women

Previously, we reported significant bone mineral density (BMD) loss in postmenopausal women after modest weight loss. It remains unclear whether the magnitude of BMD change in response to weight loss is appropriate (i.e., proportional to weight loss) and whether BMD is recovered with weight regain. We now report changes in BMD after a 1‐year follow‐up. Subjects (n = 23) in this secondary analysis were postmenopausal women randomized to placebo as part of a larger trial. They completed a 6‐month exercise‐based weight loss program and returned for follow‐up at 18 months. Dual‐energy X‐ray absorptiometry (DXA) was performed at baseline, 6, and 18 months. At baseline, subjects were aged 56.8 ± 5.4 years (mean ± s.d.), 10.0 ± 9.2 years postmenopausal, and BMI was 29.6 ± 4.0 kg/m². They lost 3.9 ± 3.5 kg during the weight loss intervention. During follow‐up, they regained 2.9 ± 3.9 kg. Six months of weight loss resulted in a significant decrease in lumbar spine (LS) (?1.7 ± 3.5%; P = 0.002) and hip (?0.04 ± 3.5%; P = 0.03) BMD that was accompanied by an increase in a biomarker of bone resorption (serum C‐terminal telopeptide of type I collagen, CTX: 34 ± 54%; P = 0.08). However, weight regain was not associated with LS (0.05 ± 3.8%; P = 0.15) or hip (?0.6 ± 3.0%; P = 0.81) bone regain or decreased bone resorption (CTX: ?3 ± 37%; P = 0.73). The findings suggest that BMD lost during weight reduction may not be fully recovered with weight regain in hormone‐deficient, postmenopausal women. Future studies are needed to identify effective strategies to prevent bone loss during periods of weight loss.     

Mid-term function and remodeling potential of tissue engineered tricuspid valve: Histology and biomechanics

ObjectiveTricuspid valve reconstruction using a small intestinal submucosal porcine extracellular matrix (ECM) tube graft is hypothesized to be durable for six months and show signs of recellularization and growth potential. The purpose was to histologically and biomechanically test ECM valves before and after six months of implantation in pigs for comparison with native valves.MethodsTen 60 kg pigs were included, which survived tricuspid valve tube graft insertion. Anterior and septal tricuspid leaflets were explanted from all animals surviving more than one month and examined histologically (n = 9). Endothelialization, collagen content, mineralization, neovascularization, burst strength and tensile strength were determined for native valves (n = 5), ECM before implantation (n = 5), and ECM after six months (n = 5).ResultsCollagen density was significantly larger in ECM at implantation (baseline) compared to native leaflet tissue (0.3 ± 0.02 mg/mm³ vs. 0.1 ± 0.03 mg/mm³, p < .0001), but collagen density decreased and reached native leaflet collagen content, six months after ECM implantation (native vs. ECM valve at six months: 0.1 ± 0.03 mg/mm³ vs. 0.2 ± 0.05 mg/mm³, p = .8).Histologically, ECM valves showed endothelialization, host cell infiltration and structural collagen organization together with elastin generation after six months, indicating tissue remodeling and -engineering together with gradual development of a close-to-native leaflet structure without foreign body response.ConclusionsECM tricuspid tube grafts were stronger than native leaflet tissue. Histologically, the acellular ECM tube grafts showed evidence of constructive tissue remodeling with endothelialization and connective tissue organization. These findings support the concept of tissue engineering and recellularization, which are prerequisites for growth.     

Short-term parenteral application of α-tocopherol leads to increased concentration in plasma and tissues of the rat

Numerous studies suggest that supplemental vitamin E prior to or during vast surgeries might diminish or even prevent ischemia/reperfusion-induced injuries. In the present placebo-controlled study male Sprague-Dawley rats were supplemented parenterally or orally with α-tocopherol for three consecutive days. The applied amount of α-tocopherol was 2.3 μmol per day for oral and 1.2 μmol per day for parenteral supplementation. The enrichment of vitamin E concentrations in plasma and tissue samples (aortic endothelium, liver, and lung) was determined by HPLC. The vitamin E level was elevated following intravenous supplementation in plasma (21.4±1.9 μmol/L vs. 10.2±1.7 μmol/L in parenteral control group), in aortic endothelium (1.1±0.2 pmol/mm² vs. 0.5±0.1 pmol/mm²) and in liver and lung (41.3±7.5 pmol/mg vs. 22.9±6.5 pmol/mg and 75.6±13.6 pmol/mg vs. 51.7±5.9 pmol/mg, respectively). Oral supplementation for three days also led to an increased level in liver (38.2±7.7 pmol/mg vs. 22.9±6.6 pmol/mg in oral control group) and in lung (67.8±5.7 pmol/mg vs. 51.7±9.3 pmol/mg) but not in aortic endothelium or plasma (0.8±0.3 pmol/mm² vs. 0.6±0.3 pmol/mm² and 12.0±2.2 μmol/L vs. 10.7±2.6 μol/L.)     

Estimating Activity‐related Energy Expenditure Under Sedentary Conditions Using a Tri‐axial Seismic Accelerometer

Activity‐related energy expenditure (AEE) is difficult to quantify, especially under sedentary conditions. Here, a model was developed using the detected type of physical activity (PA) and movement intensity (MI), based on a tri‐axial seismic accelerometer (DynaPort MiniMod; McRoberts B.V., The Hague, the Netherlands), with energy expenditure for PA as a reference. The relation between AEE (J/min/kg), MI, and the type of PA was determined for standardized PAs as performed in a laboratory including: lying, sitting, standing, and walking. AEE (J/min/kg) was calculated from total energy expenditure (TEE) and sleeping metabolic rate (SMR) as assessed with indirect calorimetry ((TEE × 0.9) ‐ SMR). Subsequently, the model was validated over 23‐h intervals in a respiration chamber. Subjects were 15 healthy women (age: 22 ± 2 years; BMI: 24.0 ± 4.0 kg/m²). Predicted AEE in the chamber was significantly related to measured AEE both within (r² = 0.81 ± 0.06, P < 0.00001) and between (r² = 0.70, P < 0.001) subjects. The explained variation in AEE by the model was higher than the explained variation by MI alone. This shows that a tri‐axial seismic accelerometer is a valid tool for estimating AEE under sedentary conditions.     

Lethal and sub‐lethal impacts of pulsed laser irradiations on the larvae of the fouling barnacle Balanus amphitrite

Laboratory experiments were conducted to study the impact of laser irradiation on the larvae of the fouling barnacle Balanus amphitrite. Research pertaining to fouling invertebrate larvae‐laser interaction is sparse and, hence, data on this aspect were thought significant in order to consider pulsed low power laser irradiations as a possible future antifouling tool. Lethal and sub‐lethal impacts of four very low laser fluences, viz. 0.013, 0.025, 0.05 and 0.1 J cm^‐2 for three different durations, viz. 2, 10 and 30 s were investigated. Three growth stages of barnacle larvae, viz. nauplii stage II, nauplii stage IV and cyprids were exposed to the mentioned laser fluences for different durations. While lethal impact was assessed immediately after and 1 d after irradiation, sub‐lethal impacts were studied by monitoring the success rate of the irradiated nauplii in reaching the cyprid stage. In addition, the swimming speed of VI^th stage nauplii after irradiation was studied. In the case of cyprids, in addition to the mortality measurement immediately after and 1 d after irradiation, the settlement rate was investigated. In all the above experiments, non‐irradiated larvae served as controls. The results showed an increase in mortality with increasing laser fluence and duration of irradiation. Irradiation for 2 s resulted in significant mortality in nauplii, while it was less in the case of cyprids. In II^nd stage nauplii, the mortality immediately after irradiation for 2 s varied from 14.8±2.12 to 97.1±4.1% for laser fluences of 0.013 and 0.1 J cm^‐2, respectively. However, in cyprids, the mortality immediately after irradiation for 2 s varied from 12.2±3 to 13.4±1.2% for fluences of 0.013 and 0.1 J cm^‐2, respectively. The mortality in IV^th stage nauplii was less than that for II^nd stage nauplii but more than that for cyprids. There was a significant increase in mortality with time after irradiation. The formation of cyprids from the irradiated larvae was significantly less than that observed for non‐irradiated larvae. Also, the irradiated larvae showed a significantly slower swimming speed compared to the control samples. The settlement rate in cyprids was reduced significantly by the laser irradiation. This was true even for the lowest fluence and shortest period of irradiation tested. Thus, the results of the experiment showed that even a low power pulsed laser irradiation of 0.013 J cm^‐2 for 2 s can cause significant damage to fouling barnacle larvae.     

Kinetics of picosecond bacteriochlorophyll luminescence in vivo as a function of the reaction center state

Bacteriochlorophyll (BChl) luminescence lifetimes (τ) were measured in purple bacteria Rhodospirillum rubrum and Rhodopseudomonas sphaeroides at low-excitation pulse energy with the use of a picosecond luminescence spectrochronograph of high sensitivity and high time-resolution. Average high-frequency excitation light density was changed from about 1 · 10¹³ photons · cm⁻² · s⁻¹ up to 1 · 10¹⁷ photons · cm⁻². s⁻¹. Maximal energy density in a single pulse was in the range 10⁻¹⁴–10⁻¹⁰ J/cm², which completely rules out nonlinear exciton interactions. In this range τ increased as a function of excitation light density from about 60 ps to 210 ps. Luminescence yield (ø) for the bacteria investigated measured under continuous or picosecond excitation changed in a similar manner as τ. The luminescence increase was shown to accompany the conversion of the reaction centers to the closed, photooxidized state. Luminescence decay of R. rubrum and Rps. sphaeroides chromatophores without any chemical additions was well approximated by a single exponential component both at low and at saturating intensities of exciting light. The time necessary for the primary charge separation to occur was shown to be 60 ± 10 ps. The pairwise jump-time of excitation-energy transfer, as well as excitation-diffusion characteristics were estimated from these data. On the basis of life-time measurements in the state of active photosynthesis, the quantum yield of the primary charge separation in the reaction centers was estimated to be equal to 0.95 ± 0.02. In intact cells as well as in chromatophores in the presence of reducing agents, a nanosecond component of emission decay was also observed. The relative amplitude of this component, being several percent of the picosecond one at low-excitation intensity levels, increased (2–3)-times with excitation density. Its life-time was estimated to be 3 ± 1 ns. The nanosecond component appeared only under conditions when a part of the reaction centers were converted to the closed state PQ⁻.     

In vitro biomechanical study of female geriatric cervical vertebral bodies

Compressive strength tests were conducted on fresh human geriatric female cervical vertebral bodies. Nineteen specimens were compressed to 50% of their initial height using an electrohydraulic testing device. The mechanical force-deflection response was sigmoidal with continuously changing resistance. The mean cross-sectional area and bone mineral content (BMC) of the vertebral bodies progressively increased from C₃ (area: 333.8 mm², BMC: 1.56 g) to C₆ (area: 499.7 mm², BMC: 2.18 g). The maximum compressive force increased from 1060 N at C₃ to 1787 N at C₆. The stiffness and the energy absorbed at failure also increased from C₃ to C₆ (stiffness: 279.95 to 556.41 N mm⁻¹, energy: 2.45 to 4.16 J). These parameters demonstrated a decreasing tendency from C₆ to C₇. However, the mean failure deformations did not exhibit a significant trend with respect to spinal level from C₃ to C₇. The relatively higher biomechanical parameters at the sixth vertebral level compared with its caudad and cephalad counterparts may be due to the fact the transition of the cervical lordosis to thoracic kyphosis begins at this level. Furthermore, the change in the anatomy of the uncinate processes in the cervical column around this region may also be a contributing factor.     

Distribution of microglia and astrocytes in different regions of the normal adult rat brain

The distribution of glial cells (microglia and astrocytes) in different regions of normal adult rat brain was studied using immunohistochemical techniques and computer analysis. Antibodies against lipocortin 1 (LC1) and phosphotyrosine (PT), as well as an isolectin, GSA B4 (GSA), were used for identification of microglial units, while antibodies against protein S100β allowed us to identify astrocytes. If LC1 was used as a marker, more microglial cells were detected than with the use of PT or GSA. The highest density of LC1-positive microglial cells (on average, 130±5 cells/mm² of the brain section area) was found in the neostriatum, while the lowest density (51±4 cells/mm²) was observed in the medulla oblongata. In general, the density of an LC1-positive microglial population was higher in the forebrain and lower in the midbrain, and the smallest number of these cells was detected in the brainstem and cerebellum. The number of astrocytes was, on average, 2–3 times as large as the number of microglial cells. High density of astrocytes, was found in the hypothalamus and hippocampus (more than 260 cells/mm²); they were more, numerous in the white matter than in the gray matter. Lower densities of this type cells were observed in the cerebral cortex, neostriatum, midbrain, medulla oblongata, and cerebellum (less than 200 cells/mm²).     

Endothelial quality of pre-cut posterior corneal lamellae for Descemet membrane endothelial keratoplasty with a stromal rim (DMEK-S): two-year outcome of manual preparation in an ocular tissue bank

To assess the quantitative and qualitative parameters of pre-cut posterior corneal lamellae for Descemet membrane endothelial keratoplasty with a stromal rim (DMEK-S) prepared manually in the Ocular Tissue Bank Prague. All 65 successfully prepared pre-cut posterior corneal lamellae provided for grafting during a 2-year period were analyzed retrospectively. The lamellae, consisting of a central zone of endothelium-Descemet membrane surrounded by a supporting peripheral stromal rim, were prepared manually from corneoscleral buttons having an endothelial cell density higher than 2,500 cells/mm². The live endothelial cell density, the percentage of dead cells, the hexagonality and the coefficient of variation were assessed before and immediately after preparation as well as after 2 days of organ culture storage at 31 °C. Altogether, the endothelium of 57 lamellae was assessed. Immediately after preparation, the mean live endothelial cell density was 2,835 cells/mm² and, on average, 1.8 % of dead cells were found. After 2 days of storage, the cell density decreased significantly to 2,757 cells/mm² and the percentage of dead cells to 1.0 %. There was a significant change in the mean hexagonality and the coefficient of variation after lamellar preparation and subsequent storage. The amount of tissue wasted during the preparation was 23 %. The endothelial cell density of posterior corneal lamellae sent for DMEK-S was higher than 2,700 cells/mm² in average with a low percentage of dead cells; 65 pre-cut tissues were used for grafting during a 2-year period.