Microdensitometric Analysis Research Articles

Neuropathic or systemic chronic intoxication with Karwinskia humboldtiana (Buckthorn) fruit? Histopathological effect in myocardial and skeletal muscle

In accidental intoxicated animals and humans, Karwinskia humboldtiana (Kh) causes lesions in the central and peripheral nervous system and organs like the kidney, liver, and lung. The objective was to evaluate the histology of myocardium and skeletal muscle after experimental chronic intoxication with mature fruit of Kh in Wistar rat. Twenty-five rats were used and divided into five groups (n = 5): four intoxicated and one control. Kh fruit was ground, dried, sieved, and administered by an orogastric tube. Intoxicated rats received 3.5 g/kg body weight fractionated in 5 doses. Control rats received only water. Animals were euthanized at 24, 48, 58, and 112 days, respectively. Samples of the myocardium and skeletal muscle were obtained and processed for light microscopy evaluation. Morphological analyses were performed, including a microdensitometric analysis. Results showed areas of necrosis in the muscle fibers, fibers with vacuolated cytoplasm, and disorganization of myofilaments, as well as staining variations in both myocardium and skeletal muscle time-depending. Zones with loss of continuity of the external lamina were identified with PAS with the diastase histochemical method. Immunolabeling with specific antibodies demonstrated diminution of actin and desmin myofilaments. The microdensitometric analysis showed a statistically significant difference between the intoxicated vs control group. These findings demonstrate that chronic intoxication of Kh fruit also causes damage in myocardial and skeletal muscle, these alterations will be useful to understand that the toxic effects of Kh fruit in accidently intoxicated humans are systemic, and not only over the nervous system.

Physical, chemical and mechanical properties of Pinus sylvestris wood at five sites in Portugal

Abstract: The reduction of resinous species in Portuguese forest areas has caused constraints to wood industry supplies. Portugal represents the extreme southwest of Scots pine (Pinus sylvestris) natural distribution and large gaps exist in the knowledge of its wood-quality characteristics. Understanding the relationship between these traits is important for recognizing which combination of wood properties is the most suitable for specific uses. To address these questions, we assessed wood-quality traits, namely, wood-density components (microdensitometric analysis), chemical composition (NIR spectrometry) and mechanical properties (bending tests) of wood samples collected at five representative forest sites in Portugal. Our results showed that Portuguese Pinus sylvestris has good radial growth and denser wood, higher extractive content and higher stiffness and strength than northern European provenances. The lignin content was within the range attributed to softwoods. Among the Portuguese stands, trees growing at lower-altitude sites exhibited denser wood and higher mechanical properties, while trees from high-elevations showed higher amounts of lignin. Ring density was more strongly correlated with earlywood than latewood density. A negative, non-significant correlation was found between ring density and width, supporting the assumption that the higher radial growth (ring width) does not negatively affect wood quality (density). In general, chemical properties had a weak relationship with physical and mechanical properties (MOE and MOR). Both mechanical traits were positively correlated with density and growth components, supporting the assumption that trees with high radial growth do not exhibit poorer mechanical performances.

Growth ring analysis of Euxylophora paraensis through x-ray microdensitometry

ABSTRACT: Euxylophora paraensis Huber - Rutaceae, popularly known as yellow heart, is a species with a low-density population that has suffered intense exploitation in recent decades. This has led to its inclusion in the IBAMA list of endangered species. This study aimed to evaluate the existence of growth rings and their annuity in five trees of Euxylophora paraensis through the x-ray microdensitometry technique, as well as to compare them using the classical method. Application feasibility of this technique can be deepened through future, broader dendroecological studies, which may better elucidate the understanding of the species population dynamics and promote more sustainable uses. Wood disks were collected from the base of five 29-year-old trees from a planting site in the Curua-Una Experimental Station, located in Prainha, Pará State, Brazil. Samples were polished and sanded for the x-ray microdensitometric analysis. Values of minimum, average, and maximum density (0.581, 0.84, and 1.077gcm-3, respectively) were obtained. There was a significant difference in density between the studied trees. Growth rings boundaries, through densitometric pith-bark profile are demarcated by fibrous zones, with density variation between the early wood and late wood. This study concludes that the growth rings of the studied species are distinct and annual, validating the technique used.

Morphological and histomorphometric evaluation of the ventral rectus sheath of the rectus abdominis muscle, fascia lata and pectoral fascia. The beginning of a morphological information bank of human fascias.

The aim of this study was to characterize and compare the morphological and histomorphometric characteristics of the pectoral fascia, fascia lata and ventral rectus sheath. Twenty cadaveric samples of these fascias were analyzed and stained with hematoxylin and eosin, orcein, Van Gieson, Masson's trichrome and Verhoeff¨s stain (1200 slides in total). Morphological evaluation, semiquantitative, morphometric and microdensitometric analysis of elastic fibers present in each of the tissues and a morphometrical analysis of tissue thickness were performed. The mean value of the pectoral fascia thickness was 612±68.13 μm; 84±246 μm for the fascia lata and 584±92 μm for the ventral rectus sheath. The area occupied by the elastic fibers in the pectoral fascia was 12.24±5.84%; 6,54±3.85% for the fascia lata and 11.11±5.26% for the ventral rectus sheath. There were no statistically significant differences when comparing the mean values between the pectoral fascia and the ventral rectus sheath (p=0.07). There were statistically significant differences when comparing the fascia lata to the pectoral fascia and the ventral rectus sheath (p≤0.001). This study reports other morphological characteristics not described in previous histological studies of the analyzed tissues. The results of the morphometric and densitometric analysis in this study reveal that the fascia lata has the fewest elastic fibers of all the tissues analyzed, and the pectoral fascia has the most. These results will be useful for the beginning of a morphological information bank of human fascias.

Oil palmin vitroregeneration: microdensitometric analysis during reproduction and development

An extensive microdensitometric analysis was performed in oil palm Elaeis guineensis Jacq. during the establishment of a new embryogenic culture, on new regenerated plants at different times, and f...

The novel cyclooxygenase-2 inhibitor GW637185X protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity

The possible neuroprotective role of a novel and highly selective cyclooxygenase-2 inhibitor GW637185X was studied in a model of acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced injury of nigrostriatal dopaminergic (DA) neurons in the mouse. Stereological and microdensitometrical analysis of nigral tyrosine hydroxylase-immunoreactive cell bodies and striatal tyrosine hydroxylase-immunoreactive terminals, respectively, showed that GW637185X exerted a full protection against MPTP-induced degeneration of the nigro-striatal pathway. In contrast to earlier studies, these findings demonstrate that acute inhibition of cyclooxygenase-2 can result in a full neuroprotective effect not only on nigral DA cell bodies, but also on striatal DA terminals in the mouse MPTP model.

Non-random distribution of high density chromatin detected at opposite ends of T-banded human metaphase chromosomes

Previous research using microdensitometric scanning and computer graphic image analysis showed that T-banded segments of human metaphase chromosomes usually exhibit an asymmetrical distribution of high density (HD) chromatin between sister chromatids. Here, we employed the same methods to analyze HD chromatin distribution at opposite ends of T-banded human lymphocyte chromosomes. This study revealed that in most chromosomes with an asymmetrical distribution of HD chromatin at both ends, the highest densities of each arm were located in opposite chromatids. The frequency of this configuration was 0.792 per chromosome, indicating that the highest chromatin densities of the terminal segments of T-banded human chromosomes were non-randomly distributed at opposite chromosome arms. The possible relationship of this observation to the mode of replication of the terminal chromosome region is briefly discussed.

Composition, distribution and supposed origin of mineral inclusions in sessile oak wood — consequences for microdensitometrical analysis

SEM and light-microscopical observations, supported by chemical microanalysis with an EDXA system, revealed that light-saturated pixels observed in X-ray negatives of sessile oak (Quercus petraea Liebl.) wood were caused by inorganic deposits present inside multiseriate ray and axial parenchyma cells. Calcium oxalate crystals, silica grains and amorphous granules with varied mineral compositions have been identified. The wood strips of three out of six sampled trees contained measurable amounts of mineral inclusions which were quantified using image analysis. Based on the variations of mineral content observed between trees and within and between annual rings of the same tree, some hypotheses were formulated concerning the factors involved in the formation of inorganic deposits in oak wood. Their occurrence varies depending on the mineral concerned and seems to be controlled largely by a tree effect. The time of formation appears to coincide with a shifting of the oak wood’s functions as a result of heartwood formation processes (inter-annual scale) or changes in leaf phenology and climate (intra-annual scale). In addition, the technical consequences of their presence as well as their effects on wood density measurements through microdensitometry are discussed.

Traumatic cataract and other adverse events with the implantable contact lens

To evaluate the surgical outcome and adverse events associated with implantation of the implantable contact lens (ICL) for the correction of high myopia or high hyperopia. Helsinki University Eye Hospital, Helsinki, Finland. In this nonrandomized prospective clinical trial, the ICL V4 was implanted in 26 eyes of 13 patients with normal enrollment criteria (younger than 45 years, a clinically clear crystalline lens) (Group 1) and the V2, V3, or V4 were implanted in 38 eyes of 22 patients with heterogeneous criteria (older than 45 years or opacities in the crystalline lens) (Group 2). The main outcome measures were lens opacity classification system (LOCS II) assessment and transparency analysis of the lens. In Group 1, microdensitometric assessment showed no decrease in crystalline lens transparency and no statistically significant increase in crystalline lens density at any follow-up examination (P>.05). The incidence of anterior subcapsular (AS) opacities was 7.7%, and no eye developed clinically significant cataract during the mean follow-up of 13.2 months +/- 12.3 (SD). No myopic eye lost a line of best corrected visual acuity (BCVA); 50.0% of hyperopic eyes lost 1 line of BCVA. Sixty-eight percent of myopic eyes and 75.0% of hyperopic eyes were within +/-1.0 diopter of the predicted refraction. In Group 2, the incidence of AS opacities was 47.4% and 10 eyes (26.3%) developed clinically significant cataract during the mean follow-up of 30.9 +/- 18.0 months. The visual and refractive results were similar to those in Group 1 at 1 month, but due to progressive AS opacities in the crystalline lens, 23.4% of eyes lost BCVA lines during the follow-up. Intraoperative complications were few and insignificant. Implantation of the ICL for correction of high myopia or high hyperopia appears to be a safe procedure with good visual and refractive results immediately. There was no decrease in the transparency or increase in the density of the crystalline lens with the latest generation of the ICL device in the normal group, indicating enhanced safety. Progressive lens opacification in the heterogeneous group shows there are risk factors for cataract formation; ie, type V3 ICL, preexisting lens opacities, and older patient age. Anterior subcapsular opacities are most likely the result of trauma.

Chromosome Banding and DNA Methylation Patterns, Chromatin Organisation and Nuclear DNA Content in Zingeria biebersteiniana

Chromosome structure and chromatin organisation of a two-chromosome model cereal Zingeria biebersteiniana (Claus) P. Smirnov were studied: nuclear DNA content was determined by microdensitometric analysis after Feulgen staining; Feulgen absorption at different thresholds of absorbance in interphase nuclei also provided evidence on the organisation of chromatin, allowing quantitative estimation of condensed chromatin within interphasic nucleus. The DNA methylation pattern of Z. biebersteiniana metaphase chromosomes was examined with a specific monoclonal antibody. 5-methyl-cytosine residues are present in several chromosome sites and differences may be present between corresponding regions of homologues. Chromosome banding pattern reveals large bands in the centromeric regions of each chromosome, showing constitutive heterochromatin; by fluorochromes staining pericentromeric blocks are evidenced. After the cold and 9-aminoacridine pre-treatments and after aceto-carmine and aceto-orceine staining, respectively, the metaphase chromosomes were analysed by image analysis system revealing a segmentation of the chromosome body that resembles Giemsa/Reverse banding in animal chromosomes.

Expression of pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) in sickle cell retina and choroid

Pigment epithelium-derived factor (PEDF) has been shown to be an inhibitor of angiogenesis as well as a multipotent neurotrophic factor in the mammalian eye. Changes in PEDF levels have been correlated with development of retinal neovascularization in oxygen-induced retinopathy. The purpose of this study was to determine the localization and relative level of PEDF in human retinas and choroids using immunohistochemistry and evaluate the changes in PEDF and vascular endothelial growth factor (VEGF) localization and their relation to the progression of proliferative sickle cell retinopathy. Cryopreserved tissues from eyes of normal subjects and subjects with non-proliferative or proliferative sickle cell retinopathy were used with streptavidin peroxidase immunohistochemistry. A rabbit polyclonal antibody was made against recombinant human PEDF. Binding of the antibody was blocked by preincubation of the antibody with excess human recombinant PEDF. Relative levels of immunoreactivity were scored with a seven-point grading system and by microdensitometric analysis. The most prominent sites of PEDF localization in the normal eye were the vitreous condensed at the internal limiting membrane and RPE–Bruch's membrane–choriocapillaris complex. PEDF was also prominent in choroidal stroma. There was limited immunoreactivity in some cells of the neural retinas, in blood vessels and in the interphotoreceptor matrix (IPM). There was no difference in ratio (1·47 vs. 1·44) of PEDF/VEGF or the relative levels of either growth factor in the retinal vasculatures of the control subjects and perfused area of non-proliferative sickle cell retinas. The ratio was increased in the non-perfused area of the non-proliferative sickle cell retinas (2·24). In eyes with proliferative sickle cell retinopathy, elevated PEDF and VEGF immunostaining was present in viable vessels of sea fan neovascular formations as well as feeder vessels of sea fans. The PEDF/VEGF ratio in sea fans was 1·0. Immunoreactivity for PEDF was prominent in retinal vessels in non-perfused regions and in atrophic sea fans, while VEGF immunoreactivity was weak or absent in these structures. In conclusion, PEDF and VEGF were both significantly elevated in viable sea fan formations in sickle cell disease ( p<0·05) but only PEDF was present in non-viable sea fans. The highest levels of PEDF in all eyes were associated with extracellular matrices (vitreous, choroidal stroma, IPM, and walls of blood vessels). PEDF might play an important role in inhibiting angiogenesis and inducing the regression of sea fans. Progression of angiogenesis may be dependent on the ratio of PEDF/VEGF.

Chronic nicotine treatment changes the axonal distribution of 68 kDa neurofilaments in the rat ventral tegmental area.

Region-specific decreases of neurofilament proteins (NF) were described in the ventral tegmental area (VTA) of rats treated chronically with morphine, cocaine or alcohol. In a previous study, we demonstrated that NF levels were also changed in the VTA after chronic treatment with nicotine. The aim of this study was to clarify the submicroscopic basis of decreased immunoreactivity for NF-68, NF-160 and NF-200, as determined by using NR4, BF10 and RT97 antibodies, respectively. Microdensitometric analysis of brain sections showed that immunoreactivity for all NF was reduced in the VTA of animals exposed chronically to nicotine (0.4 mg/kg per day, 6 days of treatment), when compared to rats exposed to saline. Reduction in immunoreactivity was significant for NF-68 (P < 0.05), NF-160 (P < 0.01) and NF-200 (P < 0.05), showing a relative reduction of 34%, 42% and 38%, respectively, when compared to saline-treated rats. No difference was observed for any of the NF under study when immunoreactivity measurements in the substantia nigra were compared. Ultrastructural analysis was applied to evaluate changes in NF-68, NF-160 and NF-200 immunoreactivity in regions of the VTA that contain dopaminergic neurons following chronic nicotine treatment. At the electron microscopic level, no degenerative changes were found in neurons or glial cells of the VTA. With ultrastructural immunohistochemistry, evaluation of the homogeneity parameter of NF distribution showed a loss of homogeneity for NF-68 linked to the nicotine treatment. In areas in which NF organization appeared well preserved, analysis of the numerical density of NF revealed no significant difference for NF-68 (897/ micro m2 vs. 990/ micro m2), NF-160 (970/ micro m2 vs. 820/ micro m2) and NF-200 (1107/ micro m2 vs. 905/ micro m2) in nicotine-treated rats when compared to saline-treated rats. These results confirm that nicotine shares the same properties with cocaine and morphine in reducing NF in the VTA, a key brain structure of the rewards system, and that chronic nicotine treatment changes the axonal distribution of 68 kDa neurofilaments in the rat VTA.

Human chorionic gonadotropin in human placentas from normal and preeclamptic pregnancies

The aim of this study was to examine the distribution of human chorionic gonadotropin (hCG) in the human placenta of normotensive and preeclamptic pregnancies and to determine by computer image analysis, whether differences in hCG immunoreactivity occurred in preeclamptic as apposed to normotensive pregnancies. We discuss how far elevated maternal serum levels of hCG normally observed in preeclamptic patients reflect an increased secretory activity of the syncytiotrophoblast. We used the immunoperoxidase technique to locate hCG. Quantification of immunostaining intensity was done by computer image analysis. In normotensive placentas from all the gestational ages human chorionic gonadotrophin immunoreactivity was specifically detected in the syncytiotrophoblast. There is an apparent decrease in the intensity of the hCG immunostaining in the syncytiotrophoblast from the 29th to 36th week of gestation in normotensive placentas. No hCG immunostaining was observed in the villous or extravillous cytotrophoblast of all placentas. In preeclamptic placentas the expression of hCG was homogeneous with a moderate to intense immunoreactivity in the syncytiotrophoblast. Microdensitometric analysis of the section from normotensive and preeclamptic placentas indicated that there is a statistically significant preeclampsia-induced increase in immunohistochemical reaction intensity for hCG (p < 0.05). This study seems to demonstrate that increased production of hCG by preeclamptic placentas is associated with strong hCG immunostaining of the syncytiotrophoblast.

Distant microglial and astroglial activation secondary to experimental spinal cord lesion.

This paper analysed whether glial responses following a spinal cord lesion is restricted to a scar formation close to the wound or they might be also related to widespread paracrine trophic events in the entire cord. Spinal cord hemitransection was performed in adult rats at the thoracic level. Seven days and three months later the spinal cords were removed and submitted to immunohistochemistry of glial fibrillary acidic protein (GFAP) and OX42, markers for astrocytes and microglia, as well as of basic fibroblast growth factor (bFGF), an astroglial neurotrophic factor. Computer assisted image analysis was employed in the quantification of the immunoreactivity changes. At the lesion site an increased number of GFAP positive astrocytes and OX42 positive phagocytic cells characterized a dense scar formation by seven days, which was further augmented after three months. Morphometric analysis of the area and microdensitometric analysis of the intensity of the GFAP and OX42 immunoreactivities showed reactive astrocytes and microglia in the entire spinal cord white and gray matters 7 days and 3 months after surgery. Double immunofluorescence demonstrated increased bFGF immunostaining in reactive astrocytes. The results indicated that glial reaction close to an injury site of the spinal cord is related to wounding and repair events. Although gliosis constitutes a barrier to axonal regeneration, glial activation far from the lesion may contribute to neuronal trophism and plasticity in the lesioned spinal cord favoring neuronal maintenance and fiber outgrowth.

Experimental Models of Partial Lesion of Rat Spinal Cord to Investigate Neurodegeneration, Glial Activation, and Behavior Impairments

The article demonstrates two experimental models of spinal cord partial injury in rats: a contuse model promoted by the NYU impactor system and a partial hemitransection model achieved by a stereotaxic-posi-tioned adjustable wire knife. By means of a defined impact weight (10 g) and a digital optical potentiometer linked to a computer, the impactor transferred and registered a moderate or a severe contusion to the rat spinal cord at a low thoracic level after dropping the weight from distances of 25 mm and 50 mm, respectively, to the dorsal surface of the exposed dura spinal cord. Impact curve was calculated and the parameters of the trauma, like impact velocity, cord compression distance and cord compression rates were obtained in order to demonstrate trauma severity. To promote partial hemitransection, rats were positioned in a spinal cord unit of a stereotaxic apparatus and lesion was made with the adjustable wire knife spatially oriented. By means of a computerized infrared motion sensor-home cage activity monitor and a noncomputerized evaluation of motor behavior using the inclined plane and the motor score of Tarlov tests, behavior was analyzed in an acute period postlesion. Rats were sacrificed and spinal cords were processed for routine staining to show neurons and for GFAP and OX42 immunohistochemistry to demonstrate glial cells. The tissue labelings were quantified using computer assisted stereology by means of an optical disector and microden-sitometric image analysis by means of quantification of gray values of discriminated profiles. While partial hemitransection model favored a more accurate control of the lesion location, the contuse model allowed us to perform different degrees of lesion severity. A close correlation between behavioral impairment and severity of trauma was seen in the rats submitted to spinal cord contusion. The stereologic lesion index showed a correlation between severity of trauma and tissue damage by 7 days and demonstrated a time-dependent secondary degeneration after moderate but not after severe spinal cord contusion from 7 to 30 days after injury. Long-lasting activations of astrocytes and microglia seen by persisted increases in the specific mean gray values of immu-noreactivities were also found in all levels of the white and gray matters of the partial hemitransected spinal cord until 3 months postinjury which can he related to wound/repair events and paracrine trophic support to spinal cord remaining neurons. The results showed that controlled partial lesions may provide an important toll to study trophism and plasticity in the spinal cord.

Fixation of the craniofacial skeleton with butyl-2-cyanoacrylate and its effects on histotoxicity and healing.

Butyl-2-cyanoacrylate is an easily applied, biocompatible, bioresorbable polymer glue that provides an alternative to conventional rigid fixation techniques. Our aim was to determine if cyanoacrylate fixation of the bone flap in a rabbit craniotomy model provides the healing and strength afforded by plate and screw fixation. We also investigated the inflammatory responses of adjacent tissues including the scalp, cranium, and brain. A unilateral parietal bone flap was elevated in 33 adult New Zealand rabbits. The bone was fixed in position with cyanoacrylate (n = 13), fixed with a microplate and screws (n = 14), or was replaced without fixation (sham-control, n = 6). Normal scar formation and no residual polymer were found in scalp specimens. Neuropathologic analysis identified the presence of residual polymer on the surface of 2 of the 13 rabbit brains. Histopathologic analysis of the bone flap-to-skull interface revealed no difference in the degree but rather in the quality of inflammation and healing between the plate and screw and polymer fixation groups. Microdensitometric analysis of the bone gap revealed nearly equivalent bone density in the cyanoacrylate and plated groups, tending to less density in the sham group (p = 0.11 and 0.09, respectively). An additional study focusing on neurotoxicity was performed in 20 adult rabbits with 3-week and 11-week recovery periods and similarly found the absence of a marked inflammatory response to the polymer. In conclusion, bone healing and soft-tissue inflammation were comparable between cyanoacrylate and plate and screw fixation groups. Although butyl-2-cyanoacrylate glue fixation may provide a reasonable alternative to hardware fixation, further investigations are necessary to identify its ideal utilization.

Immunohistochemical demonstration of beta-naphthoflavone-inducible cytochrome P450 1A1/1A2 in rat intrahepatic biliary epithelial cells.

Although intrahepatic biliary epithelial cells are targets for certain hepatotoxic chemicals, including some procarcinogens, their ability to monooxygenate, and thereby bioactivate and inactivate xenobiotics, remains to be established. Thus, the present study was undertaken to immunohistochemically determine if cytochrome P450 (CYP) 1A1/1A2 is present and can be induced within these nonparenchymal liver cells. Immunoperoxidase and immunofluorescent staining for CYP1A1/1A2 was detected within intrahepatic biliary epithelial cells as well as hepatocytes of control rats and was markedly enhanced in both cell types by beta-naphthoflavone (BNF). Color confocal laser microscopic analyses of dual immunofluorescent staining for CYP1A1/1A2 and cytokeratins 6 and 9 (56 and 64 kd, respectively) provided unequivocal evidence for the presence and induction of CYP1A1/1A2 within intrahepatic bile duct epithelia. Moreover, microdensitometric analyses of immunoperoxidase staining intensities for CYP1A1/1A2 revealed that intrahepatic biliary epithelial cells of control rats contain 44%, 56%, and 58% as much CYP1A1/1A2 as do centrilobular, midzonal, and periportal hepatocytes, respectively. These analyses further revealed that BNF increased the content of CYP1A1/1A2 in biliary epithelial cells by approximately 120%, while CYP1A1/1A2 levels in centrilobular, midzonal, and periportal hepatocytes were increased by 82%, 159%, and 160%, respectively. The results of this study represent the first in situ demonstration that mammalian intrahepatic biliary epithelial cells contain a CYP isoform, and further that CYP1A1/1A2 can be induced in these cells by BNF. These findings therefore indicate that intrahepatic biliary epithelial cells can oxidatively metabolize xenobiotics in situ and that their ability to bioactivate and inactivate xenobiotics can be significantly enhanced by CYP1A1/1A2 induction.

Age-related changes of sulphide-silver staining in the rat hippocampus

The influence of ageing on sulphide-silver positive zinc stores was assessed in the stratum radiatum of the CA 1–CA 3 sub fields of the rat hippocampus and in the molecular layer of the dentate gyrus using a silver amplification histochemical technique associated with microdensitometry. The volume of areas examined for microdensitometry was evaluated as well by quantitative image analysis. Male Sprague-Dawley rats aged 3 months (considered to be young), 12 months (considered to be adult) and 24 months (considered to be old) were used. Microdensitometric analysis of values of sulphide-silver staining corrected for the volume of hippocampal areas investigated revealed no age-dependent changes of staining in the CA 1 sub field of the hippocampus. In the CA 2 sub field a decrease of sulphide-silver staining was noticeable in aged rats in comparison with younger cohorts. A progressive reduction in the intensity of sulphide-silver staining was observed in the CA 3 sub field of the hippocampus. In the molecular layer of the dentate gyrus, the intensity of staining was decreased in adult and old rats in comparison with young animals. These findings indicate a different sensitivity to ageing of histochemically detectable zinc stores of rat hippocampus. The possibility of a specific sensitivity to senescence of different zinc-containing pathways of the hippocampus is discussed.

AD-like tau phosphorylation in human neuroblastoma cells following PKC hyperactivation is mediated by MAP kinase

SH-SY-5Y human neuroblastoma cells were treated with the phorbol ester TPA and the MAP kinase inhibitors PD98059. Microdensitometric analyses revealed that TPA treatment induced 60% and 46% increases in immunoreactivity towards monoclonal antibodies (AT-8 and PHF-1, respectively; p=0.0005) that recognize tau epitopes common to paired helical filaments. These increases were attenuated by the simultaneous treatment with PD98059. Steady-state AT-8 and PHF-1 immunoreactivity were not inhibited by PD98059. These analyses corroborate studies from other laboratories that indicate that MAP kinase may not normally confer AD-like immunoreactivity to tau within intact cells, but demonstrate that hyperactivation of upstream kinase(s) may recruit otherwise benign kinases to hyperphosphorylate tau. These findings further implicate the potential involvement of hyperactivation of signal transduction pathways in the early events that propagate PHF formation. © 1997 John Wiley & Sons, Ltd.

Variation in Genome Size in Benthic Polychaetes: Systematic and Ecological Relationships

Genome size (or C-value) in benthic polychaetes was studied from both experimental data and literature. Nuclear DNA content variation was analysed as related to both phylogenetic relationships of various families, general life habit of the species (interstitial vs macrobenthic), diploid chromosome number (2n), and mean body size (length). Experimental data on genome size were obtained for 43 lower taxa (genera and species) by means of microdensitometric analysis of the amount of Fuelgen-stained haploid DNA in interphase nuclei. These data, added to those available in the literature (36 taxa studied by Conner et al., 1972) gave a total of 79 taxa (genera and species), representing 30 families and 14 orders. Twenty-one of the species studied belonged exclusively to interstitial (=meiofaunal) families or genera.