Oval Morphology Research Articles

Characterization of a Cryptosporidium muris infection and reinfection in CF-1 mice

To establish control values for circulating cells and immune associated organs over the course of a self-limiting Cryptosporidium muris infection and rechallenge infection, mice were sacrificed at intervals starting before oral inoculation and ending after oocyst shedding had ceased. These values were used in other experiments to evaluate changes in these parameters induced by a single dose glucocorticoid immunosuppression model and in other immunosuppression studies. Flow cytometry counts of circulating T-lymphocytes and neutrophils, differential leukocyte counts, leukocyte morphology, spleen and thymus changes, and oocyst shedding were evaluated. Immediately after C. muris oocyst inoculation and up to the start of oocyst production (day 0 to day 7), the circulating blood profile showed a 50% drop in all leukocytes, including both large and small lymphocytes and CD3, CD4 and CD8 T-lymphocytes. There was an initial slight rise in circulating mature neutrophils after oocyst inoculation but numbers promptly dropped below normal and remaineded below normal. In the differential cell counts, monocytes with a fat, oval morphology increased by 60% at 24 h and remained high through oocyst shedding and beyond (day 8 through day 36). During oocyst shedding and continuing past the end of shedding, T-lymphocytes increased 100%. Monocytes with a flat, angular morphology increased in a similar manner. Immediately after oocyst inoculation the spleen contracted by 29%, but became 92% larger than its pre-inoculation size by day 14 when heavy oocyst shedding began. It remained enlarged through the end of oocyst shedding (day 29) and beyond (day 36). Spleen volume decreased and increased similar to changes in T-cell numbers. Throughout the C. muris infection the thymus remained largely unchanged. The transit of an oocyst bolus was followed from the stomach through the gut to the colon. No oocysts could be found in the stomach, caecum or feces of mice one half hour after oocyst inoculation. Likewise, an oral bolus of India ink passed from the stomach entirely into the colon after 3 h; therefore, no oocysts from the inoculum passed completely through the intestine and out into the feces. Recovered mice rechallenged with C. muris showed increased B-lymphocyte numbers; however, T-lymphocyte numbers remained level. The large lymphocytes increased after rechallenge, peaking on day 3, then decreased through day 10. B-cell numbers followed a pattern similar to the large lymphocytes. On day 10 of infection monocytes with a fat oval morphology rose sharply while B-cells fell in number. In both the initial infection and the rechallenge there was no unique blood profile which could definitely indicate a protozoal disease or identify a specific point during the course of the disease. There was no increase in the number of either small or large lymphocytes prior to increases in fat or flat monocytes.

The prevalence of oval root canals in the apical and middle third of mandibular molars and premolars

Introduction. Oval root canals are all groups of human teeth. In such canals it is a challenge to fulfill the necessary preconditions for a successful outcome of root canal therapy - complete cleaning, shaping and obturation. Aim. The aim of this study was to investigate the shape and diameter of root canals in the apical and middle third of human mandibular molars and premolars, to determine the prevalence and extent of long oval canals. Materials and methods. The study was carried out on 80 extracted human teeth, 40 molars and 40 premolars. The teeth were placed in a specially designed muffle mould and embedded in transparent acrylic resin. After the polymerization of the acrylic resin, the teeth were taken out of the mould and sectioned at levels 3 and 6 mm from the apex. The cross-sections were photographed under 30? magnification using a digital camera. The long and short canal diameter were measured on the images and their ratio calculated. Results. In the apical third 22% of the premolars and 35% of the molars had oval root canal morphology. In the middle third 36% of premolars and 41 % of molars. Conclusion. Root canals of mandibular molars and premolars are often oval in their cross-section. They seem to be more frequently oval in their cross-section than previous studies have shown.

Glucose-regulated Glucagon Secretion Requires Insulin Receptor Expression in Pancreatic α-Cells

The insulin receptor (IR) and its signaling appear to be essential for insulin secretion from pancreatic beta-cells. However, much less is known about the role of the IR in alpha-cells. To assess the role of the IR in glucagon and insulin secretion, we engineered adeno-viruses for high efficiency small interference RNA (siRNA)-IR expression in isolated mouse pancreatic islets and lentiviruses for siRNA-IR expression in pancreatic alpha- and beta-cell lines (alpha-TC6 and MIN6) with specific, long term stable IR knockdown. Western blot analysis showed that these strategies resulted in 60-80% reduction of IR protein in islets and alpha- and beta-cell lines. Cell growth was reduced by 35-50% in alpha-TC and MIN6 cells stably expressing siRNA-IR, respectively. Importantly, glucagon secretion, in response to glucose (25 to 2.8 mm), was completely abolished in islets expressing siRNA-IR, whereas secretion increased 1.7-fold in islets expressing control siRNA. In contrast, there was no difference in glucose-stimulated insulin secretion when comparing siRNA-IR and siRNA control, with both groups showing a 1.7-fold increase. Islet glucagon and insulin content were also unaffected by IR knockdown. To further explore the role of the IR, siRNA-IR was stably expressed in pancreatic cell lines, which dramatically suppressed glucose-regulated glucagon secretion in alpha-TC6 cells (3.4-fold) but did not affect GSIS in MIN6 cells. Defects in siRNA-IR-expressing alpha-cells were associated with an alteration in the activity of Akt and p70S6K where insulin-induced phosphorylation of protein kinase B/AKt was greatly reduced while p70S6K activation was enhanced, suggesting that the related pathways play important roles in alpha cell function. This study provides direct evidence that appropriate expression of the IR in alpha-cells is required for glucose-dependent glucagon secretion.

Comparative spatial and temporal localisation of perlecan, aggrecan and type I, II and IV collagen in the ovine meniscus: an ageing study

This is the first study to immunolocalise perlecan in meniscal tissues and to demonstrate how its localisation varied with ageing relative to aggrecan and type I, II and IV collagen. Perlecan was present in the middle and inner meniscal zones where it was expressed by cells of an oval or rounded morphology. Unlike the other components visualised in this study, perlecan was strongly cell associated and its levels fell significantly with age onset and cell number decline. The peripheral outer meniscal zones displayed very little perlecan staining other than in small blood vessels. Picrosirius red staining viewed under polarised light strongly delineated complex arrangements of slender discrete randomly oriented collagen fibre bundles as well as transverse, thick, strongly oriented, collagen tie bundles in the middle and outer meniscal zones. The collagen fibres demarcated areas of the meniscus which were rich in anionic toluidine blue positive proteoglycans; immunolocalisations confirmed the presence of aggrecan and perlecan. When meniscal sections were examined macroscopically, type II collagen localisation in the inner meniscal zone was readily evident in the 2- to 7-day-old specimens; this became more disperse in the older meniscal specimens. Type I collagen had a widespread distribution in all meniscal zones at all time points. Type IV collagen was strongly associated with blood vessels in the 2- to 7-day-old meniscal specimens but was virtually undetectable at the later time points (>7 month).

Patent Foramen Ovale Morphology and Impact on Percutaneous Device Closure

We sought to evaluate the effects of atrial septal and patent foramen ovale (PFO) morphology on the efficacy of transcatheter closure. We performed a retrospective analysis of all patients who underwent PFO device closure from January 1997 to January 2002. Forty-seven patients underwent percutaneous closure of a PFO with a median age of 45 years (range, 8-75) and weight of 76 kg (range 28-115). The septal morphology was flat in 33 and aneurysmal in 14 patients. The PFO morphology was a simple flap in 20 and complex in 27 patients. Complex morphologies included long-tunnel PFO (n = 15), coexistent small atrial septal defect (ASD) (n = 5), and aneurysmal septum without a tunnel or ASD (n = 7). Nonstretched PFO diameters were significantly smaller than stretched (4.8 +/- 1.1 mm vs 11.6 +/- 3.8 mm, p < 0.01). Median device size: stretched diameter ratio (DS:SD) was 3.7:1 (range, 2.2-9.1). The DS:SD ratio was significantly higher in patients with complex PFO (mean, 3.9:1 vs 2.6:1; p < 0.05). Device placement was successful in all patients. Five patients required transeptal puncture of the foraminal flap in long-tunnel PFOs. Effective closure on follow-up was achieved in 45 patients (95%). Of the 2 patients with residual shunts, 1 had a complex PFO and the other a simple PFO. Two patients (5%) experienced recurrent neurologic symptoms after device closure despite having no residual shunt by echocardiography. Complex PFO did not increase risk of residual shunt or recurrent neurologic symptoms after device closure.

Swelling behavior and cell viability of dehydrothermally crosslinked poly(vinyl alcohol) hydrogel grafted with N‐vinyl pyrrolidone or acrylic acid using γ‐radiation

AbstractTo improve equilibrium water content, dehydrothermally crosslinked poly(vinyl alcohol) (PVA) hydrogel was grafted with N‐vinyl pyrrolidone (NVP) or acrylic acid (AA) monomer using γ‐radiation. Swelling behavior of the grafted hydrogels was studied in phosphate‐buffered saline, and cell viability was evaluated using fibroblast cells from mouse connective tissue. Equilibrium water content of AA‐ and NVP‐grafted PVA hydrogel ranged between 40–60% and 60–80%, respectively, depending on radiation dose and monomer concentration. For maximum degree of swelling, the optimum monomer concentration and radiation dose were 20% by weight and 20 kGy, respectively. Fibroblast cells seeded on NVP‐grafted hydrogel had an extended oval morphology while those seeded on AA‐grafted PVA had a rounded spherical morphology. These results support the use of NVP for grafting PVA to increase swelling and improve cell viability. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2862–2868, 2004

An investigation on the microstructure of an AM50 magnesium alloy

The microstructure and the dislocation arrangement in the die cast AM50 magnesium alloy as well as in the stressed states have been investigated using conventional transmission electron microscopy (TEM), high-resolution TEM and energy dispersive X-ray analysis. The microstructure of the die cast AM50 alloy is found to mainly consist of α-Mg, β-Mg 17Al 12 and Al 8Mn 5 phases. Two kinds of β-Mg 17Al 12 particles with different sizes have been found in the stressed AM50 magnesium alloy. Besides the normal β-Mg 17Al 12 particles with size around several micrometers as in the die-cast AM50 magnesium alloy, some finer β-Mg 17Al 12 particles in the stressed states of the AM50 alloy have also been found. The finer β-Mg 17Al 12 particles are found to be only about tens of nanometers with oval or rod-like morphology, which may be formed during deformation. Dislocation pile-ups have been found in the stressed AM50 alloy for the first time. The spacing between each parallel dislocation in the pile-ups is only several nanometers. The dislocations are confined in the slip planes and piled up against grain boundaries. Dislocations in the networks are found to increase with deformation of the alloy. Also, dislocation networks have been found in the β-Mg 17Al 12 and Al 8Mn 5 phases as well as in the matrix in the deformed AM50 magnesium alloy. However, the dislocation pile-ups are found to be almost identical from 1.3% deformation to rupture, which explains the stable tensile yield strength of the AM50 magnesium alloy during the deformation.

Wide distribution of Plasmodium ovale in Myanmar.

The presence of Plasmodium ovale has never been previously reported in Myanmar. Using blood samples obtained in many villages across the country between 1996 and 2000, molecular diagnosis of Plasmodium species was made with semi- or full-nested polymerase chain reaction (PCR) with species-specific primers, followed by agarose gel electrophoresis to detect amplification products. The presence of P. ovale was also confirmed with the another PCR-based diagnosis, the microtiterplate hybridization (MPH) method using species-specific probes. Both methods target the A type of the small subunit ribosomal RNA gene of the four human malaria parasites. Plasmodium ovale DNA was amplified in samples from 65 (4.9%) of 1323 PCR-positive patients, with perfect agreement between results obtained by nested PCR and MPH. Only four P. ovale-infected patients had single-species infection; all others were coinfected with P. falciparum, P. vivax and/or P. malariae. Quadruple infections were observed in six subjects. Parasites with typical P. ovale morphology were found in only 19 patients by conventional microscopy of Giemsa-stained thin smears or fluorescence microscopy of acridine orange-stained thin smears. Plasmodium ovale infections were found in villages situated in the southern, central and western regions of Myanmar, suggesting that P. ovale may be widely distributed in this country.

Imported Plasmodium vivax malaria in France: geographical origin and report of an atypical case acquired in Central or Western Africa

A total of 73 cases of Plasmodium vivax infections acquired in Western or Central Africa were diagnosed on microscopical criteria in French travellers from 1995 to 1998. We report a case of P. vivax infection in a non immune traveller confirmed by polymerase chain reaction and presenting an atypical P. ovale morphology. The infection was acquired in Western or Central Africa. These microscopical observations, together with the molecular evidence for P. vivax in Western and Central Africa suggest that P. vivax is transmitted in this area despite lacking the Duffy receptor in autochthonous population.

Measurement of ovalocyte frequency in peripheral blood smears in defining ovalocytosis in Papua New Guinea.

Red cell oval morphology is still the only accepted basis for the clinical or epidemiological diagnosis of ovalocytosis. Therefore it is important to know the errors when detecting and counting morphological ovalocytes. In all previous studies of ovalocytosis there was no assessment of the variation which may have occurred in classification due to smearing and staining techniques or the criteria for the diagnosis of ovalocyte morphology; nor was inter or intraobserver variation assessed. We report how different peripheral blood smear methods influence the diagnosis of ovalocytosis in populations in the Madang and East Sepik Provinces in Papua New Guinea. We also examined within and between observer variation in the quantitative assessment of ovalocytosis at x 40 and x 100 microscopy powers. A modified method of making a thin malaria blood smear gave the best preservation of red cell morphology and was adopted for the quantitative ovalocytosis studies. A special haematology smear is unnecessary. Ovalocyte frequency estimations were similar when x 40 and x 100 lenses were used, but x 40 was preferable for assessing morphology. Two observers were consistent in their findings and produced very similar results for the high-quality smears from the planned Madang survey, and rather different results for the smears from the unplanned routine Sepik survey. We conclude that measurement error for ovalocytosis assessment can be quite small and unimportant, minimized by careful planning and quality control. Otherwise measurement error is substantial and threatens validity of classification and grading of ovalocytosis.

Humoral immune response to Malassezia furfur in patients with pityriasis versicolor and seborrheic dermatitis.

Humoral immune responses against exoantigen components of oval, elliptic and round yeast forms of Malassezia furfur were analysed by ELISA and Western blotting assays, using sera from patients with pityriasis versicolor (PV), seborrheic dermatitis (SD) and healthy adults (HA), as control. Sera from patients with SD showed IgG anti-oval M. furfur titers ranging from 1/400 to 1/6400 showing geometric mean (GM) of 1/1472, higher than those obtained with sera from patients with PV (1/200 to 1/6400, GM = 1/1239). Both patient groups showed mean titres statistically superior (P < 0.05) than those obtained form HA (GM = 1/229). Similar data were also obtained with the elliptic and round antigens. However, the anti-oval IgG mean titers from patients' sera were much higher than those obtained with elliptic or round antigenic components (p < 0.05) Anti-M furfur IgM titers obtained from patient's sera with PV against all three exoantigens were statistically superior (p < 0.05) than HA group. Patients with SD showed IgM titers statistically superior (p < 0.05) only to oval yeasts of M. furfur. The IgA mean titers from patients' groups against the different morphological antigens were shown be slightly higher than those HA group. By Western blot, using rabbit anti-sera, the different antigenic components of M.furfur showed a close relationship mainly between oval and elliptic yeast cells antigens. The 70 kDa component of the M. furfur exoantigen of oval morphology was recognized by 84% of the PV patients' sera. On the other hand, SD patients' sera recognized 3 principal components of 70 kDa (100%), 65 kDa (67%) and 84 kDa (53%). These components may be considered immunological markers for PV and SD. Twenty-five percent of HA sera recognized the components of 65, 70 and 94 kDa. This investigation shows that M. furfur antigens can sensitize the host, mainly the oval yeast form of M. furfur with a very important specific IgG response in patients with SD and PV.

The double oval UV auroral distribution: 1. Implications for the mapping of auroral arcs

During the later stages of the auroral substorm the luminosity distribution frequently resembles a double oval, one oval lying poleward of the normal or main UV auroral oval. We interpret the double oval morphology as being due to the plasma sheet boundary layer becoming active in the later stages of the substorm process. If the disturbance engulfs the nightside low‐latitude boundary layers, then the double oval configuration extends into the dayside ionospheric region. The main UV oval is associated with the inner portion of the central plasma sheet and can rapidly change its auroral character from being diffuse to discrete. This transition is associated with the substorm process and is fundamental to understanding the near‐Earth character of substorm onset. On the other hand, the poleward arc system in the nightside ionosphere occurs adjacent to or near the open‐closed field line boundary. This system activates at the end of the optical expansion phase and is a part of the recovery phase configuration in substorms where it occurs. These two source regions for nightside discrete auroral arcs are important in resolving the controversy concerning the mapping of arcs to the magnetosphere. The dayside extension of this double oval configuration is also investigated and shows particle signatures which differ considerably from those on the nightside giving clues to the magnetospheric source regions of the aurora in the two local time sectors. Near‐Earth substorm onsets are shown to be coupled to processes occurring much further tailward and indicate the importance of understanding the temporal development of features within the double oval. Using “variance images,” a new technique for the investigation of these dynamics is outlined.

Effects of four methods of sperm preparation on the motile concentration, morphology, and acrosome status of recovered sperm from normal semen samples.

This prospective study compared the effects of four sperm preparation methods on semen samples with normal sperm parameters. The samples were obtained from husbands of infertile couples being evaluated at a fertility center in New York City. Twenty-nine of 51 men in the study (56.9%) had fathered a child within the past 2-5 years. Specifically, the study compared the effects of simple wash (unselective) and three selective methods--conventional swim-up, three-gradient Percoll separation, and swim-up from the Percoll-separated fraction--on the concentration, percentage motile, percentage normal (oval) morphology, percentage intact acrosomes, percentage motile sperm recovery, and motile sperm concentration in the processed samples. The same parameters were measured in the untreated ejaculate, resulting in five data sets. These data were analyzed by both comparison across the five sets and pairwise multiple comparisons between sets. The study showed that of the selective methods, conventional swim-up and Percoll separation yielded comparably optimal percentages motile sperm recovery and motile sperm concentrations. However, swim-up yielded a significantly higher percentage oval and percentage motile sperm than Percoll, while Percoll had a significantly higher percentage acrosome-intact. Swim-up after Percoll separation yielded the highest percentage motility and percentage oval but had the lowest motile sperm concentration with the lowest percentage acrosome-intact.

Ultrastructural study of cholinergic neurons in the medial septal nucleus and vertical limb of the diagonal band of broca in the basal forebrain of the rat

The morphology, ultrastructure and synaptic relationships of the cholinergic and non-cholinergic neurons in the medial septal nucleus (MS) and vertical limb of the diagonal band of Broca (VDB) in the basal forebrain of the rat were studied at the light and electron microscopic levels. The cholinergic neurons were localized immunocytochemically using a monoclonal antibody against choline acetyl-transferase (ChAT). Morphometric and statistical analyses showed that ChAT-labelled cells presented a predominantly oval morphology in both nuclei. The sizes of the neurons were significantly larger in the VDB nucleus. Within the two nuclei, two populations of cholinergic neurons were differentiated. One of the large immunolabelled neurons presented deep indentations and prominent nucleoli in their non-immunoreactive nuclei. Their cytoplasm contained a well-organized endomembrane system composed of short cisternae of rough endoplasmic reticulum (RER). One or two lamellar bodies with a peculiar ultrastructure were frequently found intercalated in this system. The Golgi areas presented numerous coated vesicles, sequestration and multivesicular bodies, which was indicative of an intense metabolic activity in these cells. The second population of small immunolabelled neurons exhibited reduced cytoplasm with a poorly developed endomembrane system and apparent absence of lamellar bodies. The neighbouring non-immunolabelled neurons presented a different type of organization of the endomembrane system which was composed of scattered and loosely arranged elongated cisternae of RER and infrequent lamellar bodies, with a structure different from that seen in the large cholinergic neurons. We propose that the structural differences in composition of the endomembrane system and lamellar bodies observed in the three types of neurons in this study indicate different metabolic activities. Symmetrical and asymmetrical synaptic contacts were observed on somata and dendrites of labelled neurons, the latter being more frequent. ChAT-labelled axon boutons were never seen. The absence of immunolabelled axon terminals and the presence of immunolabelled myelinated axons leads us to suggest that the majority of neurons in these areas are of the long projecting type.

Histopathology and cell culture characteristics of liver cells from grc − and grc + rats given diethylnitrosamine

The histopathological response and cell culture characteristics of liver cells from the R16 (grc-) strain of rats, which carries an MHC-linked deletion, were examined one week after a single intraperitoneal injection of 200 mg/kg body weight diethylnitrosamine (DEN) and were compared with the response of liver cells from wild type (grc+) rats. The DEN exposure induced hydropic/vacuolar changes in the parenchymal cells and a limited proliferation of oval cells in the periportal areas of the livers of both grc+ and grc- rats. Primary culture of collagenase-digested livers consisted of parenchymal, bile ductular and oval-related cells as determined by cell-specific immunohistochemistry. Subpassaged cells from grc+ rats exhibited oval cell ultrastructural morphology, inducible histochemical staining for gamma-glutamyl transpeptidase (GGT), and DEN-associated onset of anchorage-independent growth. Primary cultures of liver cells from R16 rats consistently failed to form cell strains upon subpassage.

Sperm morphology assessment as an indicator of human fertilizing capacity.

Semen samples from 95 men were examined by routine semen analysis and specific histologic staining for sperm morphology. The men were classified into fertile and infertile groups on the basis of clinical evaluation and in vitro testing, using the zona-free hamster egg penetration assay. Thirty men were designated as fertile, as they had fathered children and their sperm showed penetration of greater than 20% of the zona-free hamster eggs with the in vitro fertilization test. Subjects classified as infertile were men from infertile couples whose wives showed no evidence of infertility and whose in vitro fertilization ability was 10% or less. The semen analysis parameters of the fertile and infertile groups were significantly different. Fertile men had mean values of 108 X 10(6) sperm/ml, 61% motile, 64% normal forms (sperm with oval morphology), and 69% penetration in vitro. The mean values for infertile men were significantly lower: 42 X 10(6) sperm/ml, 45% motile, 32% normal forms, and 3.2% penetration in vitro. The importance of the morphology parameter was revealed by comparison of the percentage of penetration with count, motility, and morphology. Penetration correlated best with morphology (r = 0.730) as compared with motility (r = 0.451) and count (r = 0.605). The distribution of abnormalities in the infertile group revealed 81.6% with abnormal morphology (less than 50%), 53.8% with abnormal motility (less than 50%), and 38.5% with abnormal count (less than 20 million/ml). As a single parameter, decreased number of normal forms appears to be a good indicator for clinical infertility if in vitro fertilization testing is not available.

The latitudinal morphology of 10-eV to 10-keV electron fluxes during magnetically quiet and disturbed times in the 2100-0300 MLT sector

Electron and proton data from Isis 1 and 2 have been used to examine the change in the latitudinal morphology of auroral particle fluxes as a function of substorm time in the dipole magnetic local time period from 2100 to 0300. Clear and repeatable systematics in the latitudinal morphology were observed during the various phases of a substorm, allowing the substorm phase to be identified on the basis of particle data alone. Based on this study, a new phenomenological substorm model is presented which relates particle precipitation from various parts of the magnetotail to auroral oval morphology.