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PubMed1998

Infectivity of Cryptosporidium muris isolated from cattle.

The infectivity of a bovine isolate of Cryptosporidium muris for various animals was studied by transmission experiments. Neonatal BALB/c mice, adult BALB/c mice, SCID mice, common voles (Microtus arvalis), bank voles (Clethrionomys glareolus), common field mice (Apodemus sylvaticus), Mongolian gerbils (Meriones unguiculatus), desert gerbils (Gerbilus gerbilus), guinea pigs, rats, rabbits and goats were used to test the infectivity of this isolate. Among these host species, only Mongolian gerbils were susceptible to the infection and discharged C. muris oocysts in their faeces. The prepatent period for 8-week-old Mongolian gerbils was 15-19 days, the patent period varied between 18 and 36 days. More protracted chronic infections have been observed in gerbils immunosuppressed with methylprednisolone. No signs of clinical illness or macroscopic changes were seen in infected gerbils. Cryptosporidial developmental stages were detected in the stomach, histopathological changes were characterized by epithelial hyperplasia and mucosal hypertrophy without inflammatory exudate. In spite of the fact that C. muris was able to infect gerbils, we do not consider gerbils to be a true hosts for C. muris of cattle origin. Based on our results, we suggest that significant differences in host specificity of individual C. muris isolates exist, and that wild rodents are not reservoir for C. muris infection of cattle.

Animals↗
PubMed1999

Experimental Cochlosoma anatis infections in poultry.

Cochlosoma anatis [Kotlán, A., 1923. Zentralbl. Bakteriol. Parasitenkd. Infektienskr. Hyg. 90, pp. 24-28] is a flagellated protozoan parasite of birds. We have encountered C. anatis in turkeys with enteritis. Experimental oral inoculations of turkeys with 1 x 10(6) to 10 x 10(6) trophozoites consistently reproduced infections in recipients. Trophozoites were most numerous in the jejunum and ileum but could be observed in the duodenum, ceca, colon, and feces. When 12 naive turkeys were placed on contaminated litter vacated by excreting turkeys only one of 12 became infected. When eight naive turkeys were placed in boxes with birds currently excreting trophozoites, seven of eight became infected. Trophozoites could not survive exposure to water or to freezing. Attempts to culture trophozoites in modified Diamond's medium, Kiester's medium, RPMI 1640 medium with 10% fetal bovine serum, or on cultured bovine turbinate cells were not successful. Four of six bobwhite quail and one of eight chickens orally inoculated with 10 x 10(6) to 20 x 10(6) trophozoites had detectable infections. Trophozoites were observed only in the ilea of bobwhite quail and the ceca of the positive chicken. Trophozoites collected from chickens and bobwhite quail remained infectious for turkeys.

Animals↗
PubMed2001

High resolution imaging of the distribution and permeability of methyl viologen dication in bovine articular cartilage using scanning electrochemical microscopy.

Scanning electrochemical microscopy (SECM) has been used in the induced transfer (SECMIT) mode to image the permeability of a probe cation, methyl viologen (MV(2+)), in samples of articular cartilage. An ultramicroelectrode (UME), scanned just above the surface of a sample, is used to amperometrically detect the probe solute. The resulting depletion of MV(2+) in solution induces the transfer of this cation from the sample into the solution for detection at the UME. The current provides quantitative information on local permeability, provided that the sample-UME distance is known. It is shown that the necessary topographical information can be obtained using the amperometric response for the oxidation of Ru(CN)(4-)(6), which does not permeate into the cartilage matrix. This procedure was validated by marking samples in situ, after electrochemical imaging, with subsequent examination by ex situ interferometry and optical microscopy. Wide variations in the permeability of MV(2+) have been detected by SECMIT. These observations represent the first demonstration of the inhomogeneous permeability of a cation in cartilage on a micrometre scale. The permeability maps show similar features to the proteoglycan distribution, identified by toluidine blue staining, and it is likely that proteoglycans are the main determinant of MV(2+) permeability in articular cartilage.

Animals↗
PubMed2001

Evidence of altered inhibition in layer V pyramidal neurons from neocortex of Kcna1-null mice.

Mice lacking the potassium channel subunit KCNA1 exhibit a severe epileptic phenotype beginning at an early postnatal age. The precise cellular physiological substrates for these seizures are unclear, as is the site of origin. Since KCNA1 mRNA in normal mice is expressed in the neocortex, we asked whether neurons in the neocortex of three to four week-old Kcna1-null mutants exhibit evidence of hyperexcitability. Layer V pyramidal neurons were directly visualized in brain slices with infrared differential-interference contrast microscopy and evaluated with cellular electrophysiological techniques. There were no significant differences in intrinsic membrane properties and action potential shape between Kcna1-null and wild-type mice, consistent with previous findings in hippocampal slice recordings. However, the frequency of spontaneous post-synaptic currents was significantly higher in Kcna1-null compared to wild-type mice. The frequency of spontaneous inhibitory post-synaptic currents and miniature (action-potential-independent) inhibitory post-synaptic currents was also significantly higher in Kcna1-null compared to wild-type mice. However, the frequency of spontaneous and miniature excitatory post-synaptic currents was not different in these two groups of animals. Comparison of the amplitude and kinetics of miniature inhibitory and excitatory post-synaptic currents revealed differences in amplitude, rise time and half-width between Kcna1-null and wild-type mice. Our data indicate that the inhibitory drive onto layer V pyramidal neurons is increased in Kcna1 knockout mice, either directly through an increased spontaneous release of GABA from presynaptic terminals contacting layer V pyramidal neurons, or an enhanced excitatory synaptic input to inhibitory interneurons.

Animals↗
PubMed2001

Basilar membrane vibration in the basal turn of the sensitive gerbil cochlea.

The basal membrane (BM) velocity responses to pure tones were measured using a newly developed laser interferometer microscope that does not require placing a reflecting object on the BM. It was demonstrated that the instrument is able to measure sub-nanometer vibration from the cochlear partition in the basal turn of the gerbil. The overall shape of the amplitude spectra shows typical tuning features. The 'best' frequencies (BFs) for the BM locations studied were between 14 kHz and 27 kHz, depending on the longitudinal position. For a given BM location, tuning sharpness was input level dependent, indicated by the Q(10dB), which varied from approximately 3 at low stimulus levels to near 1.5 at high input levels. At frequencies below BF, parallel amplitude/frequency curves across stimulus levels indicate a linear growth function. However, at frequencies near BF, the velocity increased linearly at low levels (<40 dB SPL) and became compressed between 40 and 50 dB SPL. Although the velocity gain for the frequency range below BF was a function of frequency, for a given frequency the gains were approximately constant across different levels. At frequencies near BF, the velocity gain at low sound pressure level was greater than that at a high sound pressure level, indicating a nonlinear negative relationship to stimulus level. The data also showed that the BF shifts toward the low frequencies with stimulus intensity increase. The phase spectra showed two important features: (1) at frequencies about half octave below the BF, phase slope is very small, indicating an extremely short delay; (2) the greatest phase lag occurs at frequencies near the BF, indicating a significant delay near this frequency range.

Acoustic Stimulation↗
PubMed1997

Firing properties of spherical bushy cells in the anteroventral cochlear nucleus of the gerbil.

In gerbils, spherical bushy cells (SBCs) encode low frequency sound signals into a temporal firing pattern. To investigate the support for the timing in this temporal code, we characterized the membrane electrical properties of visually identified SBCs in brainstem slices. A brief depolarizing subthreshold transient potential (TP) triggered, with relatively invariant latency, a single spike at the onset of a response to depolarizing current pulses. The activation of a subthreshold Na+-conductance, sensitive to blockade with tetrodotoxin, and a high threshold Ca2+-conductance, sensitive to blockade with Co2+ or Cd2+, accelerated the rising phase and amplified the TP. A K+-conductance, sensitive to blockade by 4-aminopyridine (4-AP, 50 microM), shaped the decay of the TP. Following a single spike, voltage-gated activation of transient and sustained K+-conductances suppressed any tendency to repetitively discharge. A reduction in either K+-conductance due to application of 4-AP or tetraethylammonium (TEA, 10 mM), converted the single spike mode to repetitive firing during the depolarizing pulses. A persistent, tetrodotoxin-sensitive Na+-conductance amplified steady-state depolarizing responses. A hyperpolarization-activated conductance, greatly decreased by extracellular Cs+ (3 mM) but resistant to Ba2+ (up to 1 mM), filtered the responses to hyperpolarizing current inputs. A depolarized membrane potential promoted repetitive firing in SBCs. This state, expected in pathophysiological conditions, would corrupt the temporal code.

4-Aminopyridine↗
PubMed2001

Examination of line crossings by atomic force microscopy.

Until now, the most widely used methods for the forensic examination of line crossings in documents were optical and electron microscopy. The combination of both techniques allows one in most cases to establish the sequence of lines. The recent development of scanning probe microscopy [1] gives an opportunity to complement or even replace the classical instruments used in this field. Scanning probe microscopes have been designed to study surfaces at high magnification. The aim of this study was to verify if their most popular member, the atomic force microscope (AFM) [2], can be applied to line crossing problems. The results show for the first time that AFM images present the same qualitative information obtained by scanning electron microscope (SEM) images and, consequently, allow the determination of the line crossing sequence under ambient conditions without vacuum and conductive coating of specimens.

Forensic Medicine↗
PubMed2004

In vivo surface wear mechanisms of femoral components of cemented total hip arthroplasties: the influence of wear mechanism on clinical outcome.

The appearance and mechanism of femoral stem wear was studied in 172 retrieved femoral components, of which 74 stems had been stable in vivo. Macroscopic, microscopic, and nano-level scales of examination were used. Loss of stem surface in response to micromotion (wear) was found to affect 93% of stems. However, changes were frequently difficult to see with the naked eye, and in 19% of cases they would have been missed completely without the use of light microscopy. The surface finish of the prosthesis determined the mechanism of stem wear. Matte surfaces showed typical abrasive processes that also damage the cement, releasing particulate debris from the cement and metal surfaces. This may destabilize the stem within the cement. Polished stems showed a typical fretting appearance with retention of debris on the stem surface and without significant damage to the cement. These differences in wear mechanism between matte and polished stems have significant effects on stem function.

Arthroplasty, Replacement, Hip↗
PubMed1999

Evaluation of an unused 1952 Ridley intraocular lens.

PURPOSE: To evaluate an unused 1952 historic Ridley intraocular lens (IOL) brought to Bombay, India, in 1952 from an Oxford Ophthalmologic Conference in England and given to 1 of the authors during his residency. SETTING: Alcon Laboratories, Fort Worth, Texas, USA. METHODS: The Ridley IOL was evaluated at Alcon Laboratories, Inc., using the established procedures of its Intraocular R&D Laboratories. Various optical and physical aspects of the Ridley lens were evaluated including (1) dimensions, (2) weight, (3) power, (4) resolution efficiency and modulation transfer function (MTF), (5) surface sphericity by interferometry, (6) ultraviolet (UV)-visible transmission characteristic, (7) attenuated total reflectance (ATR)-Fourier transform infrared reflectance spectrum, and (8) cosmetics by visual inspection using light microscopy. RESULTS: This 8.5 mm diameter, 2.4 mm thick, 23 diopter biconvex IOL weighed 108 mg. The ATR spectrum, UV-visible transmission, and refractive index confirmed its poly-(methyl methacrylate) material. The 0.56 MTF value at 100 line pairs/mm, per the International Standards Organization--IOL Optics Standard, and 93% resolution efficiency in water, per the American National Standard Institute IOL Optics Standard, revealed the IOL's excellent optics. This was confirmed by 0.278 wave root mean square surface figure as measured by Zygo interferometer using a 633 nm wavelength. Visual inspection revealed rough edges with sharp corners and some surface scratches. Early clinical experience with Ridley IOLs in Bombay, India, is briefly given. CONCLUSION: The Ridley IOL had excellent optical quality, meeting the requirements of current IOL optics standards. The selection of its dimensions was guided by the human crystalline lens, and the Ridley IOL was half as bulky. Although its clinical results were mixed, successful cases inspired subsequent improvements, leading to modern, highly satisfactory IOLs. This IOL represented a revolutionary innovation in ophthalmology.

England↗
PubMed2003

Elm1p is one of three upstream kinases for the Saccharomyces cerevisiae SNF1 complex.

BACKGROUND: The yeast SNF1 protein kinase and the mammalian AMP-activated protein kinase are highly conserved heterotrimeric complexes that are "metabolic master switches" involved in the switch from fermentative/anaerobic to oxidative metabolism. They are activated by cellular stresses that deplete cellular ATP, and SNF1 is essential in the response to glucose starvation. In both cases, activation requires phosphorylation at a conserved threonine residue within the activation loop of the kinase domain, but identifying the upstream kinase(s) responsible for this has been a challenging, unsolved problem. RESULTS: Using a library of strains that express 119 yeast protein kinases as GST fusions, we identified Elm1p as the sole kinase that could activate the kinase domain of AMP-activated protein kinase in vitro. Elm1p also activated the purified SNF1 complex, and this correlated with phosphorylation of Thr210 in the activation loop. Removal of the C-terminal domain increased the Elm1p kinase activity, indicating that it is auto-inhibitory. Expression of activated, truncated Elm1p from its own promoter gave a constitutive pseudohyphal growth phenotype that was rescued by deletion of SNF1, showing that Snf1p was acting downstream of Elm1p. Deletion of ELM1 does not give an snf- phenotype. However, Elm1p is closely related to Pak1p and Tos3p, and a pak1Delta tos3Delta elm1Delta triple mutant had an snf1- phenotype, i.e., it would not grow on raffinose and did not display hyperphosphorylation of the SNF1 target, Mig1p, in response to glucose starvation. CONCLUSIONS: Elm1p, Pak1p, and Tos3p are upstream kinases for the SNF1 complex that have partially redundant functions.

AMP-Activated Protein Kinase Kinases↗
PubMed2003

Improvement of optical resolution in far-field imaging by optical multiplication.

Based on the idea of the complex filters suggested by Toraldo di Francia in 1952, super-resolved imaging has been achieved by using interferometric image multiplication. The question of whether super-resolution implies a limited field of view is discussed, and an analytical criterion to decide this question is obtained. Three different imaging configurations are investigated, all of which gave a spatial resolution exceeding that corresponding to the full open aperture of the optical system. A resolution limit 55% of the Rayleigh limit was achieved for incoherently illuminated objects. One price that must be paid for the super-resolution is inefficient use of the light source. A second price is a limitation to the usable field of view.

Fourier Analysis↗
PubMed2003

The relationship between changes in intrinsic optical signals and cell swelling in rat spinal cord slices.

Changes in intrinsic optical signals could be related to cell swelling; however, the evidence is not compelling. We measured light transmittance, ECS volume fraction (alpha), and extracellular K+ in rat spinal cord slices during electrical stimulation and the application of elevated potassium, NMDA, or anisoosmotic solutions. Dorsal root stimulation (10 Hz/1 min) induced an elevation in extracellular K+ to 6-8 mM, a light transmittance increase of 6-8%, and a relative ECS volume decrease of less than 5%; all of these changes had different time courses. The application of 6 or 10 mM K+ or NMDA (10(-5) M) had no measurable effect on alpha, but light transmittance increased by 20-25%. The application of 50 or 80 mM K+ evoked a 72% decrease in alpha while the light transmittance increase remained as large as that in 6 or 10 mM K+. While the change in alpha persisted throughout the 45-min application, light transmittance, after peaking in 6-8 min, quickly returned to control levels and decreased below them. Astrocytic hypertrophy was observed in 6, 10, and 50 mM K+. The same results followed the application of 10(-4) M NMDA or hypotonic solution (160 mmol/kg). The elevation of extracellular K+ after NMDA application, corresponding to increased neuronal activity, had a similar time course as the light transmittance changes. Furosemide, Cl(-)-free, or Ca(2+)-free solution blocked or slowed down the decreases in alpha, while the light transmittance increases were unaffected. In hypertonic solution (400 mmol/kg), alpha increased by 30-40%, while light transmittance decreased by 15-20%. Thus, light transmittance changes do not correlate with changes in ECS volume but are associated with neuronal activity and morphological changes in astrocytes.

Animals↗
PubMed2002

Mouse resident peritoneal macrophages partially control in vitro infection with Coxiella burnetii phase II.

Coxiella burnetii, the agent of Q fever in man and of coxiellosis in other species, is a small, dimorphic, obligate intracellular bacterium, sheltered within large, acidified, and hydrolase-rich phagosomes. Although several primary and established cell lines, macrophage-like cells, and primary macrophages from other species have been infected with C. burnetii, the infection of mouse primary macrophages has not been sufficiently characterized. In this report quantification of DAPI (4', 6-diamino-2-phenylindole) fluorescence images acquired by confocal microscopy, and transmission electron microscopy were used to compare the infection of three mouse-derived cells, L929 fibroblasts, J774 macrophage-like cells, and resident peritoneal macrophages, with a phase II clone of C. burnetii known to be non-virulent for mammals. Infected peritoneal phagocytes differed from L929 or J774 cells in that: (a) large vacuoles took longer to appear (3-5 d instead of 2), and were only found in a subset (20-30%) of macrophages, as opposed to in more than 70% of the other cells; (b) total and vacuole-associated relative bacterial loads in L929 and J774 cells were several-fold higher than in peritoneal macrophages; (c) estimated doubling times of the bacteria were about 68 h in the primary macrophages, 18 h in J774 and 22 h in L929 cells. Thus, mouse resident peritoneal macrophages control both the formation of the large vacuoles and the intracellular proliferation of C. burnetii phase II.

Animals↗
PubMed2001

Massaliatrema misgurni n. sp. (Trematoda: Heterophyidae) whose metacercariae encyst in loaches (Misgurnus anguillicaudatus).

Hitherto unknown metacercariae were found encysted in loaches (Misgurnus anguillicaudatus) from China. They were experimentally fed to golden hamsters, and gravid adults were recovered 1 week post-infection from their small intestines. A new species, Massaliatrema misgurni n. sp. (Heterophyidae), is described from the adults. This new species is different from M. gyrinicola Dollfus and Timon-David, 1960, in having a smaller acetabulum/oral sucker ratio, less branched vitellaria widely entering the intercecal anteroacetabular area and an almost median seminal receptacle; and from M. yamashitai Kamiya and Ohbayashi, 1975, in having a larger acetabulum/oral sucker ratio, with the seminal vesicle situated in the uterine loop and the vitellaria entering the intercecal anteroacetabular area. This report is the first record of M. anguillicaudatus as a second intermediate host of the genus Massaliatrema Dollfus and Timon-David, 1960.

Animals↗
PubMed2002

Do Plasmodium ookinetes invade a specific cell type in the mosquito midgut?

Recent debate in Plasmodium ookinete invasion has been centered on whether the parasite chooses a specific cell type to cross the midgut epithelium in the mosquito. A few publications have described the mosquito midgut being composed of complex surface-structures, histochemically and biochemically diverse cell types, and have proposed that Plasmodium gallinaceum ookinetes prefers a specific cell type (Ross cell) in Aedes aegypti for crossing the midgut epithelium. Two recent publications reported, however, that with differential interference contrast microscopy, all midgut epithelial cells in uninfected mosquitoes appear structurally similar and argued that ookinetes do not invade a specific cell type. These observations are discussed here in the context of the 'Ross cell' hypothesis.

Aedes↗
PubMed2006

High-magnification ICSI overcomes paternal effect resistant to conventional ICSI.

Previous studies have shown that repeated intracytoplasmic sperm injection (ICSI) failures can be caused by a paternal effect. Other studies have suggested that ICSI results are compromised if morphologically abnormal spermatozoa are injected into oocytes. This study was undertaken to evaluate the usefulness of a high-magnification optical system to select spermatozoa to be used for ICSI (high-magnification ICSI) in couples with repeated conventional ICSI failures. Couples with two or more previous conventional ICSI failures underwent an additional conventional ICSI attempt, followed by a high-magnification ICSI attempt. The outcomes of the two sequential attempts were compared. In 72 of these patients, sperm DNA integrity was assessed. In the whole group of 125 couples with repeated ICSI failures, high-magnification ICSI improved clinical outcomes (pregnancy, implantation, delivery and birth rates) without affecting biological outcomes (fertilization and cleavage rates, embryo morphology). The improvement of clinical ICSI outcomes was evident both in patients with an elevated degree of sperm DNA fragmentation and in those with normal sperm DNA status. It is concluded that high-magnification ICSI improves clinical outcomes in couples with previous repeated conventional ICSI failures.

Adult↗
PubMed1975

Variations in the water content of the larvae of Nematodirus battus during the hatching process.

During a series of experiments on the ecology of Nematodirus battus observations were made on the uptake of water by eggs containing third-stage larvae. It was found that, as they approached hatching, the water content of the larvae increased markedly, then fell immediately after they escaped from the shell. The method of measuring the water content, using interferometry, is also described.

Animals↗
PubMed1981

The initial stages of infection of cattle cells with Theileria parva sporozoites in vitro.

Large numbers of Theileria parva sporozoites were separated from Rhipicephalus appendiculatus adult ticks by filtration and were then concentrated by centrifugation. The sporozoites were incubated at 37 degrees C with leucocytes from 6 cattle of Bos indicus and B. taurus types. Giemsa-stained smears and living preparations under interference contrast microscopy were used to follow the course of the infection of the leucocytes with sporozoites. Sporozoites were seen to attach rapidly to about 25% of the leucocytes which they penetrated. After penetration by the sporozoites the morphology of the cells changed to show an increase in cytoplasm and an enlargement of the Golgi apparatus, with which the parasite appeared to become associated. The early intracellular or preschizont stages resembled Babesia parasites. From day 3, the parasite showed the typical morphology of the macroschizont of T. parva. Multiple infections were frequent and up to 8 schizonts were observed arranged around the Golgi apparatus. Multiple infected cells did not survive in culture but some of the cells infected with a single parasite divided to produce 2 infected daughter cells and infected lymphoblastoid cell lines were established in all 21 attempts.

Animals↗