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Biomedical subjects

D Ritter

Publications and source records attributed to D Ritter.

At least 37 records · Page 2Linked to original sources

Differentiation of Mycobacterium genavense and Mycobacterium simiae by automated mycolic acid analysis with high-performance liquid chromatography.

Mycobacterium genavense, a fastidious opportunist in patients with AIDS, cannot be identified by conventional biochemical methods. Computerized mycolic acid analysis by high-performance liquid chromatography offers an alternative that distinguishes the mycolic acid profile of M. genavense from those of all other organisms in the database developed at the Centers for Disease Control and Prevention.

Bacterial Typing Techniques↗

Distribution and regulation of guanylyl cyclase type B in the rat nephron.

C-type natriuretic peptide (CNP) has been localized to the proximal and distal nephron. In this study, we examined the distribution and regulation of the CNP receptor, guanylyl cyclase type B (GC-B), in the rat kidney. GC-B mRNA was detected most frequently in microdissected glomeruli, thin and thick limbs of the loop of Henle, and outer and inner medullary collecting ducts by reverse transcription-polymerase chain reaction (RT-PCR). This pattern of expression is supported by immunofluorescent staining, using anti-GC-B-specific antiserum. Nearly equivalent levels of GC-B and guanylyl cyclase type A (GC-A) mRNAs were found by quantitative RT-PCR (5,662 +/- 1,622 and 5,187 +/- 1,204 molecules of cDNA/microgram total RNA, respectively; means +/- SE, n = 6). Renal inner medulla GC-B mRNA levels, but not renal CNP mRNA levels, were 3.2-fold greater in hypervolemic and 2.3-fold less in hypovolemic rats compared with euvolemic controls. Immunohistochemical staining also supports a greater GC-B expression with increased volume status. These data link hydration status and GC-B expression and suggest an additional and novel mechanism for regulating intravascular volume.

Animals↗

Axial muscle function during lizard locomotion

It was recently reported that the epaxial muscles of a lizard, Varanus salvator, function to stabilize the trunk during locomotion, and it was suggested that this stabilizing role may be a shared derived feature of amniotes. This result was unexpected because it had previously been assumed that the epaxial muscles of lizards function to produce lateral bending during locomotion and that only in mammals and birds were the epaxial muscles active in stabilizing the trunk. These results and the inferences made from them lead to two questions. (1) Is the pattern of epaxial muscle activity observed in V. salvator representative of a basal lizard condition or is it a derived condition that evolved within lizards? (2) If the epaxial muscles do not produce lateral bending, which muscles do carry out this function? These questions were addressed by collecting synchronous electromyographic (EMG) and kinematic data from two lizard species during walking and running. EMG data were collected from the epaxial muscles of a lizard species from a basal clade, Iguana iguana, in order to address the first question. EMG data were collected from the hypaxial muscles of both Iguana iguana and Varanus salvator to address the second question. The timing of epaxial muscle activity in Iguana iguana relative to the kinematics of limb support and lateral trunk bending is similar to that observed in Varanus salvator, a finding that supports the hypothesis that the epaxial muscles stabilize the trunk during locomotion in lizards and that this stabilizing role is a basal feature of lizards. Therefore, a stabilizing function of the epaxial muscles is most parsimoniously interpreted as a basal amniote feature. In both Iguana iguana and Varanus salvator, the activity of two of the hypaxial muscles, the external oblique and rectus abdominis, is appropriately timed for the production of lateral bending. This indicates that elements of the hypaxial musculature, not the epaxial musculature, are the primary lateral bending muscles of lizards.

Journal Article↗

Natriuretic peptide receptors A and B have different cellular distributions in rat kidney.

The cellular distribution of guanylyl cyclase coupled natriuretic peptide receptors type A (GC-A) and type B (GC-B) was examined by immunocytochemistry in normal rat kidney, and compared with the distribution of the vacuolar H(+)-ATPase. Staining for GC-A was found in glomeruli, thin limbs of Henle's loop, cortical collecting tubule, and inner medullary collecting duct. Staining for GC-B was found in glomeruli and the same nephron sections as GC-A, with the exception of the thin limbs. In the cortical collecting tubule, GC-A was found in both principal and intercalated cells; GC-B was restricted to the apical pole of alpha intercalated cells. In inner medullary collecting duct cells, GC-A was located on the basal membrane, whereas GC-B was found in the apical pole. The different pattern of polarization of natriuretic peptide receptors in the inner medulla provides a plausible basis for the different physiologic effects of atrial natriuretic factor and C-type natriuretic peptide. The results also suggest the possibility that GC-B is involved in the regulation of bicarbonate transport in the cortical collecting tubule.

Animals↗

Polarized distribution of renal natriuretic peptide receptors in normal physiology and ischemia.

The polarized expression of guanylyl cyclase-coupled natriuretic peptide receptors, types A (GC-A) and B (GC-B), was measured in inner medullary collecting ducts (IMCD) of normal and ischemic rat kidneys, as well as in IMCD cells. Exposure of normal rat kidney medulla to an anti-GC-A antibody demonstrated a propensity of receptor staining on the cellular basal membrane. The polarization of GC-A receptors was lost in the ischemic kidney. The maximal binding capacity of 125I-atrial natriuretic factor (ANF) to the basal membrane of the inner medullary cell line mIMCD-K2 was five times greater than that to the apical membrane. ANF or C-type natriuretic peptide (CNP) added to the basal side of cultured cells resulted in guanosine 3',5'-cyclic monophosphate formation that was greater than when applied to the apical side. Depletion of ATP stores in cultured cells was followed by an increase of 125I-ANF binding to apical cellular membranes. Similar results were obtained when receptor guanylyl cyclase activity was assayed. In conclusion, these results suggest that functional GC-A and GC-B receptors are present predominantly on the basal membrane of IMCD. However, depletion of cellular ATP stores such as in ischemia is followed by a partial loss of polarization.

Animals↗

Epaxial muscle function during locomotion in a lizard (Varanus salvator) and the proposal of a key innovation in the vertebrate axial musculoskeletal system

The pattern of electromyographic activity in the epaxial muscles of walking and running lizards (water monitors, Varanus salvator) was quantified with high-speed video and synchronized electromyography. Muscle denervation experiments were performed and ground reaction forces were recorded to evaluate hypotheses of muscle function. Water monitors exhibit unilateral, uniphasic activation of the epaxial muscles ipsilateral to rear limb support. The iliocostalis and longissimus dorsi muscles are active throughout the support phase of the rear limb, while the transversospinalis is active only during the second half of the support phase. The timing of muscle activity does not support the presumed locomotor function of the epaxial muscles of lizards (lateral bending of the trunk). Bilateral denervation of the two most lateral epaxial muscles (iliocostalis and longissimus dorsi) caused no significant change in the amplitude of lateral trunk bending. These two epaxial muscles do not appear to be necessary for lateral bending of the trunk, nor do they appear to be involved in resisting lateral bending of the trunk. On the basis of the data reported in this study, as well as a re-evaluation of previously existing data, it is proposed that the locomotor function of the epaxial muscles of lizards is to stabilize the trunk against the vertical component of the ground reaction forces; i.e. the epaxial muscles function in a postural role during locomotion in lizards, as they do in other amniotes. This proposed postural role of the epaxial muscles may represent a key innovation of amniotes, made possible by the loss of an aquatic larval stage in their common ancestor.

Journal Article↗

The kinematics and mechanism of prey capture in the African pig-nosed frog (Hemisus marmoratum): description of a radically divergent anuran tongue.

High-speed videography and muscle denervation experiments were used to quantify the feeding kinematics of Hemisus marmoratum and to test hypotheses of muscle function. The feeding behavior of H. marmoratum, which feeds on ants and termites, differs radically from that of other frogs that have been studied. During feeding in H. marmoratum, the tongue 'telescopes' straight out of the mouth, as opposed to the 'flipping' tongue trajectory observed in most other frogs. At the time of prey contact, two lateral lobes of tissue at the tongue tip envelop the prey. These lateral lobes are capable of applying significant pulling forces to the prey and the tongue is, therefore, described as prehensile. The trajectory of the tongue can be adjusted throughout protraction so that the frog can 'aim' its tongue in all three dimensions; distance, azimuth and elevation. Bilateral denervation of the genioglossus muscles results in a complete lack of tongue protraction, indicating that the genioglossus muscle is the main tongue protractor in H. marmoratum, as in other frogs. Thus, H. marmoratum provides strong evidence of functional conservatism of the genioglossus muscle within anurans. Bilateral denervation of the hyoglossus muscle indicates that although the hyoglossus is involved in several aspects of normal tongue retraction, including the prehensile capability of the tongue tip, it is not necessary for tongue retraction. Unilateral denervation of the genioglossus muscle causes significant deviation of the tongue towards the denervated side, providing evidence for a mechanism of lateral tongue aiming. On the basis of the kinematics of prey capture, the anatomy of the tongue and the results of the denervation experiments, we propose that H. marmoratum uses a hydraulic mechanism to protract its tongue.

Animals↗

Endogenous serum antibodies that interfere with a common thyroid hormone uptake assay: characterization and prevalence.

We identified individuals whose serum contained a substance that produced falsely decreased thyroid hormone (T)-uptake values determined by the Emit (Syva) procedure. Investigation of this interference was prompted by identification of a patient with T-uptake values inconsistent with clinical assessment. IgG depletion and supplementation studies with this patient's serum suggested that the interference was due to endogenous antibodies with specificity for the thyroxine-glucose-6-phosphate dehydrogenase conjugate in the Emit T-uptake assay. The prevalence of the interference was examined by prospectively comparing routine Emit T-uptake values of 1710 patients' samples to T-uptake values obtained by another method. Discrepant samples were also assayed by a radioactive binding triiodothyronine-uptake assay. We identified eight samples that had falsely decreased T-uptake values by Emit, for an overall prevalence of 0.46%. Among 45 consecutive patients with a T-uptake value < 20%, five patients, or 11%, were falsely decreased by Emit and three of these were clearly due to an interfering IgG. We suggest that samples with abnormally low T-uptake values determined by the Emit method be confirmed by an alternative method.

Adult↗

Seroprevalence of hepatitis B viral markers in 52,000 Alaska Natives.

As a part of a program to screen and immunize as many Alaska Natives as possible against hepatitis B infection, hepatitis B virus seromarkers were measured in 52,022 Alaska Natives between 1983 and 1987. Hepatitis B surface antigen (HBsAg) was found in 1,603 persons (3.1%), and 7,155 persons (13.8%) exhibited hepatitis B virus seropositivity. While the prevalence of total seropositivity increased with increasing age (p < 0.001), the proportion of seropositive persons who were also positive for HBsAg was significantly higher in children under age 5 years than in persons over age 60 (p < 0.001). The total hepatitis B virus seropositivity was significantly higher in males than in females (p < 0.001), and a greater proportion of seropositive males than seropositive females had HBsAg (p < 0.001). The prevalence of HBsAg and total seropositivity differed significantly by geographic region (p < 0.001), ranging from 0.5% to 8.2% for presence of HBsAg and from 5.4% to 29% for total seropositivity. Of persons who were HBsAg positive, hepatitis B e antigen was found in 35.4%, and antibody to hepatitis e antigen was found in 49.6%. The prevalence of hepatitis B e antigen significantly decreased with age, while that of antibody to hepatitis e antigen significantly increased. The prevalence of hepatitis B virus seropositivity is high in Alaska Natives, and there are significant differences in both the prevalence of HBsAg and total hepatitis B virus seromarkers by age, sex, and geographic region.

Adolescent↗

Positron emission tomography in myotonic dystrophy.

Regional cerebral glucose metabolism (rCMRG1) was studied in three patients with myotonic dystrophy by positron emission tomography with 18F-2-fluoro-2-deoxy-D-glucose as the radiotracer. Clinically, all patients presented an organic personality syndrome. Impairment of rCMRG1 was found in all cortical and subcortical regions, particularly in the frontal cortex and the lentiform nucleus. The results suggest that the organic personality changes that occur in myotonic dystrophy reflect widespread cerebral pathology. The metabolic pattern was similar to that seen in normal aging but was qualitatively more severe and is therefore consistent with the classification of myotonic dystrophy as a progeric disease.

Adult↗

Endogenous antibodies that interfere with thyroxine fluorescence polarization assay but not with radioimmunoassay or EMIT.

We have identified an individual whose thyroxine (T4) concentration was undetectable with Abbott's fluorescence polarization immunoassay (FPIA) but within the reference range by radioimmunoassay or EMIT (Syva). The patient's thyrotropin, triiodothyronine, and T-uptake values were within the normal range. The T4 concentration measured by FPIA increased to normal when the immunoglobulin fraction was selectively removed from the serum. When the patient's immunoglobulin fraction was added to normal serum, the T4 content of the normal serum measured by FPIA became falsely low. The patient's antibody interfered with the T4 FPIA by binding to the fluorescein-T4 conjugate. The T-uptake was less affected by the patient's serum because of the low affinity of the patient's antibody to fluorescein-T4 (K = 3.5 x 10(8) L/mol). The patient's immunoglobulin bound preferentially to fluorescein-T4, in comparison with binding to fluorescein or T4 alone. We conclude that the patient's immunoglobulin bound to an epitope unique to the fluorescein-conjugated T4.

Adult↗

Processing of atriopeptin prohormone by nonmyocytic atrial cells.

Atriopeptin (AP) is synthesized and stored in the mammalian atria as a 126 amino acid prohormone (AP126). Upon secretion, the prohormone undergoes site specific proteolysis within the atria to yield the carboxyl terminal 28 amino acid hormone (AP28). The atrial cell responsible for AP126 bioactivation has not yet been determined. Primary neonatal rat atrial cell cultures were generated with and without depletion of nonmyocytic cells. The molecular form of AP detected in the conditioned media of mixed cultures was determined to be AP126. Addition of dexamethasone to these cultures resulted in the appearance of a peptide that co-migrated with AP28. In contrast, no AP126 processing was detected in the conditioned media of myocyte enriched cultures when grown in the presence of dexamethasone. Readdition of nonmyocytic atrial cells to myocyte enriched cultures successfully reconstituted the steroid induced AP126 processing. Incubation of recombinant AP126argarg with nonmyocytic atrial cell cultures resulted in the generation of AP28argarg. We conclude that a nonmyocytic atrial cell is responsible for AP126 processing in vitro.

Animals↗

Tumour necrosis factor alpha restores granulomas and induces parasite egg-laying in schistosome-infected SCID mice.

Schistosomiasis (bilharzia) is a parasitic disease caused by several species of schistosome worms (blood flukes). The key pathogenic event in this disease is the formation of granulomas around schistosome eggs trapped in portal venules of the liver. Granulomas are a distinctive form of chronic inflammation characterized by localized aggregation of activated macrophages around an inciting stimulus. Each granuloma evolves to form a fibrous scar; in schistosomiasis, the result is widespread hepatic fibrosis and portal hypertension. To identify the specific immune signal molecules necessary for granuloma formation, we studied schistosome infections in severe combined immunodeficient (SCID) mice, which have normal macrophages but lack functional B or T lymphocytes. Here we report that the immunoregulatory cytokine tumour necrosis factor alpha is necessary and sufficient to reconstitute granuloma formation in schistosome-infected SCID mice. Moreover, we find that the parasitic worms require tumour necrosis factor alpha for egg-laying and for excretion of eggs from the host. The implication of this latter result is that the parasite has adapted so successfully to its host that it uses a host-derived immunoregulatory protein as a signal for replication and transmission.

Animals↗

Response to hepatitis B vaccine of persons positive for antibody to hepatitis B core antigen.

The significance of antibody to hepatitis B core antigen (anti-HBc) present in a person's serum without hepatitis B surface antigen (HBsAg) or its antibody (anti-HBs) is unknown. Serum specimens from 281 persons initially positive only for anti-HBc by enzyme immunoassay (EIA) were retested by radioimmunoassay (RIA), and of these, 177 (63%) remained positive for anti-HBc by both assays. Of these 177 persons, 3 were positive for HBsAg, and 72 possessed low levels of anti-HBs [less than 10 sample ratio units; (SRU's)]. When persons positive for anti-HBc by EIA and RIA were given one 20-micrograms dose of plasma-derived hepatitis B vaccine and tested for anti-HBs 1 month later, a booster response was observed in 14 of 41 (34%) persons with low level anti-HBs and 3 of 50 (6%) persons negative for anti-HBs. Of those positive only for anti-HBc by EIA but negative by RIA, only 3 of 37 (8.1%) showed a booster response. Of those who completed the three-dose immunization series and did not show a booster response, 63 of 80 (78.8%) developed anti-HBs levels greater than 10 standard ratio unit. The majority of persons with isolated anti-HBc will have a primary rather than a booster response to hepatitis B vaccine.

Adolescent↗

Localization, synthetic regulation, and biology of renal atriopeptin-like prohormone.

We recently demonstrated the synthesis and secretion of an atriopeptin (AP)-like prohormone in rat neonatal and adult cortical kidney cell cultures. However, these cultures contained proximal as well as distal tubular epithelial cells; thus characterization of the peptide synthetic cell was not possible. Also, by immunohistochemical techniques, we localized this AP-like prohormone to the distal cortical nephron in adult rat kidney. In this study, we examined further details of the kidney cortical cell type that expresses and secretes this AP-like peptide in adult renal cortical cell cultures, its regulation by adenylate cyclase via adenosine 3',5'-cyclic monophosphate (cAMP) generation, and its ability to stimulate guanylate cyclase. Tubular fragments were derived from cortical tissue of adult Sprague-Dawley rats and separated into four fractions on Percoll density gradient. Cell cultures generated from fraction 3 secreted 5- to 10-fold the amount of this renal peptide compared with fractions 2 and 4. Further cell culture characterization was performed by agonist-stimulated cAMP formation, kallikrein localization, and prostaglandin E2 formation. From these analyses, it was determined that tissue band 3 was enriched for distal cortical connecting tubules. To further evaluate whether mammalian distal nephron synthesizes an AP-like protein, we determined that two immortalized mouse cell lines, derived from either the distal convoluted tubule or cortical collecting tubule, synthesized a radiolabeled AP after being pulsed with [35S]-methionine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗