Biomedical subjects

J M Kramer

Publications and source records attributed to J M Kramer.

At least 19 recordsLinked to original sources

PubMed2001

Chronic exercise increases GAD gene expression in the caudal hypothalamus of spontaneously hypertensive rats.

Previous studies have suggested that a gamma-amino-butyric acid (GABA) deficit in the caudal hypothalamus (CH) of the spontaneously hypertensive rat (SHR) contributes to elevated levels of arterial pressure. The purpose of this study was to examine if SHR that underwent exercise training demonstrated a blunted development of hypertension and greater levels of glutamic acid decarboxylase (GAD) mRNA transcripts in the caudal hypothalamus. SHR were randomly paired and assigned to either a trained group (T; n=9) or a non-trained control group (NT; n=9). Trained animals were exercised for 10 weeks on a motorized treadmill while NT animals concurrently rested on a mock-treadmill. Following the 10-week training period, Northern blot analyses of mRNA for both the 65-kDa (GAD(65)) and 67-kDa (GAD(67)) isoforms of GAD were performed on tissue from caudal hypothalamic and cerebellar control brain regions. Exercise training simultaneously blunted the developmental rise in blood pressure in SHR (Delta59+/-9 mmHg in trained versus Delta77+/-9 mmHg in non-trained; P<0.03) and increased both GAD(65) (147+/-44%) and GAD(67) (162+/-77%) mRNA transcript levels in the CH (P<0.05). In contrast, no difference was detected in GAD mRNA levels in the cerebellum between T and NT SHR. These findings are consistent with our previous functional studies and demonstrate that exercise can significantly and specifically upregulate GAD gene transcript levels in the caudal hypothalamus of hypertensive rats.

Animals↗

PubMed2001

The NC1/endostatin domain of Caenorhabditis elegans type XVIII collagen affects cell migration and axon guidance.

Type XVIII collagen is a homotrimeric basement membrane molecule of unknown function, whose COOH-terminal NC1 domain contains endostatin (ES), a potent antiangiogenic agent. The Caenorhabditis elegans collagen XVIII homologue, cle-1, encodes three developmentally regulated protein isoforms expressed predominantly in neurons. The CLE-1 protein is found in low amounts in all basement membranes but accumulates at high levels in the nervous system. Deletion of the cle-1 NC1 domain results in viable fertile animals that display multiple cell migration and axon guidance defects. Particular defects can be rescued by ectopic expression of the NC1 domain, which is shown to be capable of forming trimers. In contrast, expression of monomeric ES does not rescue but dominantly causes cell and axon migration defects that phenocopy the NC1 deletion, suggesting that ES inhibits the promigratory activity of the NC1 domain. These results indicate that the cle-1 NC1/ES domain regulates cell and axon migrations in C. elegans.

Amino Acid Sequence↗

PubMed2001

Coinfection with Campylobacter species: an epidemiological problem?

AIMS: To determine the frequency of coinfection with multiple strains in sporadic cases of human Campylobacter infection. METHOD AND RESULTS: During 1999 10 single colonies of Campylobacter were cultured from each of 53 positive faecal samples. Five isolates were taken from nonselective agar after passive filtration of faecal suspensions and five isolates were taken from selective agar plates. All isolates were sero- and phage typed and their antibiotic resistance determined. Pulsed-field gel electrophoresis and flagellin gene typing were performed on selected isolates. One patient was infected with Camp. coli, the remainder with strains of Camp. jejuni. The majority of patients was infected with a single strain of Campylobacter, but from each of four samples, 7.5%, two strains of Camp. jejuni, confirmed by molecular typing, were identified. CONCLUSION: Coinfection occurs in sporadic cases of campylobacteriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has implications in outbreak investigation when distinct strains have been isolated from epidemiologically related patients and/or the suspected source or vehicle.

Adult↗

PubMed2001

Spontaneously hypertensive rats exhibit altered cardiovascular and neuronal responses to muscle contraction.

We examined the cardiovascular and ventrolateral medullary neuronal responses to muscle contraction in the spontaneously hypertensive rat (SHR) and normotensive Wistar-Kyoto rat (WKY) control. Cardiovascular, respiratory and ventrolateral medullary neuronal responses to muscle contraction evoked by tibial nerve stimulation were recorded. SHRs exhibited significantly larger drops in arterial pressure compared to WKYs in response to muscle contraction (P < 0.05). Basal ventrolateral medulla neuronal discharge rates were similar between the SHR and the WKY groups. A majority of neurons recorded responded to muscle contraction in both the WKY (77 %; n = 53) and the SHR groups (68 %; n = 62). There was no difference in the percentage of neurons that responded with an increase (approximately 60 %) or decrease (approximately 40 %) in firing rate between hypertensive and normotensive rats. Pulse wave-triggered averaging techniques showed that most neurons that responded to muscle contraction also possessed a basal firing rhythm temporally related to the cardiac cycle (85 % in WKYs, 83 % in SHRs). However, decreases in neuronal firing rates in response to muscle contraction were significantly greater in SHRs than WKYs. Therefore, we conclude that muscle contraction unmasks a hyperexcitability of neurons in the ventrolateral medulla of SHRs that parallels the heightened blood pressure responses.

Animals↗

PubMed2001

Chronic exercise alters caudal hypothalamic regulation of the cardiovascular system in hypertensive rats.

Previous studies have documented a deficit in the GABA neurotransmitter system within the caudal hypothalamus (CH) of spontaneously hypertensive rats (SHR). The reduction in inhibitory influence on this cardiovascular excitatory brain region is associated with an increased neuronal activity and resting blood pressure. The purpose of this study was to determine if chronic treadmill and wheel-running activities alter the ability of the CH to regulate cardiovascular function. SHR were exercised on a treadmill (5 times/wk) at moderate intensity or allowed free access to running wheels (7 days/wk) for a period of 10 wk. Resting blood pressures were obtained before and after the exercise training periods. After the exercise period, rats were anesthetized and microinjection experiments were performed. Treadmill-trained SHR exhibited a significantly blunted developmental rise in resting blood pressure after 10 wk of exercise. A similar yet less marked effect was observed in wheel-run rats. Microinjection of the GABA synthesis inhibitor 3-mercaptopropionic acid (3-MP) into the CH of nonexercised SHR did not produce any change in arterial pressure. In contrast, microinjection of 3-MP into the CH produced significant increases in blood pressure and heart rate in exercised SHR. These results demonstrate that exercise training can alter CH cardiovascular regulation in hypertensive rats and therefore may play a role in increasing cardiovascular health.

3-Mercaptopropionic Acid↗

PubMed2001

Reflex cardiovascular responses originating in exercising muscles of mice.

The cardiovascular responses induced by exercise are initiated by two primary mechanisms: central command and reflexes originating in exercising muscles. Although our understanding of cardiovascular responses to exercise in mice is progressing, a murine model of cardiovascular responses to muscle contraction has not been developed. Therefore, the purpose of this study was to characterize the cardiovascular responses to muscular contraction in anesthetized mice. The results of this study indicate that mice demonstrate significant increases in blood pressure (13.8 +/- 1.9 mmHg) and heart rate (33.5 +/- 11.9 beats/min) to muscle contraction in a contraction-intensity-dependent manner. Mice also demonstrate 23.1 +/- 3.5, 20.9 +/- 4.0, 21.7 +/- 2.6, and 25.8 +/- 3.0 mmHg increases in blood pressure to direct stimulation of tibial, peroneal, sural, and sciatic hindlimb somatic nerves, respectively. Systemic hypoxia (10% O(2)-90% N(2)) elicits increases in blood pressure (11.7 +/- 2.6 mmHg) and heart rate (42.7 +/- 13.9 beats/min), while increasing arterial pressure with phenylephrine decreases heart rate in a dose-dependent manner. The results from this study demonstrate the feasibility of using mice to study neural regulation of cardiovascular function during a variety of autonomic stimuli, including exercise-related drives such as muscle contraction.

Animals↗

PubMed2001

Disseminated vaccine strain varicella as the acquired immunodeficiency syndrome-defining illness in a previously undiagnosed child.

The Food and Drug Administration licensed a live-virus varicella vaccine (Varivax; Merck & Co Inc, West Point, PA) in March 1995. Prelicensure adverse events were minimal; however, since licensure and increased vaccine use, rare previously undetected risks have arisen. Presented here is the clinical course of a previously undiagnosed, human immunodeficiency virus-infected boy who developed dissemination of the vaccine strain of varicella zoster after immunization. chickenpox, human immunodeficiency virus, pneumonia, encephalopathy, varicella vaccine, adverse events, dissemination.

Chickenpox↗

PubMed2001

Cardiopulmonary function in rats with lung hemorrhage induced by pulsed ultrasound exposure.

OBJECTIVE: To assess cardiopulmonary function in rats exposed to pulsed ultrasound using superthreshold exposure conditions known to produce significant lung hemorrhage. METHODS: In 1 group of 9 anesthetized Sprague-Dawley rats, 5 foci of ultrasound-induced hemorrhage were produced in the left lung of each rat. In a second group of 6 rats, 5 foci of ultrasound-induced hemorrhage were produced in the left and right lungs of each rat. Each lesion was induced using superthreshold pulsed ultrasound exposure conditions (3.1-MHz center frequency, 1.7-kHz pulse repetition frequency, 1.3-micro-second pulse duration, 60-second exposure duration, 39-MPa in situ peak compressional pressure, and 17-MPa in situ peak rarefactional pressure). After exposure, the lungs were fixed in formalin and assessed histologically. The total lesion volume was calculated for each lesion in each lung lobe. Measurements of cardiopulmonary function included assessment of pulsatile arterial pressure, heart rate, end-tidal carbon dioxide, respiratory rate, and arterial blood gases (PCO2 and PO2). Functional data were quantified before (baseline) and 30 minutes after exposure to ultrasound. RESULTS: In the 9 rats that had lesions in only the left lung, the mean (SEM) lesion volume was 97 (13) mm3 and represented about 3.4% of the total lung volume. In the 6 rats that had lesions in both the left and right lungs, the left, right, and total mean lesion volumes, respectively, were 102 (16), 114 (11), and 216 (18) mm3 and represented about 3.7%, 4.2%, and 7.9% of the total lung volume. There were no statistically significant differences in cardiopulmonary measurements between baseline values and values obtained after exposure to ultrasound in the 9 rats exposed on the left lung only. The 6 rats exposed bilaterally had statistically significant differences in arterial pressure (134 +/- 4 versus 113 +/- 9 mm Hg; P= .047) and arterial PO2 (70 +/- 5 versus 58 +/- 4 mm Hg; P = .024) between baseline values and values obtained after exposure to ultrasound. CONCLUSIONS: The severity of ultrasound-induced lesions produced in 1 lung did not affect measurements of cardiopulmonary function because of the functional respiratory reserve in the unexposed lung. However, when both the left and right lungs had ultrasound-induced lesions, the functional respiratory reserve was decreased to a point at which rats were unable to maintain systemic arterial pressure or resting levels of arterial PO2.

Animals↗

PubMed2001

The palm as a real-time wide-area data-access device.

Handheld wireless technologies offer great promise in helping to improve healthcare. However, it is not clear whether off-the-shelf wireless networking will work as well within medical centers as this technology works outside of the medical center. Therefore, we evaluated the coverage of wide-area wireless technology within two representative academic medical centers. The study determined the rate of connectivity by testing both the Palm VII and the Minstrel V modem in a set of locations typically frequented by house staff in their daily activities. Within one hospital, connectivity was 59% for OmniSky service, and 78% for Palm.net. The second hospital's connectivity was over 93% with both devices. Differences in connectivity were likely due to the number of rooms visited with externally exposed walls, the suburban versus urban location of the academic medical center, and the relative location of service transponders. When examined by the Johns Hopkins Clinical Devices Laboratory, both devices were found to operate without interfering with other hospital equipment.

Academic Medical Centers↗

PubMed2000

Hypothalamus, hypertension, and exercise.

The hypothalamus is a well-known autonomic regulatory region of the brain involved in integrating several behaviors as well as cardiorespiratory activity. Our laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses associated with exercise. In addition, other findings from this laboratory and others have implicated alterations in this same brain region in spontaneously hypertensive rats as contributing factors of the elevated levels of arterial pressure in hypertension. Several studies have revealed a gamma-amino-butyric acid (GABAergic) deficiency in the caudal hypothalamus of spontaneously hypertensive rats that contributes to the tonic disinhibition and overactivity of this pressor region. Because chronic exercise is able to increase cardiovascular health in the hypertensive rat, we hypothesized that exercise-induced caudal hypothalamic plasticity partially underlies the beneficial effects of physical activity. In this review we discuss initial findings from this lab that support this hypothesis. Our experiments demonstrate that chronic exercise alters gene expression and neuronal activity in the caudal hypothalamus of the spontaneously hypertensive rat. These findings describe a potential mechanism by which chronic exercise lowers blood pressure in the hypertensive individual.

Animals↗

PubMed2000

Polyglutamine aggregates alter protein folding homeostasis in Caenorhabditis elegans.

Expansion of polyglutamine repeats in several unrelated proteins causes neurodegenerative diseases with distinct but related pathologies. To provide a model system for investigating common pathogenic features, we have examined the behavior of polyglutamine expansions expressed in Caenorhabditis elegans. The expression of polyglutamine repeats as green fluorescent protein (GFP)-fusion proteins in body wall muscle cells causes discrete cytoplasmic aggregates that appear early in embryogenesis and correlates with a delay in larval to adult development. The heat shock response is activated idiosyncratically in individual cells in a polyglutamine length-dependent fashion. The toxic effect of polyglutamine expression and the formation of aggregates can be reversed by coexpression of the yeast chaperone Hsp104. The altered homeostasis associated with polyglutamine aggregates causes both the sequestration of an otherwise soluble protein with shorter arrays of glutamine repeats and the relocalization of a nuclear glutamine-rich protein. These observations of induced aggregation and relocalization have implications for disorders involving protein aggregation.

Animals↗

PubMed2000

Development of hypoxia-induced Fos expression in rat caudal hypothalamic neurons.

The caudal hypothalamus is an important CNS site controlling cardiorespiratory integration during systemic hypoxia. Previous findings from this laboratory have identified caudal hypothalamic neurons of anesthetized rats that are stimulated during hypoxia. In addition, patch-clamp recordings in an in vitro brain slice preparation have revealed that there is an age-dependent response to hypoxia in caudal hypothalamic neurons. The present study utilized the expression of the transcription factor Fos as an indicator of neuronal depolarization to determine the hypoxic response of caudal hypothalamic neurons throughout postnatal development in conscious rats. Sprague-Dawley rats, aged three to 56 days, were placed in a normobaric chamber circulated with either 10% oxygen or room air for 3h. Following the hypoxic/normoxic exposure period, tissues from the caudal hypothalamus, periaqueductal gray, rostral ventrolateral medulla and nucleus tractus solitarius were processed immunocytochemically for the presence of the Fos protein. There was a significant increase in the density of neurons expressing Fos in the caudal hypothalamus of hypoxic compared to normoxic adult rats that was maintained in the absence of peripheral chemoreceptors. In contrast, no increase in the density of Fos-expressing caudal hypothalamic neurons was observed during hypoxia in rats less than 12 days old. Increases in Fos expression were also observed in an age-dependent manner in the periaqueductal gray, rostral ventrolateral medulla and nucleus tractus solitarius. These results show an increase in Fos expression in caudal hypothalamic neurons during hypoxia in conscious rats throughout development, supporting the earlier in vitro reports suggesting that these neurons are stimulated by hypoxia.

Age Factors↗

PubMed2000

Nidogen is nonessential and not required for normal type IV collagen localization in Caenorhabditis elegans.

Nidogen (entactin) can form a ternary complex with type IV collagen and laminin and is thought to play a critical role in basement membrane assembly. We show that the Caenorhabditis elegans nidogen homologue nid-1 generates three isoforms that differ in numbers of rod domain endothelial growth factor repeats and are differentially expressed during development. NID-1 appears at the start of embryonic morphogenesis associated with muscle cells and subsequently accumulates on pharyngeal, intestinal, and gonad primordia. In larvae and adults NID-1 is detected in most basement membranes but accumulates most strongly around the nerve ring and developing gonad. NID-1 is concentrated under dense bodies, at the edges of muscle quadrants, and on the sublateral nerves that run under muscles. Two deletions in nid-1 were isolated: cg119 is a molecular null, whereas cg118 produces truncated NID-1 missing the G2 collagen IV binding domain. Neither deletion causes overt abnormal phenotypes, except for mildly reduced fecundity. Truncated cg118 NID-1 shows wild-type localization, demonstrating that the G2 domain is not necessary for nidogen assembly. Both nid-1 mutants assemble type IV collagen in a completely wild-type pattern, demonstrating that nidogen is not essential for type IV collagen assembly into basement membranes.

Alternative Splicing↗

PubMed2000

Papilin in development; a pericellular protein with a homology to the ADAMTS metalloproteinases.

Papilin is an extracellular matrix glycoprotein that we have found to be involved in, (1) thin matrix layers during gastrulation, (2) matrix associated with wandering, phagocytic hemocytes, (3) basement membranes and (4) space-filling matrix during Drosophila development. Determination of its cDNA sequence led to the identification of Caenorhabditis and mammalian papilins. A distinctly conserved 'papilin cassette' of domains at the amino-end of papilins is also the carboxyl-end of the ADAMTS subgroup of secreted, matrix-associated metalloproteinases; this cassette contains one thrombospondin type 1 (TSR) domain, a specific cysteine-rich domain and several partial TSR domains. In vitro, papilin non-competitively inhibits procollagen N-proteinase, an ADAMTS metalloproteinase. Inhibiting papilin synthesis in Drosophila or Caenorhabditis causes defective cell arrangements and embryonic death. Ectopic expression of papilin in Drosophila causes lethal abnormalities in muscle, Malpighian tubule and trachea formation. We suggest that papilin influences cell rearrangements and may modulate metalloproteinases during organogenesis.

Amino Acid Sequence↗

PubMed2000

Regulation of the UNC-5 netrin receptor initiates the first reorientation of migrating distal tip cells in Caenorhabditis elegans.

Cell migrations play a critical role in animal development and organogenesis. Here, we describe a mechanism by which the migration behaviour of a particular cell type is regulated temporally and coordinated with over-all development of the organism. The hermaphrodite distal tip cells (DTCs) of Caenorhabditis elegans migrate along the body wall in three sequential phases distinguished by the orientation of their movements, which alternate between the anteroposterior and dorsoventral axes. The ventral-to-dorsal second migration phase requires the UNC-6 netrin guidance cue and its receptors UNC-5 and UNC-40, as well as additional, UNC-6-independent guidance systems. We provide evidence that the transcriptional upregulation of unc-5 in the DTCs is coincident with the initiation of the second migration phase, and that premature UNC-5 expression in these cells induces precocious turning in an UNC-6-dependent manner. The DAF-12 steroid hormone receptor, which regulates developmental stage transitions in C. elegans, is required for initiating the first DTC turn and for coincident unc-5 upregulation. We also present evidence for the existence of a mechanism that opposes or inhibits UNC-5 function during the longitudinal first migration phase and for a mechanism that facilitates UNC-5 function during turning. The facilitating mechanism presumably does not involve transcriptional regulation of unc-5 but may represent an inhibition of the phase 1 mechanism that opposes or inhibits UNC-5. These results, therefore, reveal the existence of two mechanisms that regulate the UNC-5 receptor that are critical for responsiveness to the UNC-6 netrin guidance cue and for linking the directional guidance of migrating distal tip cells to developmental stage advancements.

Animals↗

PubMed2000

Campylobacter contamination of raw meat and poultry at retail sale: identification of multiple types and comparison with isolates from human infection.

Campylobacter species are the major cause of acute bacterial enteritis reported in the United Kingdom, nonetheless many aspects of campylobacteriosis epidemiology remain poorly understood. The aim of this study was to determine the prevalence of Campylobacter jejuni and Campylobacter coli in fresh bovine, ovine, and porcine liver and chicken portions from retail outlets and compare strain subtype distributions with those associated with cases of human campylobacteriosis occurring within the same period and study area. Meat samples were examined by both enrichment culture and direct plating, and Campylobacter isolates were subjected to the same test procedures (identification, serotyping, phagetyping, resistotyping) applied to the clinical strains. Campylobacter species were isolated from 73.2% of 489 samples examined. Chicken exhibited the highest contamination rate (83.3%), followed by lamb (72.9%), pig (71.7%), and ox (54.2%) liver. C. jejuni predominated in chicken (77.3%), lamb (75.0%), and ox (49.0%) liver, and C. coli predominated in pigs' liver (42.4%). Campylobacter fetus was identified in 12.5% of ox liver samples and also in pig and lamb. Of the human isolates, 89.3% were C. jejuni and 10.7% C. coli. The greatest variation in C. jeuni subtypes was observed among the chicken isolates (57 sero/phage-types), followed by human (48 types) and lamb (30 types). A significant proportion of the chicken and lamb isolates shared identical subtypes with the human strains, indicative of their role as potential sources of infection. Almost 30% of samples yielded multiple strains of Campylobacter, a finding that reinforces the epidemiological importance of selecting and testing more than one presumptive isolate per sample.

Animals↗

PubMed1999

Proteolytic processing of Caenorhabditis elegans SQT-1 cuticle collagen is inhibited in right roller mutants whereas cross-linking is inhibited in left roller mutants.

The sqt-1 gene encodes a C. elegans cuticle collagen that when defective can cause dramatic alterations of organismal morphology. Specific antisera were used to examine the assembly of wild-type and mutant SQT-1 in the cuticle. Wild-type SQT-1 chains associate into dimer, tetramer, and higher oligomers that are cross-linked by non-reducible, presumably tyrosine-derived, covalent bonds. The SQT-1 pattern differs from the bulk of cuticle collagens which are found in trimer and larger forms. sqt-1 mutations that cause left-handed helical twisting of animals remove a conserved carboxyl-domain cysteine and inhibit formation of these non-reducible bonds. SQT-1 monomers accumulate and novel trimer-sized products form. A conserved tyrosine immediately adjacent to the affected cysteine suggests that disulfide bond formation is required for this tyrosine to form a cross-link. sqt-1 mutations that cause right-handed helical twisting affect conserved arginines in a predicted cleavage site for a subtilisin-like protease. These mutant SQT-1 molecules retain residues on the amino side of the predicted cleavage site and are larger than wild-type by the amount expected if cleavage failed to occur. The conservation of this site in all nematode cuticle collagens indicates that they are all synthesized as procollagens that are processed by subtilisin-like proteases.

Animals↗

PubMed1999

In vitro responses of neurons in the periaqueductal gray to hypoxia and hypercapnia.

Hypoxia-sensitive neurons in the caudal hypothalamus (CH) have been shown to project to the periaqueductal gray (PAG) which, in turn, sends descending projections to an area of the ventrolateral medulla (VLM) containing neurons inherently excited by hypoxia. The purpose of this study was to determine if neurons in the PAG are excited by hypoxia or hypercapnia in an in vitro environment. Extracellular responses to hypoxia and hypercapnia of neurons located throughout the PAG were recorded in a rat brain slice (400-500 microm thick) preparation. Hypoxic (10% O(2)/5% CO(2)/85% N(2)) and hypercapnic (7% CO(2)/93% O(2)) stimuli were delivered to the tissue through gas bubbled into the brain slice chamber. A majority (39 of 53) of the neurons tested responded to hypoxia. Of these neurons, 92% responded to hypoxia with an increase in firing rate. Neurons in the dorsolateral/lateral regions increased firing rates to a greater extent than neurons located in ventrolateral regions. All neurons tested (n=6) also responded to hypoxia after perfusion of the tissue with a low Ca(2+)/high Mg(2+) solution to block classic synaptic transmission. Only a small proportion (7/33) of neurons tested responded to hypercapnia. These findings indicate that neurons in the periaqueductal gray region of the brain have an inherent responsiveness to hypoxia and, thus, may contribute to the overall coordination of cardiorespiratory responses to systemic hypoxia.

Animals↗