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Biomedical subjects

R G Miller

Publications and source records attributed to R G Miller.

At least 55 records · Page 3Linked to original sources

Blunted pressor and intramuscular metabolic responses to voluntary isometric exercise in multiple sclerosis.

To test the hypothesis that a lower mean arterial pressure (MAP) response during voluntary isometric exercise in multiple sclerosis (MS) is related to a dampened muscle metabolic signal, 9 MS and 11 control subjects performed an isometric dorsiflexor contraction at 30% maximal voluntary contraction until target failure (endurance time). We made continuous and noninvasive measurements of heart rate and MAP (Finapres) and of intramuscular pH and P(i) (phosphorus magnetic resonance spectroscopy) in a subset of 6 MS and 10 control subjects. Endurance times and change in heart rate were similar in MS and control subjects. The decrease in pH and increase in P(i) were less throughout exercise in MS compared with control subjects, as was the change in MAP response. Differences in muscle strength accounted for some of the difference in MAP response between groups. Cardiovascular responses during Valsalva and cold pressor tests were similar in MS and control subjects, suggesting that the blunted MAP response during exercise in MS was not due to a generalized dysautonomia. The dampened metabolic response in MS subjects was not explained by inadequate central muscle activation. These data suggest that the blunted pressor response to exercise in MS subjects may be largely appropriate to a blunted muscle metabolic response and differences in contracting muscle mass.

Adult↗

Improved plating media for the detection of Salmonella species with typical and atypical hydrogen sulfide production.

The Salmonella detection ability of 2 surfactant-supplemental media, xylose-lysine-tergitol (Nia-proof) 4 (XLT4) and Miller-Mallinson (MM) agar, was compared against that of several commonly used plating media. XLT4 and MM appeared to be the most efficient in detecting Salmonella in meat products and food animal environments. MM was superior to XLT4 in detecting those increasingly more prevalent strains of Salmonella possessing weak to ultraweak H2S production characteristics.

Agar↗

Improved detection of nontyphoid and typhoid Salmonellae with balanced agar formulations.

A strategically balanced medium was developed for the improved detection of nontyphoid and typhoid salmonellae. Its balanced sugar (cellobiose, lactose, mannitol, and trehalose) and protein (beef extract and polypeptone peptone) formulation provided Salmonella with a selective growth advantage over non-Salmonella enteric organisms. The formulations promoted the production and detection of H2S production levels that otherwise might be missed with traditional agar formulations. In combination, these advantages contributed to increased sensitivity without the loss of specificity. In comparative studies using 86 samples of meat products (beef, pork, and chicken), the new media, Miller-Mallinson (MM) agar and xylose lysine tergitol (Niaproof) 4 agar, possessed significantly higher sensitivity (P < 0.001) and an improved specificity over bismuth sulfite, hektoen enteric, and xylose lysine desoxycholate agars. However, these samples did not contain nontyphoid salmonellae with weak to ultraweak H2S production characteristics. Modified formulations of MM agar were generally similar to bismuth sulfite and hektoen enteric agars in the identification of four of seven globally diverse strains of Salmonella serotype Typhi. Two of these seven strains were found to produce more readily identifiable black (H2S-positive) colonies on MM agar, whereas one of the seven was not readily detected by any of the media. The improved detection of nontyphoid and typhoid salmonellae attests to the sensitivity of MM agar and to its potentially broad utility in both clinical and food quality laboratories.

Agar↗

Ly-49CB6 NK inhibitory receptor recognizes peptide-receptive H-2Kb.

NK-mediated cytotoxicity involves two families of receptors: activating receptors that trigger lysis of the target cells being recognized and inhibitory receptors specific primarily for MHC I on the target cell surface that can override the activating signal. MHC I molecules on the cell surface can be classified into molecules made stable by the binding of peptide with high affinity or unstable molecules potentially capable of binding high affinity peptide (hence, peptide receptive) and being converted into stable molecules. It has been previously shown that the Ly-49A inhibitory receptor recognizes stable Dd molecules. We show in this study that the inhibitory receptor Ly-49CB6 recognizes peptide-receptive Kb molecules, but does not recognize Kb molecules once they have bound high affinity peptide.

Animals↗

Consensus guidelines for the design and implementation of clinical trials in ALS. World Federation of Neurology committee on Research.

BACKGROUND: In 1994 consensus guidelines were developed for conducting clinical trials in ALS. With growing experience in clinical trials, it has become clear that a number of further guidelines were needed. METHODS: Under the auspices of the World Federation of Neurology Committee on Research, a multinational group of neurologists, statisticians, patient advocates, representatives from the pharmaceutical industry as well as regulatory agencies developed consensus about a number of revisions to the existing guidelines during a 2 day conference in April 1998. RESULTS: Expanded areas of focus include greater protection of patient rights, more detailed guidelines for outcome measures statistical analyses, disclosure of study results and improved interaction between investigators and the corporate sector. COMMENT: Substantial progress has been made in standardizing and improving the quality of clinical trials in ALS through these consensus guidelines.

Amyotrophic Lateral Sclerosis↗

A multicenter, randomized, double-blinded trial of pyridostigmine in postpolio syndrome.

BACKGROUND: Postpoliomyelitis syndrome (PPS) is likely due to degeneration and dysfunction of terminal axons of enlarged postpolio motor units. Age-related decline in growth hormone and insulin-like growth factor (IGF-I) may be a contributing factor. Neuromuscular junction abnormalities and decreased IGF-I levels may respond to the anticholinesterase pyridostigmine, with consequent improvement in strength, fatigue, and quality of life. OBJECTIVES: To determine the effect of pyridostigmine in PPS on health-related quality of life, isometric muscle strength, fatigue, and serum IGF-I levels; and to assess the safety of pyridostigmine in PPS. METHODS: The study was a multicenter, randomized, double-blinded, placebo-controlled trial of a 6-month course of pyridostigmine 60 mg three times per day in 126 PPS patients. The primary data analysis compared mean changes of outcomes between treatment and control groups at 6 months using an intention to treat approach. Secondary analyses included a comparison of outcomes at 6 and 10 weeks, and in compliant patients. RESULTS: The study showed no significant differences in pyridostigmine and placebo-treated patients with regard to changes in quality of life, isometric strength, fatigue, and IGF-I serum levels at 6 months in the primary analysis and in compliant patients. There were no differences in outcomes at 6 and 10 weeks between groups. However, very weak muscles (1 to 25% predicted normal at baseline) were somewhat stronger (p = 0.10, 95% CI of difference -9.5 to 73.3%), and in compliant patients IGF-I was somewhat increased (p = 0.15, 95% CI of difference -6.4 to 44.8 ng/mL) at 6 months with the medication. Pyridostigmine was generally well tolerated. CONCLUSIONS: This study showed no significant differences between pyridostigmine and placebo-treated PPS patients on measures of quality of life, isometric strength, fatigue, and serum IGF-I.

Aged↗

The NKR-P1B gene product is an inhibitory receptor on SJL/J NK cells.

The mouse NKR-P1 family includes at least three genes: NKR-P1A, -B, -C. Neither surface expression nor function of the NKR-P1B gene product has previously been shown. Here, we demonstrate that the SJL/J allele of the NKR-P1B gene product is expressed on SJL/J NK cells, and is recognized by PK136 mAb. Interestingly, the same mAb does not recognize the NKR-P1B gene product of C57BL/6. We have also generated a novel mAb, 1C10, that recognizes an activation receptor on SJL/J NK cells. Activation of the NKR-P1B receptor-inhibited 1C10 mAb induced redirected lysis and recruited SHP-1, indicating that NKR-P1B is an inhibitory receptor. Therefore, the mouse NKR-P1 gene family, like the Ly49 family, includes both activation and inhibitory receptors.

Amino Acid Sequence↗

A role for perforin in activation-induced T cell death in vivo: increased expansion of allogeneic perforin-deficient T cells in SCID mice.

Despite defective granule exocytosis, T cells from mice whose perforin gene was ablated by homologous recombination (pko mice) caused a similar degree of graft-vs-host disease as normal T cells after injection into sublethally irradiated C.B-17 SCID mice. Moreover host spleens contained significantly greater numbers of T cells from pko mice than from wild-type mice following their i.v. injection. This increase could not be explained by persistence of host APCs that were not cleared by defective donor cytotoxic effector cells. The absence of functional perforin-dependent suppressor cells or an altered cytokine profile of donor T cells could also not account for the behavior of pko cells. Spontaneous and Fas-mediated apoptosis of in vivo activated donor T cells were independent of donor origin. However, pko T blasts exhibited less growth inhibition and cell death after reactivation in vitro. The results are compatible with a model of a defective activation-induced cell death (AICD) pathway, controlled by perforin, accounting for the increased expansion of alloreactive pko T cells.

Animals↗

Design, formulation, and evaluation of isocyanatoacrylate copolymer dental adhesives.

Experiments have recently been completed to explore the development of isocyanatoacrylate copolymers as new dental adhesives. A main goal of this work was to test the utility of solubility parameter differences between the candidate adhesives and etched dentin as a predictor of relative bond strength. All candidate adhesive mixtures contained 2-isocyanatoethyl methacrylate (IEM), a selected amount of tri-n-butylborane oxide (TBBO) initiator, and one of 13 methacrylate comonomers. Reactivity ratios were computed for comonomer pairs as indicators of relative reactivity. The concentration of TBBO was optimized for each comonomer mixture to obtain working times of 2-6 min and setting times of 6-10 min. The solubility parameter difference Deltadelta (J/cm(3))(1/2) was calculated for each test mixture with respect to an etched dentin substrate, as an approximation of wetting ability. Using standard techniques for shear bond strength evaluation, mean shear bond strength values ranging between 7-15.5 MPa were obtained for comonomer adhesives in bonding Z-100 composite to treated dentin. Shear bond strength values showed a good correlation (r = -0.612, P </= 0.05) with solubility parameter differences. This study illustrates the usefulness of reactivity ratios and solubility parameters in the design and development of effective dentin bonding agents.

Chemistry, Pharmaceutical↗

Synchronization of estrus and fertility in zebu beef heifers treated with three estrus synchronization protocols.

The effects on estrus and fertility of 3 estrus synchronization protocols were studied in Brahman beef heifers. In Treatment 1 (PGF protocol; n=234), heifers received 7.5 mg, i.m. prostianol on Day 0 and were inseminated after observed estrus until Day 5. Treatment 2 (10-d NOR protocol; n = 220) consisted of norgestomet (NOR; 3 mg, s.c. implant and 3 mg, i.m.) and estradiol valerate (5 mg, i.m.) treatment on Day -10, NOR implant removal and 400 IU, i.m. PMSG on Day 0, and AI after observed estrus through to Day 5. Treatment 3 (14-d NOR+PGF protocol; n = 168) constituted a NOR implant (3 mg, sc) on Day -14, NOR implant removal on Day 0, PGF on Day 16, and AI after observed estrus through to Day 21. All heifers were examined for return to estrus at the next cycle and inseminated after observed estrus. The heifers were then exposed to bulls for at least 21 d. During the period of estrus observation (5 d) after treatment, those heifers treated with the PGF protocol had a lower (P<0.01) rate of estrual response (58%) than heifers treated with the 10-d NOR (87%) or 14-d NOR+PGF (88%) protocol. Heifers treated with the 10-d NOR protocol displayed estrus earlier and had a closer synchrony of estrus than heifers treated with either the PGF or the 14-d NOR+PGF protocol. Heifers treated with the 14-d NOR+PGF protocol had higher (P<0.05) conception and calving rates (51 and 46%) to AI at the induced estrus than heifers treated with the PGF (45 and 27%) or the 10-d NOR (38 and 33%) protocol. Calving rate to 2 rounds of AI was greater (P<0.05) for heifers treated with the 14-d NOR-PGF (50%) protocol than heifers treated with the 10-d NOR (38%) but not the PGF (43%) protocol. Breeding season calving rates were similar among the 3 protocols. The results show that the 14-d NOR+PGF estrus synchronization protocol induced a high incidence of estrus with comparatively high fertility in Brahman heifers.

Animal Husbandry↗

Long-term persistence of IL-2-unresponsive allogeneic T cells in sublethally irradiated SCID mice.

Donor T cells that are activated by host alloantigens initiate graft versus host disease (GVHD) but their long-term fate is poorly understood. The behavior of alloreactive donor T cells was studied in sublethally irradiated SCID mice. Intravenous injection of 10(6) allogeneic lymphocytes caused a severe form of GVHD, characterized by host hematopoietic atrophy. Fifty-fold fewer donor cells did not induce disease and were not simply rejected by radioresistant host mechanisms. Instead, low numbers of allogeneic T cells expanded 20- to 50-fold and remained for >1 year without causing evidence of GVHD. Persistent non-cycling donor cells with an activated phenotype were mainly found in the spleen. Tolerance was inferred by the recovery of host hematopoiesis, despite the presence of donor allogeneic T cells, and the inability of long-term persisting donor T cells to mediate cellular cytotoxicity or proliferate in response to exogenous IL-2 or antigenic stimulation in vitro. The TCR density of long-term persisting donor T cells was down-regulated. These findings suggest that the development of GVHD depends on the magnitude of the initial anti-host response. Subsequently donor cells differentiate, over several months, into a senescent-like state. This behavior questions the rationale for current treatment approaches to GVHD and is of relevance to any clinical situation where chronic T cell activation takes place in the absence of thymic development.

Animals↗

Carrell-Krusen Symposium invited lecture. Clinical trials in motor neuron diseases.

Although there is no truly effective disease-specific therapy for any of the motor neuron diseases, rapid progress in our understanding of the pathophysiology of some of these disorders is being made. In addition to progress in neuroscience, clinical trials of agents that appear to slow the progress of at least one of these diseases, amyotrophic lateral sclerosis, are beginning to show promising results. The first clinical trials in spinal muscular atrophy are currently underway. A number of other developments have raised the quality of clinical trials, which should improve their productivity and efficiency in the future.

Acetates↗

NK cells can recognize different forms of class I MHC.

NK recognition and lysis of targets are mediated by activation receptor(s) whose effects may be over-ridden by inhibitory receptors recognizing class I MHC on the target. Incubation of normal lymphoblasts with a peptide that can bind to their class I MHC renders them sensitive to lysis by syngeneic NK cells. By binding to class I MHC, the peptide alters or masks the target structure recognized by an inhibitory NK receptor(s). This target structure is most likely an "empty" dimer of class I heavy chain and beta2m as opposed to a "full" class I trimer formed by binding of specific peptide that is recognized by CTL.

Amino Acid Sequence↗

A role for perforin in activation-induced cell death.

The granule exocytosis pathway of T cell cytotoxicity is absent in mice whose perforin gene has been ablated by targeted mutagenesis. The ability of activated naive T cells to undergo apoptosis in vitro following reaggregation of the TCR complex with anti-TCR mAbs via a Fas-independent pathway was found to be defective in the absence of perforin. Protection from death was most marked in CD8+ T cells. In wild-type cells, perforin was expressed at the same time that apoptosis occurred, and blockade of perforin expression by either incubation with perforin antisense oligonucleotides or with anti-IL-2 Abs resulted in increased viability of activated T cells. The role of perforin was not via perforin-dependent fratricidal killing. The results suggest a model in which perforin acts internally to cause a form of activation-induced T cell death distinct from that caused by members of the TNFR superfamily.

Animals↗

NK cells from human MHC class I (HLA-B) transgenic mice do not mediate hybrid resistance killing against parental nontransgenic cells.

We have investigated the capacity of human MHC class I HLA-B gene products, HLA-B27, -B7 (fully human), and -B7kb (human-mouse hybrid consisting of the alpha1 and alpha2 domains of HLA-B7, and the alpha3 and cytoplasmic domains of mouse H-2Kb), expressed on mouse NK cells during ontogeny to influence NK recognition of otherwise syngeneic mouse target cells. Despite a high level of surface expression of the transgene (comparable to that of endogeneous H-2DbKb molecules), the direct killing of YAC-1 targets, and the killing of P815 targets in a redirected lysis assay, the NK effectors of these transgenic mice could not mediate hybrid resistance-like killing of nontransgenic C57BL/6 target cells either in vitro or in vivo. Splenocytes from B6-B27 mice could be used to generate CTL lines against a B27-binding peptide, implying that T cells restricted by HLA-B27 developed during ontogeny. NK cells from B6-B27 could lyse B6-B27 Con A lymphoblasts pulsed with Db-binding peptide but not B27-binding peptides. Taken together, our results show that these human HLA-B transgene products cannot function as class I MHC "self" elements for mouse NK cells, even when present throughout ontogeny.

Animals↗

Interleukin-2 reverses the defect in activation-induced apoptosis in T cells from autoimmune lpr mice.

Activated T cells from MRLlpr/lpr (lpr) mice have been shown to be resistant to TCR-induced apoptosis (activation-induced cell death) in vitro. We have found that this resistance is related to a defect in IL-2R alpha (CD25) expression and IL-2 signaling. Following primary activation, splenic T cells from 8-week old lpr mice failed to undergo apoptosis after the TCR was religated upon reculture with plate-bound anti-CD3 mAb. These cells had markedly reduced levels of IL-2 secretion and CD25 expression during primary activation in vitro; however, the cells still progressed through the cell cycle and were capable of cell division following TCR religation. Addition of exogenous IL-2 during the primary activation of 8-week-old lpr T cells overcame the defect in CD25 expression. Strikingly, these cells also became sensitive to apoptosis induction and died when the TCR was religated with anti-CD3 mAb. Viable cell recovery of both the lpr CD4+ and CD8+ subsets, as well as the CD4-CD8- subsets, was dramatically reduced under these conditions. Further investigation also revealed that the defect in activation-induced apoptosis in T cells from lpr mice was age-related. Activated T cells from young lpr mice (5 weeks old) underwent apoptosis in response to TCR ligation; these cells also expressed normal levels of CD25 following primary activation. However, as the mice aged from 5 to 8 weeks, susceptibility to TCR-mediated apoptosis in vitro was progressively lost together with the ability to express CD25. Our results suggest that before the onset of severe lymphoaccumulation, activated T cells from young lpr mice possess the capability to undergo TCR-induced apoptosis despite defective fas expression; IL-2 participates in sensitizing the cells to this death pathway. In older mice, this pathway breaks down and, together with the lack of fas-induced apoptosis, may account for the onset of severe lymphoaccumulation and autoimmunity.

Aging↗