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The ga1 locus of maize confers unilateral cross incompatibility, preventing cross pollination between females carrying the incompatible Ga1-s allele and males not carrying a corresponding compatible allele. To characterize this system at the molecular level, we carried out a transcript profiling experiment in which silks from near isogenic lines carrying the Ga1-s and ga1 alleles were compared. While several differentially expressed genes were identified, only one mapped to the known location of ga1 . This gene is a pectin methylesterase (PME), which we designated as ZmPme3 , and is present and expressed only in Ga1 - s genotypes. While a functional ZmPME3 is not present in the ga1 genotypes examined, a pectin methylesterase gene cluster is found in ga1 genotypes. The gene cluster in W22 contains 58 tandem full-length or partial PME pseudo genes. These data combined with a wealth of previously published data on the involvement of PMEs in pollen tube growth suggest a role for cell wall modification enzymes in the pollen exclusion component of Ga1-s gametophytic incompatibility. Consistent with this role, a third allele which lacks the female function of Ga1-s, Ga1-m , has a mutationally inactivated version of ZmPme3 .

Trichomonas vaginalis is a protozoan parasite that is the causative agent of trichomoniasis, a widespread sexually transmitted disease. In vitro culture of T. vaginalis typically employs a medium supplemented with either maltose or glucose and carbohydrates are considered essential for growth. Although the nature of the carbohydrates utilized by T. vaginalis in vivo is undefined, the vaginal epithelium is rich in glycogen, which appears to provide a source of carbon for the vaginal microbiota. Here, we show that T. vaginalis grows equally well in growth media supplemented with simple sugars or with glycogen. Analysis of conditioned growth medium by thin layer chromatography indicates that growth on glycogen is accompanied by glycogen breakdown to a mixture of products including maltose, glucose, and oligosaccharides. Enzymatic assays with conditioned growth medium show that glycogen breakdown is accomplished via the release of a glucosidase activity having the properties of an α-amylase into the growth medium. Furthermore, we find that released glucosidase activity increases upon removal of carbohydrate from the growth medium, indicating regulation of synthesis and/or secretion in response to environmental cues. Lastly, we show that addition of T. vaginalis glucosidase activity to a growth medium containing glycogen generates sufficient simple sugar to support the growth of lactobacilli which, themselves, are unable to degrade glycogen. Thus, not only does the glucosidase activity likely play an important role in allowing T. vaginalis to secure simple sugars for its own use, it has the potential to impact the growth of other members of the vaginal microbiome., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The objective of this review is to describe current methods and technologies used to decontaminate food animal carcasses in the United States and describe new technologies and methods that are under development. Bacterial reduction during the conversion of muscle to meat has always been an important challenge for the meat processing industry due to the impact on product safety and quality. More intense microbiological testing and improved microbiological methods have led to a greater awareness by industry and government about the levels of pathogenic bacteria on meat carcasses and in meat products. This increased awareness has spurred research and development on new technologies implemented sequentially in the process, aimed at reducing and eliminating bacteria on carcasses and meat products.

Steers (n = 165) of known percentage Brahman (B) and Angus (A) breeding were used to study effects of breed group (A, 3/4A:1/4B, 1/2A:1/2B, 1/4A:3/4B), age-season of feeding (calves fed during the cool season vs yearlings fed during the warm season) and slaughter end point (less than .90, 1.0 to 1.15, 1.27 to 1.40, greater than or equal to 1.5 cm of adjusted fat over the ribeye) on feedlot performance and carcass characteristics. The 1/2B and 3/4B steers had heavier (P less than .05) initial and final feedlot weights than the A and 1/4B steers and higher (P less than .05) unshrunk ADG than the A steers did. Breed types did not differ for feed efficiency. Yearling steers fed in the warm season had higher (P less than .05) unshrunk ADG than calves fed in the cool season, but ADG calculated on an empty-rumen basis did not differ between the two age-seasons of feeding. Calves fed in the cool season were more efficient (P less than .05) than yearlings fed in the warm season when efficiency was expressed on an empty-rumen basis; however, on a live weight basis there was no difference in feed efficiency. No breed group by age-season of feeding interactions on performance were detected. Slaughter end point did not significantly affect feed efficiency on an empty-rumen basis. The 1/2B and 3/4B steers had smaller ribeye areas (REA) per 100 kg hot carcass and lower marbling scores than the 1/4B and A steers. Yearlings fed in the warm season produced heavier carcasses (P less than .05) than calves fed in the cool season. As s.c. fat thickness at slaughter increased, hot carcass weight and numerical yield grade increased, whereas REA per 100 kg of hot carcass decreased. Marbling also increased as fatness increased up to about 1.5 cm subcutaneous fat.

Activities of acidic proteases (cathepsin B + L) and neutral, calcium-dependent proteases (CDP) were quantified to determine whether differences in proteolytic activity could explain differences in meat tenderness among breed types. Steers (n = 32) of known percentage Angus (A) and Brahman (B) breeding were used to establish differences in meat tenderness (A; 3/4A-1/4B; 1/2A-1/2B; 1/4A-3/4B). Samples were removed from the longissimus muscle within 1 h postmortem and within 2 h were frozen for subsequent determination of cathepsin B + L, CDP-I, CDP-II and CDP-inhibitor activities. Warner-Bratzler shear (WBS) was assessed after 1, 5 and 10 d of postmortem aging. Taste panel evaluations, conducted on steaks that were subjected to 5 d of aging, detected no differences. At d 1, WBS did not differ among breed types; however, by d 10 of aging, steaks from Angus steers were more tender (P less than .05) than steaks from 1/2B and 3/4B steers. The Angus and 1/4B steaks had significantly more (P less than .05) cathepsin B + L activity than the 3/4B. The CDP had no relationship with WBS; however, CDP-inhibitor was positively related to d-1 WBS (r = .41, P less than .05). Cathepsin B + L activity was negatively related to WBS at d 10 (r = -.44, P less than .05). These data suggest that differences in meat tenderness among breed types may be explained partially by differences in proteolytic enzyme activity.

Steers (n = 125) of known percentage Angus (A) and Brahman (B) breeding (A = 31, 3/4A:1/4B = 32, 1/2A:1/2B = 31, 1/4A:3/4B = 31) were slaughtered after being fed as calves during the cool period of the year or fed as yearlings during the warm period of the year. Steers were slaughtered at equivalent outside fat thickness as monitored visually and with real-time ultrasound. Warner-Bratzler shear (WBS) force increased and sensory panel tenderness decreased as percentage Brahman increased. Loin muscle characteristics indicated that differences in tenderness between breed groups were not attributed to cold shortening effects or differences in amount or integrity of connective tissue. Fragmentation values suggested that breed group tenderness differences probably resulted from differences in the muscle fiber component. A 10-d postmortem aging study revealed a differential breed group response to postmortem aging, suggesting that breed groups differed in amount and(or) activity of naturally occurring proteolytic enzymes in muscle tissue.

The effect of acute and chronic morphine administration and naloxone-precipitated withdrawal on the binding of [3H]GABA to its receptor sites in rat brain membranes was investigated. Acute morphine (25 mg/kg) administration produced a decrease in the GABA binding in cerebellum, cortex and striatum. This decrease appears to be due to a selective decrease in the number of high-affinity GABA receptor binding sites. In contrast, rats chronically treated with morphine by pellet implantation did not exhibit any changes in GABA receptor binding, except for an increase in pons medulla. However, in rats which were physically dependent, as indicated by naloxone-precipitated withdrawal, GABA binding was decreased significantly in cerebellum and striatum, relative to chronic morphine treatment or placebo pellet controls. This decrease was due to a decrease in the number of low affinity GABA receptor binding sites. Both chronic morphine and naloxone-precipitated withdrawal treatments produced an increase in GABA binding in the pons medulla. These results suggest that morphine may produce some of its effects by modulating GABAergic systems and that high and low affinity GABA receptor sites may play a differential role during various morphine treatments.

The induction of dopaminergic supersensitivity in rats by the administration of haloperidol in their diet for 30 days (CHAL) in three increasing concentrations (7-15 mg/kg/day) was compared to that induced by single daily subcutaneous injections (SCHAL, 0.7 mg/kg) on the basis of biochemical (radioimmunoassay of serum haloperidol levels, 3H-spiroperidol binding) or behavioral (apomorphine stereotypy, spontaneous locomotor activity) parameters. The two modes of administration produced equivalent blood levels of haloperidol by day 30. At 48 hours post treatment: spontaneous locomotor activity and stereotyped behavior were significantly increased in both groups of haloperidol-treated rats, stereotyped behavior was significantly greater in CHAL- vs. SCHAL-treated rats at 8 days post treatment and specific 3H-spiroperidol binding was increased 64% and 236% within the striatum and GP, respectively, of CHAL-treated vs. control rats. Scatchard analysis of 3H-spiroperidol binding isotherms revealed a significant increase in the Bmax of high affinity binding sites [KD approximately 55 pM] within the striatum of both CHAL- and SCHAL-treated rats at 48 hours post treatment. A second, lower affinity site was resolved within the SCHAL-treated group which was not detected within striatal homogenates of CHAL-treated or control rats.

Experiments were conducted to assess the effects of chronic haloperidol (CHAL) administration on gamma-aminobutyric acid (GABA) receptor binding within the rat globus pallidus (GP) and on the responsiveness of individual pallidal neurons to microiontophoretically applied GABA and glycine. Rats were administered haloperidol in their food for 30 days in increasing concentrations and the experiments were conducted 2 days after termination of the haloperidol treatment. GABA receptor binding and neuronal responsiveness to GABA were significantly increased within the GP following CHAL treatment. The mean EC50 value for GABA was significantly decreased in the CHAL-treated rats, but there was no change in the EC50 for glycine. Scatchard analysis of [3H]muscimol binding demonstrated a single high-affinity binding site (Kd = 5 nM) within both control and CHAL-treated rats. The binding capacity (Bmax) of this high-affinity site was significantly increased in CHAL-treated rats without any change in the dissociation constant (Kd) for this site. These results suggest that CHAL administration may lead to a decrease in GABA release by striatopallidal efferents. The results of this study, combined with those of our previous study on SNR neurons, have demonstrated that blockade of striatally mediated dopamine (DA) neurotransmission leads to similar changes in GABAergic mechanisms at the level of the GP and SNR and suggest that DA regulation of the striatopallidal and striatonigral GABAergic pathways need not be differentially organized as has previously been postulated.

Acetylcholine, biogenic amines, and certain amino acids were applied by microiontophoresis to parvicellular and magnocellular red nucleus (RN) neurons of baboon while recording brachium conjunctivum (BC)-evoked and amino acid-evoked unit discharge from these neurons. Glycine, gamma-aminobutyric acid, and beta-alanine were potent depressants of BC-RN synaptic transmission, amino acid-evoked firing, and spontaneous activity of all RN neurons studied. Glycine was clearly more potent than the other 2 depressant amino acids. L-Glutamic and DL-homocysteic acid were strong excitants of all RN neurons tested. Dopamine, noradrenaline, and 5-hydroxytryptamine depressed the excitability of both parvicellular and magnocellular RN neurons; no excitatory effects were observed with these biogenic amines on RN neurons. Acetylcholine increased the rate of firing of spontaneously discharging parvicellular RN neurons and facilitated the amino acid-induced firing of these same neurons. Acetylcholine did not facilitate BC-RN synaptic transmission nor could this transmission be blocked by cholinergic antagonists. Unlike parvicellular RN neurons, the responsiveness of magnocellular neurons was either unaltered by acetylcholine or slightly decreased. These experiments demonstrate a difference in the pharmacologic responsiveness of parvicellular and magnocellular RN neurons to acetylcholine but do not provide evidence for a cholinergic input to RN via the brachium conjunctivum.

The effects of iontophoretically applied substance P (SP) and eledoisin-related peptide (ERP) on the extracellularly recorded activity of spontaneous or D,L-homocysteic acid (DLH)-driven thoracic (T7-T8) spinal neurons, identified by orthodromic activation by the greater splanchnic nerve, were examined in chloralose/urethane anesthetized cats. Of 24 neurons tested, SP and/or ERP (18-120 nA) produced a weak (ca 30%) excitatory effect on 5 neurons, a weak inhibitory effect on 9 neurons, and no effect on the remaining neurons. When compared to the action of either DLH or GABA, we found both the excitatory and inhibitory effects of SP and/or ERP were delayed and prolonged. These data do not support a role for SP as a major neurotransmitter in spinal circuits in which visceral (splanchnic) afferents participate.

The effect of chronic administration of haloperidol for 30 days and its subsequent withdrawal for 0, 4 and 8 days on the binding of [3H]-GABA and [3H]-muscimol to GABA binding sites in rat brain membranes obtained from six discrete regions of the brain and spinal cord was investigated. The chronic administration of haloperidol resulted in a marked increase in the number of GABA binding sites within the substantia nigra, but did not affect GABA binding sites in other regions of rat brain or spinal cord. The increase in GABA binding sites in the substantia nigra was evident for at least 4 days following termination of haloperidol treatment; however, 8 days following withdrawal from haloperidol, when dopaminergic supersensitivity had become maximally expressed, the number of GABA binding sites in the substantia nigra had declined and was the same as that observed in the control rats. These observations suggest that blockade of dopamine receptors in the striatum by haloperidol results in a compensatory decrease in the activity of the GABAergic strionigral system that leads to an increase in the number of GABA binding sites within the substantia nigra. This increase in the number of GABA binding sites is gradually reversed when dopamine is again allowed to interact with its receptors.

An experimental model of morphine dependence, in which rats were made dependent upon morphine by intraventricular infusion, is described. Morphine dependence was assessed and quantified by a series of withdrawal signs that were induced by the intraperitoneal administration of the morphine antagonist naloxone. The infusion of different concentrations of morphine resulted in the production of physical dependence, the severity of which was directly correlated with the concentration of morphine infused. A weak to moderate degree of dependence characterized by such withdrawal signs as teeth chattering, whole-body shakes, and vocalization was produced by infusions of morphine less than 5 micrograms/hr. A strong degree of physical dependence characterized by additional dominant withdrawal signs such as jumping and launching was produced by the infusion of 50 micrograms/hr morphine. The morphine pellet model that most closely approximated this degree of dependence was a three-pellet model in which a single 75-mg morphine pellet was implanted at 48-hr intervals. Abstinence precipitated by removal of the morphine-containing osmotic minipumps was characterized primarily by teeth chattering and whole-body shakes which persisted for at least 48 hr.

This study was conducted in order to compare the effects of microiontophoretically-applied morphine and met-enkephalin (met-ENK) on spontaneous and/or glutamate-evoked activity of single globus pallidus (GP) neurons in locally anesthetized, paralyzed rats. The predominant effect of both morphine and met-ENK was a depression of pallidal neuronal activity. While very few GP neurons were excited by morphine (2/89), a small population of neurons was excited by met-ENK (16/89). Both the inhibitory and excitatory responses produced by morphine and met-ENK could be attenuated by the microiontophoretic application of naloxone. It was also found that morphine and met-ENK did not affect all GP neurons in a similar manner. When applied to the same neurons, morphine elicited depression in 11 of 16 GP neurons which were excited by the application of met-ENK. In contrast, neither of two GP neurons excited by morphine in this study displayed inhibition upon application of met-ENK. Thus the microiontophoresis of morphine and met-ENK to single GP neurons has demonstrated that these two substances can produce opposite effects when applied to the same neurons and suggests that two functionally distinct types of opiate receptor may exist within rat GP, one which mediates the inhibitory effects of morphine and met-ENK, possibly the classical mu (mu) receptor, and one that is preferentially selective to met-ENK and which mediates the excitatory effects of opiates within this region, possibly the classical delta (delta) receptor.

The effects of the microiontophoretic application of dynorphin A-(1-13) (DYN 13) and the benzomorphans ethylketocyclazocine (EKC), bremazocine and MRZ 2549, (kappa) opioid agonists, and of morphine and morphiceptin, (mu) opioid agonists, were compared on spontaneous or glutamate-evoked discharge of globus pallidus (GP) neurons in rat. Our results demonstrate that mu and kappa opioid agonists are able to depress the excitability of pallidal neurons, possibly by interacting with mu and kappa opioid receptor subtypes, respectively. In addition, the mu agonists and dynorphin A-(1-13), but not the benzomorphans, enhanced the excitability of a number of pallidal neurons. We have proposed a presynaptic site as the basis for this opioid-induced excitation, possibly also mediated by a mu opioid receptor. The selectivity of dynorphin A-(1-13) for benzomorphan kappa opioid receptors in the rat GP appears to be low and dynorphin A-(1-13) may elicit effects that are different from those produced by the benzomorphan kappa agonists by virtue of its ability to interact with other opioid receptor subtypes, for example mu opioid receptors.

Electrophysiological properties of the brachium conjunctivum-red nucleus (BC-RN) synaptic system were studied in barbiturate-anesthetized baboon (Papio). Topographic recordings from the mesencephalon demonstrated that most of the BC-evoked activity was restricted to an area histologically identified as red nucleus; however, some brachium-evoked activity was recorded from the surrounding mesencephalic reticular formation. Short latency BC-evoked activity was also recorded from the pons in the region of nucleus reticularis tegmenti pontis. The majority of the BC fibers were found to conduct at a rate of 44 m/sec; a second group of BC fibers with a slower conduction velocity of 23 m/sec was also observed. Brachium-evoked responses recorded from magnocellular and pravicellular RN neurons were short latency responses consistent with monosynaptic activation of these RN neurons by the BC fibers. The BC-RN synaptic system was found to be a very secure synaptic system and could transmit activity at high rates of stimulation with little or no failure. The responsiveness of the BC fibers was found to be similar to that of optic nerve fibers and pyramidal tract fibers, both of which have been characterized as being similar to peripheral A fibers. The responsiveness of the BC-RN synaptic system began to decrease 5 msec after a single or repetitive transmission and was reduced to about 50% of normal responsiveness at 34 msec. This period of reduced postsynaptic responsiveness was associated with a reduction in presynaptic input to RN and suggest that a disfacilitation at the level of the deep cerebellar nuclei may be in part responsible for the subnormal responsiveness of the BC-RN synaptic system.

These studies were performed to evaluate the effects of chronic haloperidol administration on the responsiveness of the pars reticulata neurons of the substantia nigra (SNR) to microiontophoretically applied gamma-aminobutyric acid (GABA) and glycine. Rats were administered haloperidol in their feed (CHAL treatment) for 30 days in increasing concentrations. All experiments were conducted two days after termination of the CHAL treatment. GABA receptor binding and neuronal responsiveness to GABA were significantly increased within the SNR following CHAL treatment. The mean EC50 value for GABA was significantly decreased for SNR neurons in CHAL-treated rats, but there was no change in the EC50 for glycine. Specific [3H]GABA binding (4 nM) was elevated by more than 50% within the SNR. Scatchard analysis of [3H]muscimol binding data revealed that the binding capacities (Bmax) of both high and low affinity GABA binding sites within the SNR were significantly increased without a change in the apparent dissociation constants (Kd) of either site. Although no regional difference in responsiveness to GABA was detected within the SNR of CHAL-treated rats, the spontaneous activity of neurons located within the rostral two-thirds of the nucleus was reduced; however, there was no change in spontaneous activity of neurons located within the caudal one-third. These data provide direct physiological evidence to support the conclusion that the increase in GABA binding within the SNR following CHAL treatment reflects a neuronal supersensitivity to GABA.

The microiontophoretic application of morphine to single globus pallidus neurons in morphine naive rats resulted in a depression of both spontaneous and glutamate-evoked firing of these neurons. Seventy percent of all pallidal neurons on which morphine was tested exhibited some degree of depression of neuronal discharge; no excitatory effect ws observed to morphine on any pallidal neurons. The depressant effects of morphine, but not those of dopamine, could be antagonized by the microiontophoretic application of the morphine antagonist naloxone, indicating that the morphine-induced depression of pallidal neuronal activity was a specific morphine action. This effect of morphine is most likely mediated by the opiate receptors on pallidal neurons that have been proposed to function in enkephalinergic-mediated synaptic transmission.