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1.
Int J Mol Med ; 38(3): 679-86, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27460304

ABSTRACT

Chronic wounds represent a significant burden to health services and are associated with patient morbidity. Novel methods to diagnose and/or treat problematic wounds are needed. Interleukin (IL)-15 is a cytokine involved in a number of biological processes and disease states such as inflammation, healing and cancer progression. The current study explores the expression profile of IL-15 and IL-15 receptor α (IL-15Rα) in chronic wounds and its impact on keratinocytes. IL-15 and IL-15Rα expression were examined in healing and non-healing chronic wounds using qPCR and immunohistochemical analysis. The impact of recombinant IL-15 (rhIL-15) on human adult low calcium temperature (HaCaT) keratinocyte growth and migratory potential was further examined. IL-15 transcript expression was slightly, though non-significantly elevated in healing chronic wounds compared with non-healing chronic wounds. IL-15 protein staining was minimal in both subtypes of chronic wounds. By contrast, IL-15Rα transcript and protein expression were both observed to be enhanced in non-healing chronic wounds compared with healing chronic wounds. The treatment of HaCaT cells with rhIL-15 generally enhanced cell growth and promoted migration. Analysis with small molecule inhibitors suggested that the pro-migratory effect of rhIL-15 may be associated with ERK, AKT, PLCγ and FAK signalling. IL-15 may promote healing traits in keratinocytes and the differential expression of IL-15Rα is observed in chronic wounds. Together, this may imply a complex role for this interleukin in wound healing.


Subject(s)
Cell Movement/genetics , Cell Proliferation/genetics , Interleukin-15/genetics , Keratinocytes/metabolism , Cell Line , Cell Movement/drug effects , Cell Proliferation/drug effects , Chronic Disease , Gene Expression Profiling/methods , Humans , Immunohistochemistry , Interleukin-15/metabolism , Interleukin-15/pharmacology , Interleukin-15 Receptor alpha Subunit/genetics , Interleukin-15 Receptor alpha Subunit/metabolism , Keratinocytes/cytology , Recombinant Proteins/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effects , Signal Transduction/genetics , Wound Healing/drug effects , Wound Healing/genetics , Wounds and Injuries/genetics , Wounds and Injuries/metabolism , Wounds and Injuries/pathology
2.
Neuroscience ; 136(2): 457-66, 2005.
Article in English | MEDLINE | ID: mdl-16226387

ABSTRACT

Immunoreactivity for alpha4, beta1 and delta GABAA receptor subunits on neurones in the periaqueductal gray matter was investigated at different stages of the estrous cycle in Wistar rats. Immunostaining for alpha4, beta1 and delta GABAA receptor subunits was present on neurones throughout the periaqueductal gray matter. The numbers of subunit-immunoreactive neurones remained constant during the early phases of the estrous cycle (proestrus to early diestrus) but increased significantly in late diestrus. Dual immunolabeling for the GABA synthesizing enzyme glutamic acid decarboxylase revealed that almost 90% of the subunit-positive cells contained immunoreactivity for glutamic acid decarboxylase. During the early phases of the estrous cycle (proestrus to early diestrus), approximately one third of the glutamic acid decarboxylase-positive population co-localized alpha4, beta1 and delta GABAA receptor subunits. When the number of subunit positive cells increased during late diestrus, the proportion of the glutamic acid decarboxylase-containing population that expressed alpha4, beta1 and delta GABAA receptor subunits almost doubled. We propose that GABAA receptors with the alpha4beta1delta configuration are expressed by GABAergic neurones in the periaqueductal gray matter and that the numbers of cells expressing these subunits are increased in late diestrus in line with falling plasma progesterone levels. Changes in GABAA receptor expression may lead to changes in the excitability of the neural circuitry in the periaqueductal gray matter.


Subject(s)
Estrous Cycle/physiology , Neuronal Plasticity/physiology , Neurons/physiology , Periaqueductal Gray/physiology , Receptors, GABA-A/biosynthesis , gamma-Aminobutyric Acid/physiology , Animals , Female , Glutamate Decarboxylase/metabolism , Immunohistochemistry , Interneurons/physiology , Periaqueductal Gray/cytology , Periaqueductal Gray/metabolism , Progesterone/blood , Rats , Rats, Wistar
3.
Neuroscience ; 131(2): 397-405, 2005.
Article in English | MEDLINE | ID: mdl-15708482

ABSTRACT

In women, the late luteal phase or "premenstrual" period is commonly associated with psychological disturbances, which include mood changes and increased aggression. The underlying cause is unknown but one possibility is that fluctuations in levels of neuroactive steroids precipitate changes in expression of GABA(A) receptor subunits that result in functional changes in inhibitory control systems. The present study investigated the levels of expression of alpha4, beta1 and delta GABA(A) receptor subunits in the periaqueductal gray matter (PAG) in rats and whether plasticity occurs during the oestrous cycle in females. In male rats alpha4, beta1 and delta subunit immunoreactive neurones were present throughout the PAG in similar numbers. In female rats in proestrus, oestrus and early dioestrus, the density of alpha4, beta1 and delta subunit immunoreactive cells was similar to males. However, in late dioestrus, the numbers increased significantly, especially in the dorsolateral PAG, a region which is particularly rich in GABAergic interneurones. These parallel changes may reflect an increase in expression of the alpha4beta1delta GABA(A) receptor subtype. Recombinant alpha4beta1delta receptors, expressed in Xenopus oocytes, exhibited and EC(50) for GABA an order of magnitude lower (2.02+/-0.33 microM; mean+/-S.E.M.) than that found for the most ubiquitous alpha1beta2gamma2 GABA(A) receptor (32.8+/-2.5 microM). Increased expression of alpha4beta1delta GABA(A) receptors in the interneurones of the PAG could render the panic circuitry abnormally excitable by disinhibiting the ongoing GABAergic inhibition. Similar changes in neuronal excitability within the PAG in women consequent to falling steroid levels in the late luteal phase of the menstrual cycle could contribute to the development of pre-menstrual dysphoria.


Subject(s)
Estrus/metabolism , Mesencephalon/metabolism , Protein Subunits/biosynthesis , Receptors, GABA-A/biosynthesis , Animals , Estrus/genetics , Female , Gene Expression Regulation/physiology , Humans , Male , Neural Inhibition/physiology , Protein Subunits/genetics , Protein Subunits/physiology , Rats , Rats, Wistar , Receptors, GABA-A/genetics , Receptors, GABA-A/physiology , Xenopus laevis
4.
Neurosci Lett ; 326(3): 151-4, 2002 Jul 05.
Article in English | MEDLINE | ID: mdl-12095644

ABSTRACT

The dorsal half of the midbrain periaqueductal grey matter (dPAG) functions as a midbrain aversive system. Serotonin exerts anti-aversive effects in the dPAG that are mediated by actions at 5-hydroxytryptamine (5-HT)(1A)- and 5-HT(2A)-receptors. Since at the cellular level, 5-HT(2A)-receptor activation has been shown to evoke excitatory responses in the dPAG, it is possible that anti-aversive 5-HT(2A)-mediated effects are mediated indirectly by activation of inhibitory gamma-aminobutyric acid (GABA)ergic interneurones. In rats, immunoreactivity for 5-HT(2A)-receptors was present on the soma and dendrites of neurones throughout the PAG. Co-localization studies revealed that the majority (>90%) of 5-HT(2A)-receptor-labelled cells also showed immunoreactivity for GABA. These findings may provide an anatomical substrate for 5-HT(2A)-mediated anti-aversive effects in the dPAG.


Subject(s)
Neurons/metabolism , Periaqueductal Gray/cytology , Receptors, Serotonin/metabolism , gamma-Aminobutyric Acid/metabolism , Animals , Immunohistochemistry , Male , Periaqueductal Gray/metabolism , Rats , Rats, Wistar , Receptor, Serotonin, 5-HT2A
6.
J Appl Bacteriol ; 64(5): 409-20, 1988 May.
Article in English | MEDLINE | ID: mdl-3049504

ABSTRACT

A modified lysine decarboxylase broth has been developed which could be used with a Bactometer M123 to differentiate salmonellas from other bacteria by the characteristics of the conductance detection curve. The medium was used in combination with a selenite cystine trimethylamine oxide dulcitol medium to screen 50 strains of salmonellas and 42 strains of other organisms to establish detection curve magnitude and rate values which could be used to identify curves specific to salmonellas. The combination of media detected all salmonellas tested except Salmonella pullorum. The two media were used to screen 100 inoculated product samples with the Bactometer instrument, in parallel with traditional plating procedures, and using various combinations of pre-enrichment and selective enrichment incubation periods. After 24 h pre-enrichment, the Bactometer system detected more positive samples than the conventional plating procedures after pre-enrichment and selective enrichment. It is considered that these media used in parallel in the Bactometer after conventional pre-enrichment could provide a 48 h screening procedure for salmonellas with a sensitivity comparable to present plating procedures.


Subject(s)
Bacteriological Techniques , Food Microbiology , Salmonella/isolation & purification , Carboxy-Lyases/metabolism , Culture Media , Electric Conductivity , Ferrous Compounds , Quaternary Ammonium Compounds , Salmonella/metabolism , Thiosulfates
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