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Staphylococcus haemolyticus (S. haemolyticus) is one of the Coagulase-negative staphylococci (CoNS) that inhabits the skin as a commensal. It is increasingly implicated in opportunistic infections, including diabetic foot ulcer (DFU) infections. In contrast to the abundance of information available for S. aureus and S. epidermidis, little is known about the pathogenicity of S. haemolyticus, despite the increased prevalence of this pathogen in hospitalized patients. We described, for the first time, the pathogenesis of different clinical isolates of S. haemolyticus isolated from DFU on primary human skin fibroblast (PHSF) cells. Virulence-related genes were investigated, adhesion and invasion assays were carried out using Giemsa stain, transmission electron microscopy (TEM), MTT and flowcytometry assays. Our results showed that most S. haemolyticus carried different sets of virulence-related genes. S. haemolyticus adhered to the PHSF cells to variable degrees. TEM showed that the bacteria were engulfed in a zipper-like mechanism into a vacuole inside the cell. Bacterial internalization was confirmed using flowcytometry and achieved high intracellular levels. PHSF cells infected with S.haemolyticus suffered from amarked decrease in viability and increased apoptosis when treated with whole bacterial suspensions or cell-free supernatants but not with heat-treated cells. After co-culture with PBMCs, S. haemolyticus induced high levels of pro-inflammatory cytokines. This study highlights the significant development of S. haemolyticus, which was previously considered a contaminant when detected in cultures of clinical samples. Their high ability to adhere, invade and kill the PHSF cells illustrate the severe damage associated with DFU infections.

Staphylococcus haemolyticus (S. haemolyticus) is one of the Coagulase-negative staphylococci (CoNS) that inhabits the skin as a commensal. It is increasingly implicated in opportunistic infections, including diabetic foot ulcer (DFU) infections. In contrast to the abundance of information available for S. aureus and S. epidermidis, little is known about the pathogenicity of S. haemolyticus, despite the increased prevalence of this pathogen in hospitalized patients. We described, for the first time, the pathogenesis of different clinical isolates of S. haemolyticus isolated from DFU on primary human skin fibroblast (PHSF) cells. Virulence-related genes were investigated, adhesion and invasion assays were carried out using Giemsa stain, transmission electron microscopy (TEM), MTT and flowcytometry assays. Our results showed that most S. haemolyticus carried different sets of virulence-related genes. S. haemolyticus adhered to the PHSF cells to variable degrees. TEM showed that the bacteria were engulfed in a zipper-like mechanism into a vacuole inside the cell. Bacterial internalization was confirmed using flowcytometry and achieved high intracellular levels. PHSF cells infected with S.haemolyticus suffered from amarked decrease in viability and increased apoptosis when treated with whole bacterial suspensions or cell-free supernatants but not with heat-treated cells. After co-culture with PBMCs, S. haemolyticus induced high levels of pro-inflammatory cytokines. This study highlights the significant development of S. haemolyticus, which was previously considered a contaminant when detected in cultures of clinical samples. Their high ability to adhere, invade and kill the PHSF cells illustrate the severe damage associated with DFU infections.

Purpose Our purpose was to compare the efficacy of ‘epithelium-off’ and ‘epithelium-on’ cross-linking (CXL) in treatment of progressive keratoconus. Patients and methods This study included 48 eyes of 26 patients who met our inclusion criteria. The Epi- Off CXL group included 32 eyes of 17 patients, and the Epi-On CXL group included 16 eyes of nine patients. Preoperative assessments of uncorrected and bestcorrected visual acuities, refractive errors, keratometry, and corneal tomography including pachymetry, were compared with the postoperative values. Results Preoperatively, there was a statistically nonsignificant difference between the two groups in all studied variables except for the pachymetry at thinnest location. In the Epi-Off group, there was a significant improvement of uncorrected visual acuity, best-corrected visual acuity, Kmax, and inferior–superior value at the 12-month visit. There was late significant worsening of the back elevation and spherical equivalent at the 12-month visit and also significant thinning of pachymetry at thinnest location associated with significant worsening of the average thickness increase. All other variables showed nonsignificant change (stabilization) at both postoperative visits. In the Epi-On group, there was significant thinning of pachymetry at thinnest location and stabilization of uncorrected corrected visual acuity, best-corrected visual acuity, K1, Kmax, (inferior–superior), Y-coordinate, and front elevation at both postoperative visits, and early stabilization with late worsening of all of other variables. Conclusion The Epi-Off CXL was found to be more superior to Epi-On CXL in terms of stabilization of progressive keratoconus but was inevitably associated with complications related to epithelial debridement.

Purpose Our purpose was to compare the efficacy of ‘epithelium-off’ and ‘epithelium-on’ cross-linking (CXL) in treatment of progressive keratoconus. Patients and methods This study included 48 eyes of 26 patients who met our inclusion criteria. The Epi- Off CXL group included 32 eyes of 17 patients, and the Epi-On CXL group included 16 eyes of nine patients. Preoperative assessments of uncorrected and bestcorrected visual acuities, refractive errors, keratometry, and corneal tomography including pachymetry, were compared with the postoperative values. Results Preoperatively, there was a statistically nonsignificant difference between the two groups in all studied variables except for the pachymetry at thinnest location. In the Epi-Off group, there was a significant improvement of uncorrected visual acuity, best-corrected visual acuity, Kmax, and inferior–superior value at the 12-month visit. There was late significant worsening of the back elevation and spherical equivalent at the 12-month visit and also significant thinning of pachymetry at thinnest location associated with significant worsening of the average thickness increase. All other variables showed nonsignificant change (stabilization) at both postoperative visits. In the Epi-On group, there was significant thinning of pachymetry at thinnest location and stabilization of uncorrected corrected visual acuity, best-corrected visual acuity, K1, Kmax, (inferior–superior), Y-coordinate, and front elevation at both postoperative visits, and early stabilization with late worsening of all of other variables. Conclusion The Epi-Off CXL was found to be more superior to Epi-On CXL in terms of stabilization of progressive keratoconus but was inevitably associated with complications related to epithelial debridement.

Purpose Our purpose was to compare the efficacy of ‘epithelium-off’ and ‘epithelium-on’ cross-linking (CXL) in treatment of progressive keratoconus. Patients and methods This study included 48 eyes of 26 patients who met our inclusion criteria. The Epi- Off CXL group included 32 eyes of 17 patients, and the Epi-On CXL group included 16 eyes of nine patients. Preoperative assessments of uncorrected and bestcorrected visual acuities, refractive errors, keratometry, and corneal tomography including pachymetry, were compared with the postoperative values. Results Preoperatively, there was a statistically nonsignificant difference between the two groups in all studied variables except for the pachymetry at thinnest location. In the Epi-Off group, there was a significant improvement of uncorrected visual acuity, best-corrected visual acuity, Kmax, and inferior–superior value at the 12-month visit. There was late significant worsening of the back elevation and spherical equivalent at the 12-month visit and also significant thinning of pachymetry at thinnest location associated with significant worsening of the average thickness increase. All other variables showed nonsignificant change (stabilization) at both postoperative visits. In the Epi-On group, there was significant thinning of pachymetry at thinnest location and stabilization of uncorrected corrected visual acuity, best-corrected visual acuity, K1, Kmax, (inferior–superior), Y-coordinate, and front elevation at both postoperative visits, and early stabilization with late worsening of all of other variables. Conclusion The Epi-Off CXL was found to be more superior to Epi-On CXL in terms of stabilization of progressive keratoconus but was inevitably associated with complications related to epithelial debridement.

Aims: Few studies have compared the interaction of single and repeated administration of amitriptyline (amit) with the nitrergic system and glutamatergic system in the experimental model of neuropathic pain. We aimed to evaluate theantinociceptive effectofsingleandrepeatedadministrationofamitandtoassesswhetherglutamate preceded inducible nitric oxide synthase (iNOS) inhibition as a mechanism of the analgesic effect of amit in the neuropathic model of pain. Materials and methods: Male Wistar rats were subjected to left sciatic nerve ligation. The effect of single (25mgkg−1) and repeated (10mgkg−1 daily for 3weeks) administration of amit intraperitoneally (i.p.) alone or in combination with aminoguanidine (AG i.p., 100mgkg−1 for 3days, a selective iNOS inhibitor) and MK801 (0.05mgkg−1i.p., NMDA antagonist) on resting paw posture and mechanical hyperalgesia were studied. Glutamate level and iNOS protein expression in hippocampus were detected. Key findings: Single and repeated administration of amit alone or in combination with AG or MK-801 demonstrated a significant decrease in resting pain score and increase in the pain threshold. Both glutamate and nitrite levels decreased in the hippocampi of single and repeated amit+MK-801 groups. Immunohistochemistry showed a marked decrease in iNOS immunoreactivity in rats treated with single and repeated amit+MK-801. Significance: Our results suggest that glutamate-dependent mechanisms are involved in the analgesic responses to amit administration. Importantly, glutamatergic system and its upstream nitrergic system play an important role in the antinociceptive action of amit.

Aims: Few studies have compared the interaction of single and repeated administration of amitriptyline (amit) with the nitrergic system and glutamatergic system in the experimental model of neuropathic pain. We aimed to evaluate theantinociceptive effectofsingleandrepeatedadministrationofamitandtoassesswhetherglutamate preceded inducible nitric oxide synthase (iNOS) inhibition as a mechanism of the analgesic effect of amit in the neuropathic model of pain. Materials and methods: Male Wistar rats were subjected to left sciatic nerve ligation. The effect of single (25mgkg−1) and repeated (10mgkg−1 daily for 3weeks) administration of amit intraperitoneally (i.p.) alone or in combination with aminoguanidine (AG i.p., 100mgkg−1 for 3days, a selective iNOS inhibitor) and MK801 (0.05mgkg−1i.p., NMDA antagonist) on resting paw posture and mechanical hyperalgesia were studied. Glutamate level and iNOS protein expression in hippocampus were detected. Key findings: Single and repeated administration of amit alone or in combination with AG or MK-801 demonstrated a significant decrease in resting pain score and increase in the pain threshold. Both glutamate and nitrite levels decreased in the hippocampi of single and repeated amit+MK-801 groups. Immunohistochemistry showed a marked decrease in iNOS immunoreactivity in rats treated with single and repeated amit+MK-801. Significance: Our results suggest that glutamate-dependent mechanisms are involved in the analgesic responses to amit administration. Importantly, glutamatergic system and its upstream nitrergic system play an important role in the antinociceptive action of amit.

Aims: Few studies have compared the interaction of single and repeated administration of amitriptyline (amit) with the nitrergic system and glutamatergic system in the experimental model of neuropathic pain. We aimed to evaluate theantinociceptive effectofsingleandrepeatedadministrationofamitandtoassesswhetherglutamate preceded inducible nitric oxide synthase (iNOS) inhibition as a mechanism of the analgesic effect of amit in the neuropathic model of pain. Materials and methods: Male Wistar rats were subjected to left sciatic nerve ligation. The effect of single (25mgkg−1) and repeated (10mgkg−1 daily for 3weeks) administration of amit intraperitoneally (i.p.) alone or in combination with aminoguanidine (AG i.p., 100mgkg−1 for 3days, a selective iNOS inhibitor) and MK801 (0.05mgkg−1i.p., NMDA antagonist) on resting paw posture and mechanical hyperalgesia were studied. Glutamate level and iNOS protein expression in hippocampus were detected. Key findings: Single and repeated administration of amit alone or in combination with AG or MK-801 demonstrated a significant decrease in resting pain score and increase in the pain threshold. Both glutamate and nitrite levels decreased in the hippocampi of single and repeated amit+MK-801 groups. Immunohistochemistry showed a marked decrease in iNOS immunoreactivity in rats treated with single and repeated amit+MK-801. Significance: Our results suggest that glutamate-dependent mechanisms are involved in the analgesic responses to amit administration. Importantly, glutamatergic system and its upstream nitrergic system play an important role in the antinociceptive action of amit.

ABSTRACT Introduction: Mirabegron is a novel beta-3 receptor agonist. It has potential antiobesity and antidiabetic effects. The aim of the study was the evaluation of the possible antidiabetic, anti-inflammatory and antioxidant effects of mirabegron as well as its effect on serum leptin and adiponectin in diabetic rats alone and in combination with sitagliptin. Materials and Methods: Five groups of Wistar rats were used. Diabetes was induced in group 2, 3, 4 and 5 using streptozotocin in dose of 100 mg/kg. The third, fourth and fifth group were treated for 20 days with sitagliptin (10 mg/kg), mirabegron (10 mg/kg) and combination of the 2 drugs, respectively. Blood samples were used for determination of blood glucose, serum level of leptin, adiponectin, TNF-α, malondialdehyde (MDA) and glutathione (GSH). Results: Streptozotocin caused induction of type-2 diabetes, increased TNF-α and MDA with reduction in the leptin, adiponectin and GSH. Sitagliptin caused a significant reduction of blood glucose, TNF-α and MDA with significant increase in leptin, adiponectin and GSH. Mirabegron caused a non-significant reduction of blood glucose, TNF-α and MDA with significant increase in the leptin and GSH. There was non-significant difference between the effect of the combination of sitagliptin and mirabegron and the effect of sitagliptin alone. Conclusion: Sitagliptin has a moderate anti-inflammatory and antioxidant effects. Mirabegron has a modest antidiabetic, anti-inflammatory and antioxidant effects and failed to normalize the low serum levels of leptin and adiponectin of streptozotocin induced diabetic rats. The addition of mirabegron to sitagliptin shows no additional or synergistic effects.