Antibacterial Activity of Ethyl Acetate and Cream Formulation of Coleus atropurpureus leaves Against Staphylococcus aureus

Infectious diseases caused by Staphylococcus aureus (Sa) bacteria reaching 70% of cases in Asia and can attack and survive in epithelial cells including endothelial cells. Attempts to overcome the infection caused by Sa by giving antibiotics, which work as antibacterial. Myana (Coleus atropurpureus L. Benth) leaves contain several active compounds that might act as antibacterial agents. This study aimed to analyze the antibacterial activity of Coleus Extract in vitro and to evaluate the formulation of the extract cream against the bacteria causing infection, Sa. Coleus leaves ethyl acetate extract contains alkaloids and tannins, the absorbance of Sa at the 3rd hour was getting smaller, from 0.668 to 0.552, this showed that the longer the incubation time of the extract against bacterial inoculation, the more bacteria died. Our cream formulation with criteria: the pH value, homogeneity, spreadability, adhesion, and protection power. Our cream formulation results were then evaluated based on the standard, with criteria: the pH value, homogeneity, spreadability, adhesion, and protection power. It seems that our cream is per the cream standards. It is semi-solid, brownish-white in color, smells typical of Coleus, and pH 6. Moreover, the cream is homogeneous with 5 cm of spreadability, 7 seconds for adhesion, and colorless. Moreover, we found that Coleus extract cream has better antibacterial activity than positive control with a wound closure time of 7-14 days for extract while more than 14 days for positive control.

Imageless robotic-assisted revision arthroplasty from UKA to TKA

Background and objective It is evident from the national joint registries that numbers of revision knee arthroplasty operations are rising. The aim of this article is to introduce a new robotic-assisted approach in UKA to TKA revision arthroplasty and investigate the alignment accuracy, implant component use and surgery time and to compare it to primary robotic-assisted TKA arthroplasty. Methods This retrospective, case-control study included patients undergoing image-less robotic-assisted revision arthroplasty from UKA to TKA (n = 20) and patients undergoing image-less robotic-assisted primary TKA (control group, n = 20) from 11/2018 to 07/2020. The control group was matched based on the BMI and natural alignment. Comparison of groups was based on postoperative alignment, outlier rate, tibial insert size, lateral bone resection depth, incision-to-wound closure time. All surgeries were performed by a single senior surgeon using the same bi-cruciate stabilizing TKA system. Statistical analysis consisted of parametric t‑testing and Fisher’s exact test with a level of significance of p < 0.05. Results The two groups showed no differences in mean BMI, natural alignment (p > 0.05) and mean overall limb alignment. No outlier was found for OLA and slope analysis. The smallest insert size (9 mm) was used in 70% of the cases in the revision group (n = 14) and in 90% of the cases in the primary group (n = 18, p = 0.24), distal femoral and tibial resection depth showed no statistical difference (p > 0.05). The incision to wound closure time was longer in the revision group but showed no significant difference. Conclusion Image-less robotic-assisted revision arthroplasty from UKA to TKA showed a comparable surgery time, and alignment accuracy in comparison to primary robotic-assisted TKA. Comparable bone preservation and subsequent tibial insert size use was observed for both groups.

Performance comparison of the PFA-200 and Anysis-200: Assessment of bleeding risk screening in cardiology patients

BACKGROUND: Assessment of platelet function is important in the management of patients who are subject to operation as well as at potential risk of hemorrhagic complications. OBJECTIVE: This study aimed to evaluate a new platelet assays (Anysis-Epinephrine, Anysis-ADP) and to compare them with PFA-200 in cardiology visiting patients and inpatients. METHODS: Citrated blood samples were collected from 184 patients for ADP test and 163 patients for EPI test, who visited Korea University Guro Hospital with written consent. The PFA-200 assay gives a test result the closure time (CT) until the blood flow rate decreases to 10%of the initial value, whereas Anysis-200 assay does a blood flow migration distance (MD) until blood flow completely stops. According to the results of PFA closure time (CT), the tested samples were classified as either negative control or positive group. The measurements were simultaneously conducted with two devices and compared. RESULTS: The sensitivity and specificity of Anysis-200 C/EPI kit in comparison to PFA-200 C/EPI kit was 87.5%and 85.7%, respectively. Regarding C/ADP kit, the sensitivity and specificity of Anysis-200 was 96.9%and 87.5%, respectively. In addition, the sums of sensitivity and specificity are greater than 150%for both of EPI and ADP. Also, it was found that likelihood ratio and odd ratio for each assay provide useful additional information. Since the Cohen’s kappa coefficients value between the two devices was relatively high, the equivalence between the two devices was confirmed. CONCLUSIONS: Anysis-200, a novel platelet function analyzer has showed excellent agreements with PFA-200 with high agreement rates and precision. Anysis-200 assay would be useful in assessing bleeding risk management as well as abnormal platelet reactivity at point of care.

Effects of the Btk-Inhibitors Remibrutinib (LOU064) and Rilzabrutinib (PRN1008) With Varying Btk Selectivity Over Tec on Platelet Aggregation and in vitro Bleeding Time

Background: Bruton tyrosine kinase inhibitors (BTKi) are used in B-cell malignancies and in development against various autoimmune diseases. Since Btk is also involved in specific pathways of platelet activation, BTKi might be considered to target platelet GPVI/GPIb-mediated atherothrombosis and platelet FcγRIIA-dependent immune disorders. However, BTKi treatment of patients with B-cell malignancies is frequently associated with mild bleeding events caused possibly by off-target inhibition of Tec. Here, we compared the platelet effects of two novel BTKi that exhibit a high (remibrutinib) or low (rilzabrutinib) selectivity for Btk over Tec.Methods and Results: Remibrutinib and rilzabrutinib were pre-incubated with anticoagulated blood. Platelet aggregation and in vitro bleeding time (closure time) were studied by multiple electrode aggregometry (MEA) and platelet-function analyzer-200 (PFA-200), respectively. Both BTKi inhibited atherosclerotic plaque-stimulated GPVI-mediated platelet aggregation, remibrutinib being more potent (IC50 = 0.03 μM) than rilzabrutinib (IC50 = 0.16 μM). Concentrations of remibrutinib (0.1 μM) and rilzabrutinib (0.5 μM), &gt;80% inhibitory for plaque-induced aggregation, also significantly suppressed (&gt;90%) the Btk-dependent pathways of platelet aggregation upon GPVI, von Willebrand factor/GPIb and FcγRIIA activation stimulated by low collagen concentrations, ristocetin and antibody cross-linking, respectively. Both BTKi did not inhibit aggregation stimulated by ADP, TRAP-6 or arachidonic acid. Remibrutinib (0.1 μM) only slightly prolonged closure time and significantly less than rilzabrutinib (0.5 μM).Conclusion: Remibrutinib and rilzabrutinib inhibit Btk-dependent pathways of platelet aggregation upon GPVI, VWF/GPIb, and FcγRIIA activation. Remibrutinib being more potent and showing a better profile of inhibition of Btk-dependent platelet activation vs. hemostatic impairment than rilzabrutinib may be considered for further development as an antiplatelet drug.

Dynamics of Myosin II Filaments during Wound Repair in Dividing Cells

Wound repair of cell membranes is essential for cell survival. Myosin II contributes to wound pore closure by interacting with actin filaments in larger cells; however, its role in smaller cells is unclear. In this study, we observed wound repair in dividing cells for the first time. The cell membrane in the cleavage furrow, where myosin II localized, was wounded by laserporation. Upon wounding, actin transiently accumulated, and myosin II transiently disappeared from the wound site. Ca2+ influx from the external medium triggered both actin and myosin II dynamics. Inhibition of calmodulin reduced both actin and myosin II dynamics. The wound closure time in myosin II-null cells was the same as that in wild-type cells, suggesting that myosin II is not essential for wound repair. We also found that disassembly of myosin II filaments by phosphorylation did not contribute to their disappearance, indicating a novel mechanism for myosin II delocalization from the cortex. Furthermore, we observed that several furrow-localizing proteins such as GAPA, PakA, myosin heavy chain kinase C, PTEN, and dynamin disappeared upon wounding. Herein, we discuss the possible mechanisms of myosin dynamics during wound repair.

P447 Combined fibrin glue with infliximab therapy of postoperative anastomotic fistula in patients with Crohn’s disease: A Pilot study

Background Postoperative anastomotic fistula is one of the most frequent and potentially life-threatening complications following gastrointestinal resections in patients with Crohn’s disease. The purpose of this study was to report the results of infliximab promoting healing of postoperative anastomotic fistula in patients with Crohn’s disease. Methods In this randomized controlled trial, 37 patients with postoperative fistula were assigned to combined(n=18) or control(n=19) groups and followed up for 6 months. The combined group received usual management protocol for fistula plus an intravenous infliximab therapy for 6 months. The control group received usual fistula protocol. The primary outcome was fistula closure time. The secondary outcome was any infectious complication such as recurrence of ECF, abdominal abscess, pneumonia, infectious diarrhea and other signs of infection. Results Thirty-seven patients with postoperative anastomotic fistula were evaluated. There were18 patients randomized to the combined group who received fibrin glue combined with infliximab therapy. There were 19 patients randomized to the control group who received fibrin glue sealant therapy alone. No significant differences in demographic or clinical characteristics were found between the two groups( P&gt;0.05). The combined group had a shorter closure time than the control group(50.5±11.1 vs 76.2±26.2 days, P=0.011). The combined group did not show higher infectious complications than the control group(P=0.167). Conclusion Infliximab strategy with combining fibrin glue is a safe and effective treatment alternative in the management of postoperative anastomotic fistula in Crohn’s disease, offering the advantage of reduced heal time of fistula.

Micropore Closure Rates following Microneedle Application at Various Anatomical Sites in Healthy Human Subjects

<b><i>Introduction:</i></b> The continuous availability of open micropores is crucial for a successful microneedle (MN) drug delivery strategy. However, micropore lifetime depends on intrinsic skin functional and anatomical characteristics, which vary significantly at different anatomical sites. <b><i>Objective:</i></b> This pilot study explored if differences exist in micropore closure timeframes at 3 anatomical sites – upper arm, volar forearm, and abdomen. <b><i>Methods:</i></b> Healthy subjects (<i>n</i> = 35) self-identifying as Asian (<i>n</i> = 9), Bi-/multiracial (<i>n</i> = 2), Black (<i>n</i> = 9), Latino (<i>n</i> = 6), and White (<i>n</i> = 9) completed the study. The upper arm, volar forearm, and abdomen were treated with MNs; skin impedance and transepidermal water loss (TEWL) were measured at baseline and post-MN to confirm micropore formation. Impedance was measured for 3 days to evaluate micropore lifetime. Measurements of L*, which quantifies the skin lightness/darkness, were made using a tristimulus colorimeter. Micropore lifetime was determined by comparing baseline and post-MN impedance measurements, and micropore closure half-life was predicted using mathematical modeling. <b><i>Results:</i></b> Post-MN increase in TEWL and decrease in impedance were significant (<i>p</i> &#x3c; 0.05), confirming successful micropore formation at all anatomical sites. When data were analyzed according to subject self-identified racial/ethnic groups, the mean micropore closure time at the abdomen (63.09 ± 13.13 h) was longer than the upper arm (60.34 ± 14.69 h) and volar forearm (58.29 ± 16.76 h). The predicted micropore closure half-life at anatomical sites was the abdomen (25.86 ± 14.96 h) ≈ upper arm (23.69 ± 13.67 h) &#x3e; volar forearm (20.2 ± 11.99 h). Differences were not statistically significant between groups. Objective categorization by L* showed that the darker skin may be associated with longer micropore closure time at the abdomen site. <b><i>Conclusions:</i></b> Our results suggest that anatomical site of application may not be a source of significant variability in micropore closure time. These findings may help reduce the number of physiological parameters that need to be explicitly considered when developing drug products to support MN-assisted drug delivery strategies.

Establishment of reference intervals for Platelet Function Analyzer -100 closure time in Algerian adults

Introduction The Platelet Function Analyzer-100 (PFA-100) is a point of care instrument that simulates plug formation under high shear flow. The PFA-100 measures the time required to occlude the aperture in a biochemically active cartridge and is expressed in a term of closure time (CT). In Algeria, the reference values used in clinical laboratories are of Western origin. However, ethnic, genetic, dietary environmental, and diet differences between populations may affect reference intervals. We established the reference intervals of PFA-100 closure times in healthy Algerian adults according to the International Federation of Clinical Chemistry method, and we compared them with those of Western and Asian countries. Material and methods We enrolled 303 healthy blood donors in the study. 218 subjects met inclusion criteria. We analyzed the blood sample on the PFA-100 for CT with both the collagen epinephrine and collagen ADP cartridges. Results The reference intervals of PFA-100 collagen epinephrine CT and PFA-100 collagen ADP CT were 91–207 seconds and 71–144 seconds, respectively. Compared to Western and Asian populations, there were significant differences. The upper limits of CTs were higher for Algerians in this study. Our findings show that many healthy Algerians would be incorrectly identified as having a primary hemostasis abnormality according to the reference intervals of the manufacturer and scientific literature. Conclusion This report provides the first reference intervals for PFA-100 CTs in healthy Algerian adults. These results improve the accuracy of diagnosis and patient care in Algeria.