The Effect of Some Antiseptics on Molds and Yeasts Isolated from Wards in Al-Diwaniya Teaching Hospital, Iraq

The study involved 120 samples collected from wards of the Al-Diwaniya Teaching Hospital, Iraq during the period from 5 April to 5 June 2016. The results showed that the percentage of the fungal contamination in the wards was 77/120 (64.1%), highest of the fungal contamination was recorded in the floors and walls and in the air 18(90%), followed by patients (Nose, Ear and Hands) 14(70%) and lowest contamination was in the instruments 3(15%) when compared. A total of 125 fungal isolates were identified and the isolates belong to 9 genera comprising of Aspergillus spp with 52 isolates (mainly A. niger= 24, A. flavus= 19, A. fumigates= 5 and A. ochraceus= 4). Penicillium spp with 17 isolates, Alternaria alternata with 14 isolates, Candida albicans with 11 isolates, Fusarium spp 9 isolates, Mucor spp 7 isolates, Rhizopus stolonifer 7 isolates, Cryptococcus spp 5 isolates and Rhodotorula spp 3 isolates. The highest occurrence and frequency percentages of fungi recorded for A. niger reaching (18.947%) and (19.2%) respectively, followed by A. flavus reaching (16.842%) and (15.2%) respectively with significant differences when compared with other fungi isolated in this study, while the lowest occurrence and frequency percentages recorded for Rhodotorula spp reaching (2.105%) and (2.4%) respectively. The antiseptics (Dettol, Hibitane and Formalin) have high inhibitory activity against the growth of all fungi studied when used at original concentration which was 10% compared with other concentrations 7.5, 5 and 2.5%. Formalin showed highly significant effect in inhibitory activity for the growth of fungi from Dettol and Hibitane, the percentage of inhibition when using Formalin at concentration 10% ranged between (85.2-95.4%) while for Hibitane between (68.1-79.7%) and for Dettol between (40.6-65.8%) at the same concentration

Discordance between GeneXpert assay and conventional drug-susceptibility testing in detecting rifampicin-resistant tuberculosis: A perspective of the line probe assay

Background: Early detection for tuberculosis (TB) and rifampicin-resistant TB (RRTB) is crucial for proper control of this disease. WHO recommended the use of the GeneXpert assay at district level to cover these two public health demands? A study evaluated the diagnostic impact of the GeneXpert assay in detecting TB and RRTB. Odd results were observed in this study in the form of discordance between the GeneXpert assay and the conventional culture and drug-susceptibility testing (DST). Aim of the study To assess the molecular diagnostic validity of the GeneXpert assay when results do not match phenotypic results given by DST. Methods: Pulmonary TB patients with recently detected sputum positive for acid-fast bacilli (AFB) were recruited from random geographical clusters (18 out of 36 primary healthcare districts in the middle five governorates in Iraq) during a 1-year period (November 2013–October 2014). Sputum samples from all enrolled patients were sent for GeneXpert assay testing, culture, and DST. Genotype mycobacterium (GM) from Hain Lifescience (Nehren, Germany) was used to detect non-tuberculosis mycobacteria (NTM) whenever suspected. Those with discordant results regarding the status of RRTB between GeneXpert assay and DST were retested with the line probe assay (LPA). Simple frequency distribution was used to describe study results. Results: Four-hundred ten patients were enrolled, all of whom were culture positive. Only two patients were found negative for TB on GeneXpert assay who were then diagnosed as NTM by LPA (GM). Out of the 408 patients, discordance between GeneXpert and DST regarding the status of rifampicin susceptibility was observed in 17 cases (4%). Nine patients were RR on GeneXpert but rifampicin susceptible (RS) on DST. LPA agreed with GeneXpert assay for all nine cases. Eight patients were RS on GeneXpert but RR on DST. Here, LPA disagreed with GeneXpert assay only in one patient who was found to be RR by LPA. Conclusion: GeneXpert assay is a valid molecular test for TB and RRTB regardless of its discordance with conventional culture and DST

Histological Evaluation of Restylane Lyft Used as a Scaffold for Dental Pulp Regeneration in Non-Infected Immature Teeth in Dogs

Commercially available hyaluronic acid dermal fillers used as a scaffold in regenerative endodontic procedures (REPs) have demonstrated attractive potentials. This study aimed to histologically evaluate the outcome of REPs using Restylane Lyft (HA) as a scaffold. REPs were performed on pulpless, immature roots in dogs (n = 69). The roots were divided into four groups: blood clot (BC), Restylane Lyft (BC + HA), negative control, and positive control. At 13 weeks postoperatively, hard tissue formation, vascularization, the presence of vascularized soft connective tissue and collagen fibers, the degree of inflammation within pulp spaces and/or periapical tissues, and apical closure were evaluated histologically. The vascularization and formation of loosely arranged collagen fibers within the regenerated soft connective tissues were observed significantly more in the BC+HA group (85% and 40%, respectively; p < 0.05) compared to the BC group (54.6% and 9.1%, respectively; p < 0.05). The degree of inflammation was significantly higher in the HA group than in the BC group; moderate to severe inflammatory cell infiltration was seen in 45% and 13.6% of the cases, respectively. The results of the present study suggest that Restylane Lyft combined with a blood clot used as a scaffold may improve the outcomes of REPs in non-infected, pulpless, immature teeth in dogs

Biosensor Design for the Detection of Circulating Tumor Cells Using the Quartz Crystal Resonator Technique

A new mass-sensitive biosensing approach for detecting circulating tumor cells (CTCs) using a quartz crystal resonator (QCR) has been developed. A mathematical model was used to design a ring electrode-based QCR to eliminate the Gaussian spatial distribution of frequency response in the first harmonic mode, a characteristic of QCRs, without compromising the sensitivity of frequency response. An ink-dot method was used to validate the ring electrode fabricated based on our model. Furthermore, the ring electrode QCR was experimentally tested for its ability to capture circulating tumor cells, and the results were compared with a commercially available QCR with a keyhole electrode. An indirect method of surface immobilization technique was employed via modification of the SiO2 surface of the ring electrode using a silane, protein, and anti-EpCAM. The ring electrode successfully demonstrated eliminating the spatial nonuniformity of frequency response for three cancer cell lines, i.e., MCF-7, PANC-1, and PC-3, compared with the keyhole QCR, which showed nonuniform spatial response for the same cancer cell lines. These results are promising for developing QCR-based biosensors for the early detection of cancer cells, with the potential for point-of-care diagnosis for cancer screening

Nontuberculosis Mycobacteria: Isolation from clinical samples in Iraq

Objective/background: Nontuberculosis mycobacteria (NTM), defined as any mycobacterial strain other than Mycobacterium tuberculosis complex, are a diverse group of pathogens that cause a substantive, but often unappreciated worldwide burden of illness. NTM cause illness similar to M. tuberculosis, but generally do not respond to classic tuberculosis (TB) drug regimens. Here, we evaluated GenoType Mycobacterium CM/AS (Hain Lifesciences) for rapid identification of NTM and compared its results with those of other biochemical tests. Methods: During the study interval from February 2015 to August 2015, samples were tested by GenoType Mycobacterium tuberculosis complex (MTBC) for differentiation of MTB complex, and NTM isolates obtained from patients were analyzed with the GenoType Mycobacterium CM assays for common mycobacteria. Results: All samples tested were M. tuberculosis (typical), except samples from sputum that was negative according to Geno Type MTBC results. All isolates were analyzed with the Geno Type Mycobacterium CM (for common mycobacteria) assays, which correctly identified the species as Mycobacterium chelonae, Mycobacterium intracellulare, Mycobacterium kansasii, and Mycobacterium simiae. Conclusion: GenoType testing of Mycobacteria species using GenoType MTBC and GenoType Mycobacterium CM constitutes a reliable, rapid, simple, and easy-to-interpret assay. Moreover, it appears suitable for use in our region, since it identified all mycobacterial species

Extracellular vesicles produced by primary human keratinocytes in response to TLR agonists induce stimulus-specific responses in antigen-presenting cells.

Cells can communicate through the extracellular vesicles (EVs) they secrete. Pathogen associated molecular patterns (PAMPs), alter the biophysical and communicative properties of EVs released from cells, but the functional consequences of these changes are unknown. Characterization of keratinocyte-derived EVs after poly(I:C) treatment (poly(I:C)-EVs) showed slight differences in levels of EV markers TSG101 and Alix, a loss of CD63 and were positive for autophagosome marker LC3b-II and the cytokine IL36γ compared to EVs from unstimulated keratinocytes (control-EVs). Flagellin treatment (flagellin-EVs) led to an EV marker profile like control-EVs but lacked LC3b-II. Flagellin-EVs also lacked IL-36γ despite nearly identical intracellular levels. While poly(I:C) treatment led to the clear emergence of a > 200 nm diameter EV sub-population, these were not found in flagellin-EVs. EV associated IL-36γ colocalized with LC3b-II in density gradient analysis, equilibrating to 1.10 g/mL, indicating a common EV species. Poly(I:C), but not flagellin, induced intracellular vesicles positive for IL-36γ, LC3b-II, Alix and TSG101, consistent with fusion of autophagosomes and multivesicular bodies. Simultaneous rapamycin and flagellin treatment induced similar intracellular vesicles but was insufficient for the release of IL-36γ+/LC3b-II+ EVs. Finally, a qRT-PCR array screen showed eight cytokine/chemokine transcripts were altered (p < 0.05) in monocyte-derived Langerhans cells (LCs) when stimulated with poly(I:C)-EVs while three were altered when LCs were stimulated with flagellin-EVs compared to control-EVs. After independent confirmation, poly(I:C)-EVs upregulated BMP6 (p = 0.035) and flagellin-EVs upregulated CXCL8 (p = 0.005), VEGFA (p = 0.018) and PTGS2 (p = 0.020) compared to control-EVs. We conclude that exogenous signals derived from pathogens can alter keratinocyte-mediated modulation of the local immune responses by inducing changes in the types of EVs secreted and responses in antigen presenting cells

Outcomes after laser ablation in twin-to-twin transfusion syndrome

Aim: Twin-to-Twin Transfusion Syndrome (TTTS) is associated with high perinatal morbidity and mortality in monochorionic twins. Ultrasound Doppler studies of the umbilical arteries (UAD) have a vital role in fetal assessment in multiple pregnancies complicated by TTTS. The Quintero staging is used to grade the severity of the condition.  Methods: The aim of the study was to describe UAD findings and outcomes in a cohort of 78 twin pregnancies treated with laser ablation.  Results: Of the 78 twin pregnancies, 39 women had two surviving babies (50%) and 17 (22%) had a single survivor. The most frequent Quintero stage at diagnosis was Stage three (38%, 30/78), followed by Stage two (32%, 25/78), Stage one (24%, 19/78) and Stage four (5%, 4/78). The Quintero stage was not significantly associated with survival (chi sq 5.31 p=0.151). While 50% of pregnancies had normal UAD at the time of TTTS diagnosis, 50% had at least one abnormal UAD. A normal UAD was not associated with higher survival (68% v 53%, chi sq 3.26 p=0.071).  Conclusion: Laser ablation for TTTS was associated with 50% double survival and 22% single survival. UAD abnormalities or the Quintero stage was not associated with survival after laser ablation.</p

Cellular Uptake of Gold Nanorods in Breast Cancer Cell Lines

Nanosized materials have been proposed for a wide range of biomedical applications, given their unique characteristics. However, how these nanomaterials interact with cells and tissues, as well as how they bio-distribute in organisms, is still under investigation. Differences such as the nanoparticle size, shape, and surface chemistry affect the basic mechanisms of cellular uptake and responses, which, in turn, affects the nanoparticles&rsquo; applicability for biomedical applications. Thus, it is vital to determine how a specific nanoparticle interacts with cells of interest before extensive in vivo applications are performed. Here, we delineate the uptake mechanism and localization of gold nanorods in SKBR-3 and MCF-7 breast cancer cell lines. Our results show both differences and similarities in the nanorod&ndash;cell interactions of the two cell lines. We accurately quantified the cellular uptake of gold nanorods in SKBR-3 and MCF-7 using inductively coupled plasma mass spectrometry (ICP-MS). We found that both cell types use macropinocytosis to internalize bare nanorods that aggregate and associate with the cell membrane. In addition, we were able to qualitatively track and show intracellular nanoparticle localization using transmission electron microscopy. The results of this study will be invaluable for the successful development of novel and &ldquo;smart&rdquo; nanodrugs based on gold nano-structural delivery vehicles, which heavily depend on their complex interactions with single cells