The Attitude of Egyptian Patients with Autoimmune and Rheumatic Diseases towards Telemedicine.

Background and Objectives: The use of telemedicine (TM) has recently undergone rapid growth and proliferation. Professional stakeholders anticipate that TM will aid in the efficient allocation of limited resources in rheumatology care. The aim of the study was to evaluate the acceptance and willingness of Egyptian patients with autoimmune and rheumatic diseases (ARDs) to incorporate TM into rheumatological care and to assess their requirements and concerns regarding TM. Materials and Methods: A cross-sectional questionnaire-based study was conducted among Egyptian patients with ARDs. The questionnaire covered sociodemographic characteristics, clinical and therapeutic data, attitudes, barriers, and motivators towards TM. Results: The study included 189 patients with ARDs, with a mean age of 37 years (SD = 11.71), and 88.4% were females. Participants were divided into two groups based on their acceptance of TM: the non-acceptant group (133, 70.4%) and the acceptant group (56, 29.6%). There was a significant difference in educational level (p = 0.001), chronic kidney and heart disease (p = 0.008 and 0.014, respectively) and hydroxychloroquine administration (p = 0.037) between the two groups. During the coronavirus disease 2019 (COVID-19) pandemic, 96 (50.8%) of participants used virtual rheumatology consultations, mainly using WhatsApp (64.6%). Approximately 87% would require assistance in operating TM technology. The preference for direct conversation with the rheumatologist and the need for physical examination were the main barriers to teleconsultation. Conclusions: TM is opposed by the vast majority of Egyptian patients with ARDs. They are concerned since it does not include a physical examination and prevents them from undergoing additional procedures such as ultrasound and blood testing. The majority of Egyptian patients with ARDs need help using TM technology, which is the most significant barrier to the spread of TM.

Effect of varicella zoster vaccine vs candida antigen injection in treatment of warts.

Treatment of warts is considered as a big challenge for patients as well as doctors. Immunotherapy represents a promising and successful method of warts treatment. A great attention has been paid for various types of immunotherapeutic agents. One of the immunotherapeutic approaches is intralesional immunotherapy that showed a successful result in treatment of warts. Complete resolution of warts was achieved in many of studied patients, while some of them showed partial response and only few patients showed no response. Our study was based on the previous observations and reports of regression of several types of warts after administration of candida antigen and other new immunotherapeutic antigens. Candida antigen group showed complete clearance in 16 patients (69.6%), partial response in seven patients (30.4%). In VZV vaccine group, complete clearance was observed in 15 patients (65.2%), partial response in eight patients (34.8%). These results showed that the therapeutic response in two groups had a close statistical result and more chances must be given to VZV vaccine specially after its promising and successful results. In conclusion, we presented a novel approach for the treatment of recalcitrant wart using intralesional immunotherapy with Candida antigen and VZV vaccine. VZV vaccine seems to be promising, safe and effective remedy for any type warts mainly plantar warts.

Hypofractionated radiation therapy with temozolomide versus standard chemoradiation in patients with glioblastoma multiforme (GBM): A prospective, single institution experience.

BACKGROUND AND AIM: the study aimed to determine whether hypofractionated radiotherapy (HFRT) with simultaneous and adjuvant temozolomide (TMZ) was feasible and could provide adequate disease control in primary GBM patients with poor prognostic factors including large tumor size, poor performance status, unresectable or multifocal lesions, poor imaging and inflammatory indices. PATIENTS AND METHODS: A total of 93 patients with glioblastoma multiforme were collected and distributed randomly as 1:1.7 of cases to controls; cases or arm (I) received HFRT with 45 Gy in 15 fractions over 3 weeks concurrently with TMZ. Controls or arm (II) received standard conventional fractionation radiotherapy of 60 Gy in 30 fractions over 6 weeks concurrently with TMZ. RESULTS: 35 patients were recruited in arm I while 58 patients in arm II with significant difference in site of GBM, pattern of enhancement, type of surgery, and neutrophil to lymphocyte ratio, while no significant differences in tumor size, focality, responses, progression free survival, and overall survival (OS), only the type of surgery was an independent predictor for OS, no significant difference in the type and degree of toxicity between both arms. CONCLUSION: Our results showed that HFRT with concurrent TMZ is a feasible therapeutic approach in patients with GBM, especially those with poor prognostic factors, assuring high treatment compliance and low toxicity rates. Dose escalation and reduction in overall treatment time are clear advantages of HFRT, while at least the same survival rates as conventional fractionated RT are maintained.

Effects of phytate on thyroid gland of rats intoxicated with cadmium.

Cadmium (Cd) is one of the most dangerous occupational and environmental toxins. The objective of the present study is to examine the potential prophylactic effects of phytic acid (PA) on thyroid hormones of male rats intoxicated with Cd. The male albino rats were divided into five groups: group I (control) was fed with the basal diet, group II was intoxicated with Cd in drinking water, groups III, IV, and V were intoxicated with Cd in drinking water and fed with the diet containing 3.5, 7, and 10 g of PA/kg, respectively. The results indicated that the serum calcium, iron (Fe), and total Fe binding capacity levels and serum T3 and T4 in Cd-treated rats of group II were decreased when compared with the control group, while PA-administered groups with Cd showed a significant improvement when compared with the Cd-treated rats only. Serum thyroid stimulating hormone (TSH) level was significantly increased in Cd-treated rats compared with the control group, while the addition of PA in diet decreased the high levels of TSH. These results indicated a prophylactic effect of PA against Cd-induced toxicity in rats.

Comprehensive overview of how to fade into succinate dehydrogenase dysregulation in cancer cells by naringenin-loaded chitosan nanoparticles.

Mitochondrial respiration complexes play a crucial function. As a result, dysfunction or change is intimately associated with many different diseases, among them cancer. The epigenetic, evolutionary, and metabolic effects of mitochondrial complex IΙ are the primary concerns of our review. Provides novel insight into the vital role of naringenin (NAR) as an intriguing flavonoid phytochemical in cancer treatment. NAR is a significant phytochemical that is a member of the flavanone group of polyphenols and is mostly present in citrus fruits, such as grapefruits, as well as other fruits and vegetables, like tomatoes and cherries, as well as foods produced from medicinal herbs. The evidence that is now available indicates that NAR, an herbal remedy, has significant pharmacological qualities and anti-cancer effects. Through a variety of mechanisms, including the induction of apoptosis, cell cycle arrest, restriction of angiogenesis, and modulation of several signaling pathways, NAR prevents the growth of cancer. However, the hydrophobic and crystalline structure of NAR is primarily responsible for its instability, limited oral bioavailability, and water solubility. Furthermore, there is no targeting and a high rate of breakdown in an acidic environment. These shortcomings are barriers to its efficient medical application. Improvement targeting NAR to mitochondrial complex ΙΙ by loading it on chitosan nanoparticles is a promising strategy.

Mapping the Grounds for Mortalities in Acute Myeloid Leukemia Through Registry Analyses: A Retrospective Cohort Study of Children, Adolescents, and Young Adults Patients.

Background: Our objective was to identify non-malignant factors that contribute to mortality in children, adolescents and young adults, aiming to improve patient follow-up and reduce mortality rates to achieve better survival outcomes. Methods: We analyzed 8,239 acute myeloid leukemia (AML) cases diagnosed between 2000 and 2019 in the USA. Using version 8.4.0.1 of the Surveillance, Epidemiology, and End Results (SEER)*Stat software, we calculated the standardized mortality ratios (SMRs) and 95% confidence intervals (CIs) for each cause of death. Results: Out of the 3,165 deaths observed in the study population, the majority (2,245;70.9%) were attributed to AML itself, followed by non-AML cancers (573; 18.1%) and non-cancerous causes (347; 10.9%). Conclusions: Patients with AML are at a higher risk of developing other types of cancer and granulocyte deficiencies, which increases the risk of death from non-cancerous causes such as infections. Moreover, treatment for AML carries the risk of cardiac problems. AML is commoner in males than females.

Trimodality Therapy in which Concurrent Chemoradiation with Concomitant Boost in Muscle Invasive TCC Urinary Bladder Cancer.

BACKGROUND: Bladder cancer is the most common genitourinary tract malignancy among Egyptian males (16%). bladder sparing therapy can be considered an alternative for patients refusing surgery or are not candidates for surgery. The objective of this study was to determine the safety and feasibility of external-beam irradiation with concomitant boost in muscle invasive bladder cancer and to determine the short-term (1-year) risk of recurrence of bladder cancer. METHODS: Between October 2019 and November 2021, we enrolled 42 patients in Prospective, one arm trial. Eligible patients had pathologically confirmed TCC transitional cell carcinoma of the bladder cT2-4aN0M0, who refused surgery or had contraindications to surgery, and treated conservatively with radiotherapy. All patients underwent maximal TURB before beginning of chemoradiation therapy which was delivered in all patients The patients received radiotherapy dose 45 Gy/25 fractions (1.8 Gy) per fraction to the whole bladder+ 3 cm. with   concurrent cisplatin 20 mg/ m2 over 30 minutes  before radiation on days 1,2,15,16,29, and 30. Additionally, concomitant boost limited to the bladder plus1.5 cm margin was deliverd during the last ten days of the treatment with a minimum 6 h gap between fractions, to a total dose 60 Gy, with the overall treatment time equal to 5 weeks. RESULTS: The median overall survival OS for 42 patients with transitional cell carcinoma( TCC) of bladder treated with 3D conformal radiotherapy 3DCRT and concomitant boost was 28 months, the mean OS was 29.9±1.04, and (95% confidence interval=27.9-32). The one-year OS was 100%, 2-year OS was 81%, and 3-year OS was 26.2%.The mean loco-regional relapse free survival (LRRFS) was 31.6±1.8, 95% CI=28.1-35.1, and the median was 26.5±1.4, the one-year loco-regional RFS was 92.9%, and 2-year (LRRFS) was 66.7%.Acute and late genitourinary toxicity was grade 2 in most of patients and also acute and late toxicity of gastrointestinal was equal or less than grade 1. CONCLUSION: In external radiotherapy for muscle invasive bladder cancer a concomitant boost technique of invasive bladder cancer with shortening of the overall treatment time provides a high probability of local control and overall survival with acceptable toxicity.

Impairment of electron transport chain and induction of apoptosis by chrysin nanoparticles targeting succinate-ubiquinone oxidoreductase in pancreatic and lung cancer cells.

BACKGROUND: Flavonoids may help ameliorate the incidence of the major causes of tumor-related mortality, such as pancreatic ductal adenocarcinoma (PDAC) and lung cancer, which are predicted to steadily increase between 2020 to 2030. Here we compared the effect of chrysin and chrysin nanoparticles (CCNPs) with 5-fluorouracil (5-FLU) on the activity and expression of mitochondrial complex II (CII) to induce apoptosis in pancreatic (PANC-1) and lung (A549) cancer cells. METHODS: Chrysin nanoparticles (CCNPs) were synthesized and characterized, and the IC50 was evaluated in normal, PANC-1, and A549 cell lines using the MTT assay. The effect of chrysin and CCNPs on CΙΙ activity, superoxide dismutase activity, and mitochondria swelling were evaluated. Apoptosis was assessed using flow cytometry, and expression of the C and D subunits of SDH, sirtuin-3 (SIRT-3), and hypoxia-inducible factor (HIF-1α) was evaluated using RT-qPCR. RESULTS: The IC50 of CII subunit C and D binding to chrysin was determined and used to evaluate the effectiveness of treatment on the activity of SDH with ubiquinone oxidoreductase. Enzyme activity was significantly decreased (chrysin < CCNPs < 5-FLU and CCNPs < chrysin < 5-FLU, respectively), which was confirmed by the significant decrease of expression of SDH C and D, SIRT-3, and HIF-1α mRNA (CCNPs < chrysin < 5-FLU). There was also a significant increase in the apoptotic effects (CCNPs > chrysin > 5-FLU) in both PANC-1 and A549 cells and a significant increase in mitochondria swelling (CCNPs < chrysin < 5-FLU and CCNPs > chrysin > 5-FLU, respectively) than that in non-cancerous cells. CONCLUSION: Treatment with CCNPs improved the effect of chrysin on succinate-ubiquinone oxidoreductase activity and expression and therefore has the potential as a more efficient formulation than chemotherapy to prevent metastasis and angiogenesis by targeting HIF-1α in PDAC and lung cancer.

The dogma of cetuximab in advanced squamous cell carcinoma of the head and neck after failure of surgery and radiotherapy: is it true among patients in upper Egypt?

Background and aim: We aimed from the current study to explore the treatment results of cetuximab in combination with a weekly carboplatin and paclitaxel regimen in advanced squamous cell carcinoma of head and neck (HNSCC) after failure of radiotherapy and chemotherapy. Methods: This study was a non-randomised, single arm, phase 2 efficacy study conducted in two oncology centres in upper Egypt, we recruited 31 patients with recurrent HNSCC previously treated with concurrent chemoradiation ± surgery to receive weekly cetuximab, carboplatin and paclitaxel for 18 weeks followed by maintenance cetuximab every 2 weeks for 12 months. All patients underwent intention to treat analysis. Results: The current study revealed a significant reduction of the size of recurrent primary lesion (p < 0.001), without comparable significant reduction of regional lymph nodes (LNs) (p = 0.06), the current overall response rate (ORR) was 83.9%, ≥1-year progression-free survival (PFS) was 58.1%, also surgical intervention was succeeded to salvage 32.3% who did not achieve complete response to the current protocol, the median PFS was 12 months which was significantly affected by tumour site (p = 0.012), programmed death ligand-1 (PDL-1) expression (p = 0.01) and overall response rate (ORR) (p < 0.001). Conclusion: Based on favourable treatment outcomes, including high ORR and disease control rate, improved median PFS and tolerable toxicity profile, the current weekly cetuximab, carboplatin and paclitaxel with 1 year maintenance cetuximab in responding patients is considered a feasible and effective regimen.

Detection of ERG11 gene in fluconazole resistant urinary candida isolates.

Candida species resistant to fluconazole and voriconazole were screened for the presence of ERG11gene by polymerase chain reaction (PCR). Also, the association of this gene with the demonstration of Candida virulence factors; biofilm formation, phospholipase and proteinases activities were studied. A total of 61 Candida isolates were collected from urine specimens. Candida species were identified by API 20 C Aux test. Extracellular phospholipase, secretory aspartyl proteinase and biofilm formation were determined. ERG11 gene was detected by PCR. C. albicans was identified in 34.5%, C. glabrata in 29.5% and C. tropicalis and C. krusei in 18% each. Candida species was resistant to fluconazole and voriconazole in 55.7% and 27.9%, respectively. Seventeen (50%) of fluconazole resistant Candida isolates were sensitive to voriconazole. The most frequently Candida species revealed fluconazole resistance were C. glabrata (47.1%), C. krusei (29.4%), and C. tropicalis and C. albicans (11.8% each). Biofilm formation, phospholipase and proteinase activity were determined in 41.2%, 67.6% and 35.3% of fluconazole resistant Candida isolates, respectively. Erg 11 gene was determined in 82.4% of fluconazole resistant Candida isolates and prominent in C. glabrata (93.75%), followed by C. krusei (90%), C. tropicalis (75%) and C. albicans (25%). Erg 11 gene was detected in 64.7% (11/17) of fluconazole resistant-voriconazole sensitive Candida isolates. Regarding, correlation of Erg11 gene positivity and virulence factors among fluconazole resistant Candida isolates, 34.5% exhibited biofilm formation and 62.1% and 31% showed phospholipase and proteinase activities, respectively. There were statistically significant difference concerning the association of proteinase activities and Erg 11 gene expression among fluconazole resistance Candida isolates (P=0.04). The study emphasizes the high prevalence of Erg11 gene among fluconazole resistant Candida species. There was association between the proteinase activity, fluconazole resistance and the presence of Erg11 among Candida species. Voriconazole maintains better activity towards Candida species and represent an alternative therapy.

Therapeutic potential of chrysin nanoparticle-mediation inhibition of succinate dehydrogenase and ubiquinone oxidoreductase in pancreatic and lung adenocarcinoma.

Pancreatic adenocarcinoma (PDAC) and lung cancer are expected to represent the most common cancer types worldwide until 2030. Under typical conditions, mitochondria provide the bulk of the energy needed to sustain cell life. For that inhibition of mitochondrial complex ΙΙ (CΙΙ) and ubiquinone oxidoreductase with natural treatments may represent a promising cancer treatment option. A naturally occurring flavonoid with biological anti-cancer effects is chyrsin. Due to their improved bioavailability, penetrative power, and efficacy, chitosan-chrysin nano-formulations (CCNPs) are being used in medicine with increasing frequency. Chitosan (cs) is also regarded as a highly versatile and adaptable polymer. The cationic properties of Cs, together with its biodegradability, high adsorption capacity, biocompatibility, effect on permeability, ability to form films, and adhesive properties, are advantages. In addition, Cs is thought to be both safe and economical. CCNPs may indeed be therapeutic candidates in the treatment of pancreatic adenocarcinoma (PDAC) and lung cancer by blocking succinate ubiquinone oxidoreductase.

Study of the inhibitory effects of chrysin and its nanoparticles on mitochondrial complex II subunit activities in normal mouse liver and human fibroblasts.

BACKGROUND: Mitochondrial complex ΙΙ has a unique biological role owing to its participation in both the citric acid cycle and the electron transport chain. Our goal was to evaluate the succinate dehydrogenase and ubiquinone oxidoreductase activity of mitochondrial complex II in the presence of chrysin and chrysin-chitosan nanoparticles. Chrysin chitosan nanoparticles were synthesized and characterized using ultraviolet spectroscopy, Fourier transform-infrared spectroscopy, X-ray diffraction, transmission electron microscopy, scanning electron microscopy, drug release, and zeta potential. The binding affinity of chrysin to complex II subunits was assessed by molecular docking. The IC50 values were measured in a suspension of mouse mitochondria, and the inhibitory effect of chrysin and chrysin chitosan nanoparticles on mitochondrial complex ΙΙ was determined. RESULTS: The free energy of binding between chrysin and complex ΙΙ subunits A, B, C, and D was -4.9, -5, -8.2, and -8.4 kcal/mol, respectively. The characteristic peak of chrysin was confirmed at 348 nm. The chrysin chitosan nanoparticles contained characteristic bands of both chrysin and chitosan. The crystalline nature of chrysin chitosan nanoparticles was confirmed by X-ray powder diffraction measurements showing the characteristic Bragg peaks of (11.2°), (32.2°), (19.6°), (27.6°), and (31.96°). Transmission and scanning electron microscopy revealed their spherical shape and an average particle size of 49.7 ± 3.02 nm. Chrysin chitosan nanoparticles showed a burst release within the initial 2 h followed by a steady release at 8 h. Their zeta potential was positive, between +35.5 and +80 mV. The IC50 of chrysin, chitosan nanoparticles, chrysin chitosan nanoparticles, and 5-fluorouracil was 34.66, 184.1, 12.2, and 0.05 µg/mL, respectively, in adult mice liver and 129, 311, 156, and 8.07 µg/mL, respectively, in normal human fibroblasts. When comparing the inhibitory effects on complex ΙΙ activity, application of the IC50 of chrysin, chitosan nanoparticles, chrysin chitosan nanoparticles, and 5-fluorouracil resulted in 40.14%, 90.9%, 86.7%, and 89% decreases in SDH activity and 70.09%, 86.74%, 60.8%, and 80.23% decreases in ubiquinone oxidoreductase activity in normal adult mice, but 80.9%, 89.06%, and 90% significant decreases in SDH activity, and 90%, 85%, and 95% decreases in ubiquinone reductase after treatment with chrysin, chrysin chitosan nanoparticles, and 5-fluorouracil, in normal human fibroblasts, respectively. CONCLUSIONS: Chrysin and CCNPs exhibit potent inhibitory effects on SDH activity ubiquinone oxidoreductase activity.

Could Concurrent Capecitabine with Hypofractionated Radiotherapy in Elderly Patients with Muscle-Invasive Bladder Cancer be an Option?

BACKGROUND: Repopulation of tumor cells during radiotherapy of transitional cell bladder carcinoma is believed to be a significant cause for treatment failure, and it was reported from clinical observations that the local control rate decreased with a prolonged treatment time, so accelerated hypofractionated radiotherapy with concurrent capecitabine may provide good local control in elderly patients unfit for surgery. The study aimed to evaluate the tolerability and efficacy of hypofractionated radiotherapy with capecitabine in elderly patients with urothelial carcinoma. METHODS: Between October 2019 and September 2021, 30 patients with muscle-invasive bladder cancer staged T2-4aN0M0, underwent transurethral resection of bladder tumor followed by capecitabine (825 mg/m2 orally, 2 times a day) and radiation therapy (55 Gy in 2.2 Gy per fraction). RESULTS: Thirty patients with a median age of 73.5 years (range, 65-85) were included in our study. Most patients had T2N0, and T3N0 (28 patients), furthermore 73.3% had an intermediate-grade tumor, Transurethral resection of bladder tumor was incomplete in 43.3. No grade 4 toxicity was documented. Grade 3 urinary toxicities occurred in two patients requiring hospitalization and temporal radiation cessation. Regarding late toxicities, no grade 3 or 4 toxicity was reported. A complete response was obtained in 56.7% of  patients. After a median follow-up of 16 months, the locoregional control rate was 63%. Overall survival, local failure-free survival, and event-free survival were 100%, 93.3%, 80% and 43.3%, 33.3%, 30% at one and two years respectively. CONCLUSION: Hypofractionated chemoradiation with capecitabine, appears to be an effective and well-tolerated curative treatment strategy in the selected elderly population with urothelial carcinoma.

High Rates of Aminoglycoside Methyltransferases Associated with Metallo-Beta-Lactamases in Multidrug-Resistant and Extensively Drug-Resistant Pseudomonas aeruginosa Clinical Isolates from a Tertiary Care Hospital in Egypt.

BACKGROUND: Multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of Pseudomonas aeruginosa are the leading cause of healthcare-associated infections worldwide. OBJECTIVE: The aim was to identify the resistant phenotypes among P. aeruginosa and to characterize different aminoglycosides and carbapenem resistance genes as major mechanisms of resistance in these isolates, in Theodor Bilharz Research Institute (TBRI), a tertiary care hospital in Cairo, Egypt. METHODS: During a period of 11 months, 42 P. aeruginosa clinical isolates were collected from the microbiology laboratory by routine culture. Antimicrobial sensitivity testing to the aminoglycosides gentamicin and amikacin, and other classes of antibiotics, was performed by a disk diffusion method. Isolates were tested for aminoglycoside resistance genes, aac(6')-lb, aac-(3)-lla, rmtB, rmtC, armA, rmtD, and rmtF, and carbapenemase resistance genes bla NDM, bla VIM, and bla IMP, using conventional PCR. RESULTS: Thirty-three (78.5%) of the clinical P. aeruginosa isolates showed MDR and XDR phenotypes at 42.4% and 57.65%, respectively, and these were included in the study. Aminoglycoside resistance was found in 97%, whereas carbapenem resistance was found in 81% of the isolates phenotypically. Only 59.4% (19/26) of the aminoglycoside-resistant isolates harbored resistance genes; none of the amikacin-susceptible isolates harbored any of the tested aminoglycoside resistance genes. Aminoglycoside resistance genes rmtB, armA, aac(6')-lb, and rmtF were found at rates of 17/33 (51.5%), 3/33 (9%), 2/33 (6%), and 2/33 (6%), respectively, whereas rmtD, acc(3)-II, and rmtC were not detected. Only 40.7% (11/27) of the carbapenem-resistant isolates harbored resistance genes. Carbapenem resistance genes, bla NDM andbla VIM, were found at rates of 7/33 (21.2%) and 6/33 (18.1%), respectively, and bla IMP was not detected. CONCLUSION: Rates of MDR and XDR P. aeruginosa and resistance to aminoglycosides and carbapenems in our setting are high. Methyltransferases and metallo-beta-lactamases are the main mechanisms of resistance to aminoglycosides and carbapenems, respectively. The presence of bla NDM and rmtF in the strains confirms their rapid dissemination in the Egyptian environment.

Molecular Characterization of Carbapenem/Colistin-Resistant Acinetobacter baumannii Clinical Isolates from Egypt by Whole-Genome Sequencing.

PURPOSE: The rise of carbapenem-resistant A. baumannii (CRAB) is considered a public health problem limiting the treatment options. Our current work studied the emergence and mechanisms of colistin-resistance among CRAB isolates in Egypt. MATERIALS AND METHODS: Seventeen clinically recovered A. baumannii were identified and screened for their antimicrobial susceptibilities using VITEK-2 system. Colistin susceptibility was evaluated using broth microdilution, and characterization of carbapenem/colistin resistance determinants was performed using whole-genome sequencing (Illumina MiSeq). RESULTS: About 52.9% (9/17) were colistin-resistant. PCR results revealed that all isolates carried bla OXA-51-like genes, bla OXA-23-like was detected in 82.3% (14/17) and bla NDM in 23.5% (4/17). Two isolates harboured bla GES-35 and bla OXA-23. Furthermore, genome analysis of seven isolates revealed six belonged to international clone 2 (IC2) while the remaining isolate was a singleton (ST158), representing a clone circulating in Mediterranean/Middle Eastern countries. CONCLUSION: The emergence and high incidence of colistin-resistance among CRAB clinical isolates in Egypt are alarming because it further limits therapy options and requires prudent antimicrobial stewardship and stringent infection control measures. Whole-genome sequence analyses suggest that the resistance to colistin was associated with multiple mutations in the pmrCAB genes. The high incidence of the high-risk lineage IC2 harbouring bla OXA-23-like as well as bla NDM is also of concern.

First report of the unique expression of RECAF (receptor for alfa feto-protein) in adult B-NHL/CLL patients.

BACKGROUND: Lymphoproliferative disorders (LPDs) are a heterogeneous group of diseases characterized by an uncontrolled production of monoclonal lymphocytes. RECAF is the receptor for alpha-fetoprotein, which is re-expressed on malignant cells, thus serving as a broad-spectrum tumor marker. METHODS: The current study is a retrospective study carried out on 200 archival bone marrow trephine biopsy specimens [60 normal control (NC), 38 pathological control (PC) and 102 lymphoproliferative diseases (LPD) specimens]. RECAF expression was assessed using immunohistochemistry. RESULTS: The percentage of cells that are positive for RECAF was significantly higher in the LPD group than in the NC group (P=0.007), while there was no significant difference between non-Hodgkin lymphoma (NHL) patients and PC regarding the number of RECAF positive cells (P=0.1). RECAF showed a unique expression pattern among the different subtypes of LPD. None of the hairy cell leukemia (HCL) expressed RECAF, while the highest percentage was seen in follicular lymphoma (FL) and diffuse large B cell lymphoma (DLBCL) (P=0.001). Compared to routine histopathology, RECAF was more sensitive in detecting bone marrow (BM) infiltration in FL, mantle cell lymphoma (MCL), and DLBCL (P=0.01). CONCLUSION: RECAF is significantly expressed in the BM of NHL/chronic lymphocytic leukemia (CLL) patients. RECAF shows a unique expression pattern among the different subtypes of LPD. Furthermore, RECAF may help to detect bone marrow infiltration in lymphoma cells. This may help in the diagnosis, follow-up, and targeting of LPD.

Reliability of phenotypic methods for detection of colistin resistance among carbapenem-resistant Acinetobacter baumannii clinical isolates from Egypt.

INTRODUCTION: Acinetobacter baumannii is a challenging pathogen responsible for serious nosocomial infections. Colistin resistance in carbapenem-resistant A. baumannii strains is a critical health problem as it limits the available therapeutic options. The current work aimed to study the reliability of several phenotypic methods for the detection of colistin resistance among carbapenem-resistant A. baumannii isolates in Egypt. METHODS: A total of 22 carbapenem-resistant A. baumannii isolates were recovered. Colistin minimum inhibitory concentrations (MICs) were determined using broth microdilution (BMD) and compared to agar dilution (AD), automated system (VITEK-2) and gradient test (E-test) and were analyzed by statistical methods. RESULTS: Phenotypic testing showed that nine of 22 isolates (40.9%) were colistin-resistant by BMD and seven of them were also resistant by AD, with the categorical agreement (CA) of 72.7% and essential agreement (EA) of 90.9%. Colistin MIC results ranged from 1-8 µg/mL and 1-32 µg/mL by both AD and BMD respectively. Detection of colistin resistance by gradient test and automated system showed high very major error (VME) rates (40.9%) compared to BMD with a lack of CA between them. AD gave moderate agreement with BMD by 90.9% EA, 72.7% CA and only 9.1% VME. CONCLUSIONS: In delineating colistin breakpoints BMD followed by AD method are defined as the only reliable phenotypic methods for colistin resistance evaluation. More rapid and reliable tests, other than BMD and AD, are required for the convenient detection of colistin resistance in the routine clinical microbiology laboratory daily workflow.

High-risk clones and novel sequence type ST4497 of Klebsiella pneumoniae clinical isolates producing different alleles of NDM-type and other carbapenemases from a single tertiary-care centre in Egypt.

Enterobacteria producing NDM carbapenemases represent a severe diagnostic and therapeutic challenge in healthcare settings. Infections caused by NDM-positive strains are usually associated with high mortality rates and very limited treatment options. A total number of 33 carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates were included in this study, comprising 30 recovered from clinical diagnostic samples and 3 cultured from screening rectal swabs taken at patient admission. Bacterial identification was performed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) and antibiotic susceptibility testing was performed by reference broth microdilution and a commercial automated method. Isolates were investigated for carbapenemase production using the ß-CARBA test, the modified carbapenem inactivation method (mCIM) and, for the 30 clinical isolates, by MALDI-TOF/MS, using the MBT STARⓇ-Carba IVD Kit. Carbapenem resistance genes were characterised by PCR and sequencing. Seven different blaNDM gene variants were identified in 94% of the isolates, whilst three variants of blaOXA-48-like were detected in 27% of the isolates. Most CRKP corresponded to high-risk clones (ST147, ST11 and ST15). Novel ST4497 is reported for the first time in this study as well as the first emergence of K. pneumoniae ST231 producing OXA-232 in Egypt. These results indicate an ongoing evolution of the blaNDM genes in our area and confirm the need for a maintained surveillance system in order to monitor the spread of these mobile blaNDM genes.

Carbapenem-resistant Klebsiella pneumoniae isolates from Egypt containing blaNDM-1 on IncR plasmids and its association with rmtF.

OBJECTIVES: The aim of this study was to characterize carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates recovered from clinical specimens at a tertiary care hospital in Egypt over a period of 15 months. METHODS: Eight CRKP isolates were included in this study. The minimum inhibitory concentrations of different antibiotics were determined by broth microdilution and Etest methods. Multilocus sequence typing was performed. Antibiotic resistance genes were assessed by PCR and DNA sequencing. Plasmid analysis was done by S1 nuclease digestion of whole genomic DNA followed by pulsed-field gel electrophoresis (S1-PFGE). RESULT: Eight carbapenem-resistant NDM-1-producing K. pneumoniae isolates of three different sequence types (ST) were identified (ST147, ST11, and ST17), in which blaNDM-1 was carried by either IncR or untypeable plasmids. Seven out of the eight isolates also contained the rmtF methylase gene. CONCLUSION: This study describes the occurrence of IncR plasmids carrying blaNDM-1 and rmtF in Egypt, raising concerns regarding this type of replicon and its role in the transmission of these resistance determinants.