Effects of Imipenem-containing Niosome nanoparticles against high prevalence methicillin-resistant Staphylococcus Epidermidis biofilm formed.

We aim to assess the antibacterial and anti-biofilm properties of Niosome-encapsulated Imipenem. After isolating Staphylococcus epidermidis isolates and determining their microbial sensitivity, their ability to form biofilms was examined using plate microtiter assay. Various formulations of Niosome-encapsulated Imipenem were prepared using the thin-film hydration method, Minimum Biofilm Inhibitory Concentration (MBIC) and Minimum Inhibitory Concentration (MIC) were determined, and biofilm genes expression was examined. Drug formulations' toxicity effect on HDF cells were determined using MTT assay. Out of the 162 separated S. epidermidis, 106 were resistant to methicillin. 87 MRSE isolates were vancomycin-resistant, all of which could form biofilms. The F1 formulation of niosomal Imipenem with a size of 192.3 ± 5.84 and an encapsulation index of 79.36 ± 1.14 was detected, which prevented biofilm growth with a BGI index of 69% and reduced icaD, FnbA, EbpS biofilms' expression with P ≤ 0.001 in addition to reducing MBIC and MIC by 4-6 times. Interestingly, F1 formulation of niosomal Imipenem indicated cell viability over 90% at all tested concentrations. The results of the present study indicate that Niosome-encapsulated Imipenem reduces the resistance of MRSE to antibiotics in addition to increasing its anti-biofilm and antibiotic activity, and could prove useful as a new strategy for drug delivery.

Epidemiology of Dermatophytosis in Tehran, Iran: A Ten-year Retrospective Study.

BACKGROUND: Dermatophytosis still remains a major public health concern worldwide, particularly in developing countries. This study was undertaken to determine the etiological and epidemiological factors of dermatophyte infections in Tehran, Iran. METHODS: A total of 1530 patients clinically suspected of cutaneous fungal infections were examined in two hospitals over a period of 10 years (2010-2020). Samples were analyzed using direct microscopic examination and culture. Data regarding age, gender, and clinical manifestations were also recorded. RESULTS: Out of 1530 cases examined, dermatophytes were detected in 493 (32.2%) patients. Of these patients, 288 (58.4%) were males and 205 (41.6%) were females. The most affected age group was the 25-44 years old (31.6%). Tinea corporis (n=134) was the most prevalent type of ringworm, followed by tinea cruris (n=131), tinea pedis (n=90), tinea manuum (n=65), tinea unguium (n=29), tinea faciei (n=20), tinea capitis (n=18), and tinea barbae (n=2). Both tinea cruris (P<0.001) and tinea pedis (P=0.002) had a significant association with male gender. As for etiological agents, Trichophyton mentagrophytes (29.0%) was the most frequent isolate, followed by Trichophyton tonsurans (25.8%), Trichophyton rubrum (25.3%), Epidermophyton floccosum (6.9%), Trichophyton verrucosum (4.9%), Microsporum audouinii (4.5%), Microsporum canis (2.0%), and Trichophyton violaceum (1.6%). CONCLUSION: Dermatophytes are still the prevailing causes of fungal infection of the skin, hair, and nails in Iran. Further studies with larger samples sizes and inclusion of diverse locations would yield more accurate results.

Comparison of Double Lateral Osteotomy and Asymmetric Dorsal Hump Reduction in Correction of Crooked Nose.

BACKGROUND: Several methods have been introduced to correct crooked nose during rhinoplasty. This study aimed to compare the final shape of nasal pyramid as well as patients' satisfaction of the outcomes in two different rhinoplasty techniques. METHODS: Participants in this study underwent rhinoplasty with two different techniques of double lateral osteotomy in comparison with asymmetric dorsal hump reduction using rasp. Ninety patients were allocated in two groups by a quadruple block randomization. Patients were compared for the correction of nasal deviation 6 and 12 months after surgery. Their self-rated satisfaction with rhinoplasty outcome was also assessed using a researcher-made questionnaire. RESULTS: Crooked nose correction was performed in 45 patients in each surgery group. Primarily, the mean of nasal deviation in two study groups were relatively similar (159.83±22.37 degree in C-shaped group vs. 11.79±4.98 degree in I-shaped group). The changes in degree of deviation after rhinoplasty were statistically significant in both intervention groups. However, based on the shape of nasal curvature, double lateral osteotomy was superior in long term follow up in I-shaped curvatures. Patients' post-operative satisfaction with their nasal appearance was higher in the group of double lateral osteotomy and they were less interested in re-surgery. CONCLUSION: The two rhinoplasty techniques were not statistically different in terms of changes in nasal deviations correction after the surgery. However, long term changes in I-shaped curvatures were more desirable in group of double lateral osteotomy. Use of double lateral osteotomy was associated with better satisfactory aesthetic outcomes among study participants.

Monosomy 21 seen in live born is unlikely to represent true monosomy 21: a case report and review of the literature.

We report a case of a neonate who was shown with routine chromosome analysis on peripheral blood lymphocytes to have full monosomy 21. Further investigation on fibroblast cells using conventional chromosome and FISH analysis revealed two additional mosaic cell lines; one is containing a ring chromosome 21 and the other a double ring chromosome 21. In addition, chromosome microarray analysis (CMA) on fibroblasts showed a mosaic duplication of chromosome region 21q11.2q22.13 with approximately 45% of cells showing three copies of the proximal long arm segment, consistent with the presence of a mosaic ring chromosome 21 with ring instability. The CMA also showed complete monosomy for an 8.8 Mb terminal segment (21q22.13q22.3). Whilst this patient had a provisional clinical diagnosis of trisomy 21, the patient also had phenotypic features consistent with monosomy 21, such as prominent epicanthic folds, broad nasal bridge, anteverted nares, simple ears, and bilateral overlapping fifth fingers, features which can also be present in individuals with Down syndrome. The patient died at 4.5 months of age. This case highlights the need for additional studies using multiple tissue types and molecular testing methodologies in patients provisionally diagnosed with monosomy 21, in particular if detected in the neonatal period.

Lipid profile components and incident cerebrovascular events versus coronary heart disease; the result of 9 years follow-up in Tehran Lipid and Glucose Study.

OBJECTIVE: To assess the effects of lipid component total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and non-HDL-C on risk of stroke events versus coronary heart disease (CHD). METHODS: The study was conducted on 2620 Iranians, aged ≥ 50 years, free from cardiovascular events at baseline (1999-2001). The multivariable hazard ratios (HRs) for stroke/CHD were calculated for 1 mmol/L change in lipid components, using Cox proportional hazard regression. RESULTS: During 9.1 years of follow-up, 73 and 358 cases of stroke and CHD occurred. We found significant interactions between TC and non-HDL-C with gender in risk prediction of stroke. Among women, multivariate adjusted HRs of ischemic stroke were 1.40 (1.08-1.82), 1.66 (0.71-3.86), 2.27 (0.58-8.91), 1.51 (1.06-2.15) and 1.36 (1.024-1.78) for TC, Ln TG, HDL-C, LDL-C and non-HDL-C respectively, while corresponding HRs of ischemic stroke for men were 0.78 (0.55-1.11), 0.71 (0.33-1.51), 1.04 (0.24-4.47), 0.82 (0.56-1.22), 0.78 (0.55-1.11), respectively. We found no interaction between gender and any of the lipids in risk prediction of incident CHD (p > 0.3). All lipid components were independently associated with CHD in whole population. CONCLUSION: The associations of lipid components on ischemic stroke were modified by gender. Only among female population, TC, LDL-C and non-HDL-C were independently associated with increased risk of ischemic stroke. Regarding CHD events, all lipid components were significant predictors.

High-throughput analysis of chromosome abnormality in spontaneous miscarriage using an MLPA subtelomere assay with an ancillary FISH test for polyploidy.

Chromosome analysis of spontaneous miscarriages is clinically important but is hampered by frequent tissue culture failure and relatively low-resolution analysis. We have investigated replacement of conventional karyotype analysis with a quantitative subtelomere assay performed on uncultured tissue samples, which is based on Multiplex Ligation-Dependent Probe Amplification. This assay is suitable for this purpose as approximately 98% of all observed karyotype abnormalities in spontaneous miscarriages involve copy-number change to one or more subtelomere regions. A pilot study has compared karyotyping and subtelomere analysis on 78 samples. Extensive tissue necrosis accounted for failure of both karyotyping and subtelomere testing in four (5.1%) samples. Excluding these, there were no (0/74) subtelomere test failures compared to 9.5% (7/74) karyotype failures. Twenty-two (30%) whole chromosome aneuploidies and five (6.8%) structural abnormalities were detected using the subtelomere assay. With the exception of three cases of triploidy, all karyotype abnormalities were detected by the subtelomere assay. Following on from this study, a further 100 samples were tested using the subtelomere assay in conjunction with a simple ancillary FISH test using uncultured cells to exclude polyploidy in the event of a normal subtelomere assay result. Except for three necrotic samples, tests results were obtained for all cases revealing 18 abnormalities including one case of triploidy. Taking into consideration the high success rate for the combined MLPA and FISH test results, and the very significant additional advantages of cost-effective, high-throughput batching, and automated, objective analysis, this approach greatly facilitates routine investigation of chromosome abnormalities in spontaneous miscarriage.

Unstable Robertsonian translocations der(13;15)(q10;q10): heritable chromosome fission without phenotypic effect in two kindreds.

Robertsonian translocations (RTs) are amongst the most common chromosome abnormalities, but being essentially balanced are not usually associated with phenotypic abnormality. Despite being dicentric, RTs are almost always transmitted stably through cell division without chromosome breakage. We have investigated spontaneous fission of der(13;15)(q10;q10) chromosomes in eight individuals from two unrelated kindreds with a view to assessing clinical significance and to seek an explanation for the peculiar heritable instability displayed by these chromosomes. In Family 1, fission products were observed in five members in three generations. The instability was observed in cells derived from chorionic villus and lymphocytes. In Family 2, the same phenomenon was observed in amniocytes from two separate pregnancies and maternal blood lymphocytes. Detailed FISH analysis of these RTs showed them to be dicentric with an unremarkable pericentromeric structure. Notably, combined immunofluoresence and FISH analysis showed the presence of the centromere-specific proteins CENP-A and CENP-E, consistent with functional dicentricity in >75% of cells analyzed. The fission products are, therefore, presumed to be the result of sporadic, bipolar kinetochore attachment, anaphase bridging with resultant inter-centromeric breakage in a small proportion of mitoses. None of the eight carriers shows phenotypic abnormality and therefore, for prenatal counseling purposes, there appears to be no increased specific risk associated with this phenomenon.

Molecular distinction between true centric fission and pericentric duplication-fission.

Centromere (centric) fission, also known as transverse or lateral centric misdivision, has been defined as the splitting of one functional centromere of a metacentric or submetacentric chromosome to produce two derivative centric chromosomes. It has been observed in a range of organisms and has been ascribed an important role in karyotype evolution; however, the underlying mechanisms remain unknown. We have investigated four cases of apparent centric fission in humans. Two cases show a missing chromosome 22 or 18 that is replaced by two centric ring products, a third case shows two chromosome-10-derived telocentric chromosomes, whereas a fourth case involves the formation of two chromosome-18-derived isochromosomes. In all four cases, results of gross cytogenetic and fluorescence in situ hybridisation analyses were consistent with a simple centric fission event. However, detailed molecular analyses provided evidence in support of centromere duplication as a predisposing mechanism for the observed chromosomal breakage in two of the cases. Results for the third case are consistent with direct centric fission not involving centromere pre-duplication as the likely mechanism. Insufficient material has precluded the further study of the fourth case. The data provide the first molecular evidence for centromere pre-duplication as a possible mechanism to explain the classically assumed simple "centric fission" events in clinical cytogenetics, karyotype evolution and speciation.

Improved testing for CMT1A and HNPP using multiplex ligation-dependent probe amplification (MLPA) with rapid DNA preparations: comparison with the interphase FISH method.

Charcot-Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP) are the two most common peripheral neuropathies, with incidences of about 1 in 2,500. Several techniques can be used to detect the typical 1.5-Mb duplication or deletion associated with these respective conditions, but none combines simplicity with high sensitivity. MLPA is a new technique for measuring sequence dosage. We have assessed its performance for the detection of the specific 1.5-Mb duplication/deletion by prospectively testing 50 patients referred with differential diagnoses of CMT or HNPP. Probes were designed to evaluate the TEKT3, PMP22, and COX10 genes within the CMT1A/HNPP region. We have compared the results with our existing fluorescence in situ hybridization (FISH) assay, which was performed in parallel. There was concordance of results for 49 patients. Of note, one patient showed an intermediate multiplex ligation-dependent probe amplification (MLPA) result with an abnormal FISH result, which is consistent with mosaicism. The assay works equally well with either purified DNA or rapid DNA preparations made by direct cell lysis. The use of the latter significantly reduces the cost of the assay. MLPA is a sensitive, specific, robust, and cost-effective technique suitable for fast, high-throughput testing and offers distinct advantages over other testing methods.

Pericentromeric euchromatin is conserved in minute human supernumerary chromosomes: a study using cross-species colour segmenting (RxFISH).

Investigation of marker chromosomes is one of the most challenging areas of clinical cytogenetics, especially in the prenatal scenario. A range of techniques including microdissection/reverse painting, SKY and M-FISH are available for the investigation of larger markers (>3 Mb). All these techniques rely on hybridization of unique, homologous sequences with simultaneous suppression of repeat sequences. In contrast, RxFISH is based on hybridization of cross-species syntenic sequences; repeat sequences do not hybridize due to species divergence. We have used RxFISH to analyse a group of the smallest, i.e. minute, supernumerary marker chromosomes. Our results suggest that even the smallest marker chromosomes often contain conserved pericentric euchromatin. More detailed characterization of pericentric genetic content is needed to assess the clinical significance of minute supernumerary markers.

Increased gene dosage at Xq26-q27 is associated with X-linked hypopituitarism.

We have identified a novel interstitial duplication at Xq26.1-q27.3 in a previously reported family with X-linked recessive hypopituitarism [1]. Mapping of the duplication was carried out using interphase FISH analysis of over 60 bacterial genomic clones from Xq25-q28. The proximal and distal breakpoints of the duplication are contained within the 432N13 and 91O18 clones, respectively, and are separated by approximately 9 Mb. Comparison with a recently published 13-Mb duplication in another XH family [2] indicated that the duplication break-points in these families were different. Therefore, we conclude that X-linked hypopituitarism is caused by increased dosage of a gene that is critical for pituitary development and that the causative gene is located within the 9-Mb duplicated region that we have defined.