Effects of immediate TNF-α exposure on phenotype and function of dendritic cells derived from cord blood mono nuclear cells

Objective: Effects of immediate TNF-α exposure on phenotype and function of dendritic cells (DCs) derived from cord blood mono nuclear cells Materials and Methods: Umbilical cord blood MNCs were isolated from healthy mothers and were divided into TNF(+( and TNF (-) groups. Both were cultured using SCF, Flt3L, GM-CSF and IL-4. But, three ng/ml of TNF-α was first added in the culture of TNF(+) group. All cells were cultured for 14 days and matured with TNF-α or LPS for additional four days. Light microscopic and flowcytometric analyses were performed on days 0, 7, and 14 of both cultures. MLR and cytokines assays were used to characterize the function of immature and mature DCs. Results: Co-culture of cord blood monocytes and hematopoietic stem cells led to the production of DCs with a characteristic veiled appearance and were consistent with a DC panel of surface markers. However, immediate exposure to TNF-α enhanced the survival of culturing cells in the first week of culture and produced mature DCs with higher maturation markers and IL-12 production. Addition of TNF-α as a maturation marker led to the production of matured DCs and also certain immature and hematopoietic stem cells with higher level of IL-10 production. Conclusion: This study developed a simple, easy and cost effective way to generate DCs from non fractionating mononuclear cells. It seems that primitive DCs and monocytes in the MNCs are contented in the presence of TNF. This will lead different hematopoitic stem cells to myeloid pathway and results in DCs

A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development

BACKGROUND: Co-culture of embryos with various somatic cells has been suggested as a promising approach to improve embryo development. Despite numerous reports regarding the beneficial effects of epithelial cells from the female genital tract on embryo development in a co-culture system, little is known about the effect of these cells when being cultured under a polarized condition on embryo growth. Our study evaluated the effects of in vitro polarized cells on pre-embryo development. METHODS: Human endometrial tissue was obtained from uterine specimens excised at total hysterectomy performed for benign indications. Epithelial cells were promptly isolated and cultured either on extra-cellular matrix gel (ECM-Gel) coated millipore filter inserts (polarized) or plastic surfaces (non-polarized). The epithelial nature of the cells cultured on plastic was confirmed through immunohistochemistry, and polarization of cells cultured on ECM-Gel was evaluated by transmission electron microscopy (TEM). One or two-cell stage embryos of a superovulated NMRI mouse were then flushed and placed in culture with either polarized or non-polarized cells and medium alone. Development rates were determined for all embryos daily and statistically compared. At the end of the cultivation period, trophectoderm (TE) and inner cell mass (ICM) of expanded blastocysts from each group were examined microscopically. RESULTS: Endometrial epithelial cells cultured on ECM-Gel had a highly polarized columnar shape as opposed to the flattened shape of the cells cultured on a plastic surface. The two-cell embryos cultured on a polarized monolayer had a higher developmental rate than those from the non-polarized cells. There was no statistically significant difference; still, the blastocysts from the polarized monolayer, in comparison with the non-polarized group, had a significantly higher mean cell number. The development of one-cell embryos in the polarized and non-polarized groups showed no statistically significant difference. CONCLUSION: Polarized cells could improve in vitro embryo development from the two-cell stage more in terms of quality (increasing blastocyst cellularity) than in terms of developmental rate

Anthropometric Analysis of the Human Skull for Developmental Left-Sided Asymmetry, New Finding

Background: Facial and cranial asymmetry is common in bilateral vertebrates, particularly human beings. Through years of injecting fillers and performing face-lift operation on patients, we began to observe that more fillers were required on the right side to reach symmetry and also more loose skin and soft tissue was observed on the right side in the majority of patients (Fig. 5). Here, we conducted an analytical study to comprehensively measure human skulls in order to observe and quantify this difference. Methods: We collected forty-one natural skulls available from five medical schools of Tehran province in Iran. Standard photography was conducted from the front, lateral, and superior views. Measurements and statistics were completed using Image J software (National Institutes of Health) for exact comparison of fixed-point distances on the samples. Results: Comparison of right and left dimensions on the skulls revealed several fixed asymmetries. The most significant differences were measurements around the orbit. The distance between the upper orbit point and zygion (zy), zygion to sub-nasal, and orbital areas were significantly larger on the left side. Measuring the anterior gonion�pogonion distance showed a wider mandibular body on the left side. We did not find any considerable differences between any other left-sided and right-sided skull dimensions. Conclusions: Asymmetry of the face and body is a general rule in human anatomy, but correlation and consistency of this asymmetry between the left and right sides is a novel finding based on our measurements. Our findings showed that the orbit was significantly bigger in width, height, and surface area on the left side. We also quantified a narrower mandible on the right side. We hope these findings can be translated to aesthetic surgery practice to make the plastic surgeon and patient more familiar with the patient�s specific anatomy in order to better predict, plan, and successfully implement aesthetic procedures such as injecting fillers or doing peri-orbital procedures. No Level Assigned: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266. © 2019, Springer Science+Business Media, LLC, part of Springer Nature and International Society of Aesthetic Plastic Surgery

Forehead biconvexity enhancement with fat grafting

Background: Frontal biconvexity is a key criterion for an attractive forehead. Fat injection as an effective, safe and reliable method for soft tissue augmentation could be used to enhance forehead contour. We report our experience with combined platelet-rich plasma (PRP) and fat grafting to create or restore frontal biconvexity and to reduce wrinkles. Methods: Fifty-seven females and 4 males (mean age, 40.2 years) underwent the combined PRP and fat injection. Fat mixed with PRP on the ratio of 5:1 was injected in small aliquots in the space between the dermis and underlying frontalis muscle. Patient satisfaction with the cosmetic result was evaluated by a questionnaire using pre- and postoperative photos and a four-point grading scale. Results: All patients who underwent three injection sessions (n = 5) evaluated the esthetic result as excellent. In patients who had two injection sessions (n = 36), 36 evaluated the result as excellent, 50 as good, and the remaining 5 as moderate. In patients who had a single injection session (n = 15), 27 evaluated the result as excellent, 40 as good, and the remaining 5 as moderate. Concerning frontal wrinkles, 25 of 34 (73.5) patients reported good improvement and the remaining 9 (26.5) reported moderate improvement. Conclusion: Fat injection combined with PRP should be considered as an effective tool to improve forehead biconvexity. Level of evidence: Level IV, therapeutic study. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature

The Arrangement of Microtubules in Embryos Derived from Mice Young, Old and Reconstructed Oocytes

Objective: To study the structure and distribution of microtubules in embryos derivedfrom young, old and reconstructed oocytes.Materials and Methods: Embryos obtained from old (50 embryos), young(50 embryos) and reconstructed oocytes (10 embryos) were studied byimmunocytochemistry. The microtubule structures of the embryos were studied byusing fluroscent microscopy with FITC-PI filter and polyclonal antibody against alfatubulin.Results: The spindle structure of MII young oocyte and the obtained embryos werenormal with the suitable condensation. There was no contact between chromosomeand spindle in old Oocytes as well as the obtained embryos, in addition, the spindlewas extended in old group. In reconstructed embryos, thin and scattered filamentswere observed.Conclusion: This study reveals that the arrangement of microtubules inreconstructed embryos was caused by repeating of injection and oocytemanipulation. Also, interactions between karyoblast, cytoplasm and microtubulsmay not be suitable. This may be caused by low fertilization in these oocytes

Detection of Haemophilus influenzae Type B in cerebrospinal fluid of suspected children with meningitis by PCR

Background: A polymerase chain reaction (PCR) assay with primers from "hpd" gene was compared with culture for the detection of Haemophilus influenzae type b (Hib) in CSF samples from children suspected of meningitis. Methods: 300 cerebrospinal fluid (CSF) samples from suspected meningitis cases were studied by culture (Leventhal & Chocolate Agar) and PCR. Results: The latter method could detect Hib in all 5 culture positive and in 2 of 295 culture negative cases, showing sensitivity, specificity, and an accuracy index of 100%, 99% and 99%, respectively. The PCR result was available within a day. Conclusion: This study has shown that PCR is a rapid, sensitive, and specific diagnostic test for Hib from CSF samples. Furthermore, to maximize management of meningitis cases to reduce the morbidity, mortality, and complications of invasive Hib infection a combination of culture and PCR is necessary for the detection of Hib infection. The incidence of Hib meningitis among children in Children's Medical Center in Tehran based on culture and PCR results were 2.4 % and 1.7% respectively. Based on culture results, Hib causes 15.6% of all cases of bacterial meningitis in children