In vitro-Generated MDSCs Reduce the Pregnancy Complications in an Abortion-Prone Murine Model.

Recurrent spontaneous abortion (RSA) is one of the major pregnancy-related complications. The roles of different immune cells have been studied in pregnancy complications. The current study aimed to investigate myeloid-derived suppressor cells (MDSCs) in a murine abortion model and introduce a therapeutic approach by using in vitro-generated MDSCs in this model. CBA/J × DBA/2 (abortion prone) and CBA/J × Balb/C (normal pregnancy) mice were used. The frequency of granulocytic MDSCs, monocytic MDSCs, and Tregs was checked in the bone marrow and uteroplacental tissue of mice on three gestational days (gd9.5, gd13.5, and gd17.5) using the flow cytometry approach. MDSCs were generated in vitro from bone marrow-isolated cells using GM-CSF and IL-6 cytokines. Abortion-prone mice were injected intravenously with in vitro-generated MDSCs at gd0.5, and pregnancy outcomes were recorded in treated mice. The frequency of G-MDSCs and M-MDSCs in the bone marrow of abortion-prone mice was decreased at gd9.5 (p = 0.026 and p = 0.05, respectively). In uteroplacental tissue, the frequency of G-MDSCs was significantly lower at gd9.5 and gd13.5 (p = 0.001, p = 0.029, respectively), while M-MDSCs only showed decreased number at gd9.5 (p = 0.05) in abortion-prone mice. Injection of in vitro-generated MDSCs resulted in the increased fetus and placenta weights (p = 0.049 and p = 0.012, respectively) but showed no effect on the number of live fetuses and abortion rate. The reduced frequency of both G-MDSCs and M-MDSCs in the bone marrow and at the feto-maternal interface is associated with pregnancy complications. In vitro-generated MDSCs could be considered as a potential approach to reduce these complications.

Introducing a New Method for Purification of Human IL-4 by Substitution of a Single Amino Acid in IL-4 Protein Sequence.

BACKGROUND: It is advantageous to develop an effective purification procedure to produce recombinant protein drugs (rPDs) without any tags. To remove N- or C-terminus tags from the rPDs, several cleavage site-based endopeptidases were used. Separating the endopeptidase enzyme from the rPDs is a time-consuming and costly process. OBJECTIVE: To design and develop a new method for the purification of human interleukin (IL)-4 with potential application for other cytokines. METHODS: Met-like amino acids were substituted at position 120 to reduce the possibility of alteration in the structure of IL-4 and its biological activity. Based on the in silico analysis, isoleucine was chosen as an alternative amino acid, and the M120I mutant IL-4 (mIL-4) model was selected for the downstream analysis. Recombinant mIL-4 was produced in the E.coli BL21 host and purified with CNBr. Then in vitro evaluations of the native and mutant IL-4 were performed. RESULTS: The results showed that both the native and mutant IL-4 had the same effect on TF-1 cell proliferation. On the other hand, there was no significant difference between the effects of native IL-4 (nIL-4) and mIL-4 on the expression of IL-4 and IL-10 in activated peripheral blood mononuclear cells. Native and mutant IL-4 have similar biological activities. CONCLUSION: Here, an efficient and straightforward system is introduced to purify IL-4 cytokine using CNBr, which could be applied to other rPDs.

68Ga-DOTATATE Radioisotope scan to detect neuroendocrine tumors; A Cross-Sectional Study.

OBJEVTIVES: Neuroendocrine tumors are a heterogeneous group of neoplasms that arise from the peptide-producing cells of the neuroendocrine system. Different functional imaging methods have been suggested to diagnose NETs. There is still not enough evidence to recommend 68Ga-DOTATATE as a standard diagnostic tool in NETs. Therefore, the aim of this study was to assess the value of 68Ga-DOTATATE scan in detecting NETs. METHODS: This was a cross-sectional study. All patients with a pathologically confirmed NET tumor referred to Masih Daneshvari Hospital affiliated to Shahid Beheshti University of Medical Sciences entered the study. Patients underwent a 68Ga-DOTATATE PET/CT. All statistical analysis were performed by SPSS software version 18. RESULTS: Forty patients with a mean age of 48.1±15.80 years entered the study. Twenty-one (52.5%) were male and 19 (47.5%) female. In the studied patients, neuroendocrine tumor was present in 19 cases (47.5%) in pancreas and gastrointestinal tract, 9 (22.5%) in lung, 3 (7.5%) in mediastinum and adrenal gland, 6 cases (5%) in liver and 3 other sites. There was no significant association between mean age and gender with primary location of the tumor. The mean SUVmax was 11.62±20.02 and the the mean tumor size was 38.25±31.35 mm. The mean size of the metastasis was 40.55±24.53 mm. The mean percentage of ki-67 was 12.54±18.40. There was no significant correlation between SUVmax of the lesion and age (r=0.063, P=0.701), tumor size (r=-0.63, P=0.067) or Ki-67 (r=0.011, P=0.960). In 20 cases, metastases were reported, of which 14 were (70%) in the liver, 3 in the lungs (15%), 2 in the gastrointestinal and cervical lymph nodes, and 1 in the bones and pancreas(%5). CONCLUSION: 68Ga-DOTA-peptide PET/CT could find the primary or metastasis sites of NETs with good quality images. In general, this modality can enhance the management in patients with NETs.

Stereological study of the placental structure in abortion-prone mice model (CBA/J×DBA/2J).

Recurrent spontaneous abortion (RSA) is an important reproductive health issue defined as the loss of two or more consecutive pregnancies before the 20th week of gestation, affecting 2-5% of couples. This study aimed to evaluate the volume, number of cells, and length of the vessels in the placenta in normal and abortion-prone (AP) pregnant mice on gestational day (gd) 13.5. Fetal and placental tissues of female CBA/J mated DBA/2J (AP group) and BALB/c (normal pregnant group) were collected and prepared for stereological assessments on gd13.5. The volumes of the placenta and its main layers decidua basalis (Db), junctional zone (Jz), and labyrinth zone (Lz) were investigated. The number of spongiotrophoblast cells, glycogen cells, giant cells, trophoblast cells, lymphocytes, and neutrophils were estimated as well. The AP group showed a reduction in the volume of the placenta (48.7%) and its components. Moreover, the number of spongiotrophoblast cells (66.7%), glycogen cells (76.2%), giant cells (73.3%), and trophoblast cells (81.4%) was decreased in AP compared to normal pregnant (NP) mice. Also, in AP group recognized a 10-fold increase in the number of lymphocytes and a four-fold increase in the number of neutrophils in comparison to the NP group (p < 0.05). Activation of different immune cell types might induce systemic inflammation at the feto-maternal interface, resulting in impaired placenta formation and abortion.

Analysis of the frequency of type 2 innate lymphoid cells and regulatory T cells in abortion-prone mice.

Recurrent spontaneous abortion (RSA) is the most common pregnancy related complication, affecting 1-5 % of pregnancies. Despite hormonal, genetic and anatomical factors that result in abortion, impairment of immune response at the feto-maternal interface during the first trimester of pregnancy is also one of the main causes of RSA. In the present study, we evaluated the frequency of blood and uterine group 2 innate lymphoid cells (ILC2s), their subsets and regulatory T cells (Tregs) in CBA/J × DBA/2 J as an abortion-prone model compared to normal pregnant (NP) mice using immunophenotyping. Results indicated that the percentages of ILC2s were significantly decreased in the AP group compared to the NP group at mid-gestation (P ≤ 0.01). Moreover, the percentages of both blood and uterine nILC2s were increased in NP mice at mid-gestation (P ≤ 0.01, and P ≤ 0.05, respectively), while iILC2s significantly increased in AP mice at mid-gestation (P ≤ 0.01, and P ≤ 0.05, respectively). Tregs were reduced in AP mice at both early and mid-gestation stages (P ≤ 0.01). Overall, our findings suggest that the changes in blood and uterine ILC2s might be associated with abortion in mice.

Evaluation of IL-17 and IL-35 Serum Levels in Patients with Preeclampsia.

BACKGROUND: Pre-eclampsia (PE) is the most common pregnancy complication affecting 2-8% of all pregnancies. PE could lead to maternal and prenatal morbidity. Imbalanced cytokine network and altered levels of several inflammatory and anti-inflammatory cytokines have been reported in PE. Because of scare information regarding the roles of IL-17 and IL-35 in PE, the current study aimed to investigate the serum level of these cytokines in a group of Iranian women suffering from PE. METHODS: Serum samples were collected from 100 pre-eclamptic and 100 healthy pregnant women. Patients and controls were matched for age, ethnicity and body mass index. The level of IL-35 and IL-17 were evaluated by ELISA technique. T test and one-way ANOVA with Tukey Post-Hoc test were used for analysis and p<0.05 were assumed significant. RESULTS: The serum level of IL-35 was increased in pre-eclamptic subjects as compared with healthy pregnant women (p<0.001). There was no significant difference in the serum level of IL-17 between pre-eclamptic and healthy pregnant women (p=0.73). Moreover, the results of the present study also showed that the pregnant women with severe pre-eclampsia had higher level of IL-35 in their sera when compared to those with mild form of the disease (p<0.001). In addition, the serum level of IL-35 was significantly elevated in women with higher proteinuria (p<0.001). CONCLUSION: Based on the our results, it seems that elevated levels of IL-35 in sera of pre-eclamptic women might work as a marker to evaluate the severity of the preeclampsia.

Factors Affecting the Occurrence of Spinal Reflexes in Brain Dead Cases.

OBJECTIVES: Brain death is defined as the permanent absence of all cortical and brain stem reflexes. A wide range of spontaneous or reflex movements that are considered medullary reflexes are observed in heart beating cases that appear brain dead, which may create uncertainty about the diagnosis of brain death and cause delays in deceased-donor organ donation process. We determined the frequency and type of medullary reflexes and factors affecting their occurrence in brain dead cases. MATERIALS AND METHODS: During 1 year, 122 cases who fulfilled the criteria for brain death were admitted to the special intensive care unit for organ procurement of Masih Daneshvari Hospital. Presence of spinal reflexes was evaluated by trained coordinators and was recorded in a form in addition to other information including demographic characteristics, cause of brain death, time from detection of brain death, history of craniotomy, vital signs, serum electrolyte levels, and parameters of arterial blood gas determination. RESULTS: Most cases (63%) included in this study were male, and mean age was 33 ± 15 y. There was > 1 spinal reflex observed in 40 cases (33%). The most frequent reflex was plantar response (17%) following by myoclonus (10%), triple flexion reflex (9%), pronator extension reflex (8%), and undulating toe reflex (7%). Mean systolic blood pressure was significantly higher in cases who exhibited medullary reflexes than other cases (126 ± 19 mm Hg vs 116 ± 17 mm Hg; P = .007). CONCLUSIONS: Spinal reflexes occur frequently in brain dead cases, especially when they become hemodynamically stable after treatment in the organ procurement unit. Observing these movements by caregivers and family members has a negative effect on obtaining family consent and organ donation. Increasing awareness about spinal reflexes is necessary to avoid suspicion about the brain death diagnosis and delays in organ donation.

Role of laryngeal mask airway in interventional bronchoscopy procedures for upper tracheal stenosis: case series.

BACKGROUND: Bronchoscopic interventional procedures are novel means of treating airway lesions which are less invasive and well tolerated for patients with endo-luminal lesions, but managing the airway and oxygenating the patient in a field that is shared by both anesthesiologist and bronchoscopist is a major concern. Also in cases with subglottic and upper tracheal stenotic lesions an airway device placed inside the lumen interferes with the procedure and occasionally bears the hazard of ignition. Therefore, an airway device placing above the glottis with effective oxygenation is required. Laryngeal mask airway is a supra-glottic device which facilitates assisted or spontaneous positive pressure ventilation. METHODS: In this study, eight patients with subglottic stenoses due to different etiologies are presented who underwent fiberoptic bronchoscopy and therapeutic interventions through laryngeal mask. RESULTS: In all these patients, we experienced simple access to the vocal cord, glottis and trachea and also the lesion, besides effective oxygenation of the patient. Furthermore, bronchoscopist and patients were both comfortable with the procedures. CONCLUSION: Laryngeal mask airway could be regarded as a reliable alternative for airway management during interventional bronchoscopic procedures, especially when they are located near the glottis or in the upper third of the trachea.

Using Home Spirometry for Follow up of Lung Transplant Recipients: "A Pilot Study".

BACKGROUND: Lung transplantation is considered the ultimate treatment for some patients, but due to the specific condition of patients undergoing it, follow up is a major concern. The aim of this study was to evaluate the role of home spirometry in follow up of lung transplant recipients and early detection of complications in these patients. MATERIALS AND METHODS: A PC-based portable spirometry set was used to evaluate the well being of two lung transplant recipients on a regular daily basis for a 6-month period. Patient satisfaction and compliance, and device sensitivity in detecting complications were evaluated. Results of follow up were compared with 2 matched control patients. RESULTS: Patient adherence to home spirometry was 80% in one and 61% in the other patient and both patients were satisfied with the method, although this satisfaction declined towards the end of the study period. The main reason for low adherence was insufficient internet access. This method succeeded in early detection of infectious complications. CONCLUSION: Home spirometry seems to be a reliable method for follow up of lung transplant recipients, but further studies in a larger group of patients is recommended.