Interventional Challenges in Non-Tubal Ectopic Pregnancy.

Objective: Non-tubal ectopic pregnancies (EPs) are rare and potentially life threatening. The number is rising due to various risk factors and there are no uniform guidelines in the management of EPs. This study was done to assess risk factors and challenges in the management of EPs. Materials and methods: This is a retrospective observational descriptive study that was done at SDM College of Medical Sciences & Hospital, Shri Dharmasthala Manjunatheshwara University Dharwad, Karnataka India. Data was collected from the medical records section of all the patients of non-tubal ectopic pregnancies managed in our hospital from January 2020 to June 2021. The collected data were analyzed for demographic characteristics, risk factors and management. Results: The incidence of ectopic pregnancies in our institute was 6-7 per 1000 pregnancies, of which 20% of the ectopic pregnancies were non-tubal. The incidence was higher than the other studies, which could be due to our center being a tertiary care referral center. Cesarean scar ectopic pregnancies were the most common accounting for 60% of cases. The management varied from conservative to minimally invasive surgery to hysterectomy hysterectomy with bilateral internal iliac artery ligation, depending upon the clinical presentation, duration of gestation, presence of fetal cardiac activity and hemodynamic stability. The other non-tubal ectopic pregnancies were cervical, ovarian, corneal and heterotopic. Cervical pregnancy beyond 12 weeks of gestation was rare which was managed by conserving the uterus. Conclusion: Non-tubal ectopic pregnancies are rare. Early diagnosis requires a high index of suspicion if missed can lead to an array of complications leading to loss of fertility, morbidity, and mortality. The key step to avert the complications is early diagnosis and individualized treatment.

Generation of a recombinant version of a biologically active cell-permeant human HAND2 transcription factor from E. coli.

Transcription factor HAND2 has a significant role in vascularization, angiogenesis, and cardiac neural crest development. It is one of the key cardiac factors crucial for the enhanced derivation of functional and mature myocytes from non-myocyte cells. Here, we report the generation of the recombinant human HAND2 fusion protein from the heterologous system. First, we cloned the full-length human HAND2 gene (only protein-coding sequence) after codon optimization along with the fusion tags (for cell penetration, nuclear translocation, and affinity purification) into the expression vector. We then transformed and expressed it in Escherichia coli strain, BL21(DE3). Next, the effect (in terms of expression) of tagging fusion tags with this recombinant protein at two different terminals was also investigated. Using affinity chromatography, we established the one-step homogeneous purification of recombinant human HAND2 fusion protein; and through circular dichroism spectroscopy, we established that this purified protein had retained its secondary structure. We then showed that this purified human protein could transduce the human cells and translocate to its nucleus. The generated recombinant HAND2 fusion protein showed angiogenic potential in the ex vivo chicken embryo model. Following transduction in MEF2C overexpressing cardiomyoblast cells, this purified recombinant protein synergistically activated the α-MHC promoter and induced GFP expression in the α-MHC-eGFP reporter assay. Prospectively, the purified bioactive recombinant HAND2 protein can potentially be a safe and effective molecular tool in the direct cardiac reprogramming process and other biological applications.