Transcript-specific enrichment enables profiling rare cell states via scRNA-seq.

Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing (scRNA-seq) identifies many rare populations of interest that express specific marker transcript combinations, traditional flow sorting limits our ability to enrich these populations for further profiling, including requiring cell surface markers with high-fidelity antibodies. Additionally, many single-cell studies require the isolation of nuclei from tissue, eliminating the ability to enrich learned rare cell states based on extranuclear protein markers. To address these limitations, we describe Programmable Enrichment via RNA Flow-FISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations from complex cellular mixtures defined by the presence or absence of specific RNA transcripts. Across immune populations (n = 141,227 cells) and fresh-frozen and formalin-fixed paraffin-embedded brain tissue (n = 29,522 nuclei), we demonstrate the sorting logic that can be used to enrich for cell populations via RNA-based cytometry followed by high-throughput scRNA-seq. Our approach provides a rational, programmable method for studying rare populations identified by one or more marker transcripts.

Late miscarriage and intraamniotic candidiasis in a woman with a retained intrauterine contraceptive device.

PIP: A case report is presented in which a 27-year-old patient, gravida 3, para 2, was admitted at 21 weeks of gestation with uterine contractions. She had had an IUD inserted 3 years before the present pregnancy, and an attempt to remove it when pregnancy was first diagnosed failed. An ultrasound examination at 7 weeks confirmed the gestational age and the presence of an IUD. On admission, ultrasound examination revealed a normal, live fetus with a normal amount of amniotic fluid. The patient was treated with sedation (Meperidine 100 mg and Phenergan 50 mg) and parenteral fluids. Several hours later the cervix was effaced and 4 cm dilated. Under ultrasound guidance, a transabdominal amniocentesis was performed. 20 ml of turbid, greenish amniotic fluid were obtained for Gram stain, white blood cell count, Limulus amebocyte lysate, aerobic, anaerobic and Mycoplasma cultures. Gram stain examination revealed the presence of yeasts. The white blood cell count of amniotic fluid revealed 9000 cells/ml with 73% lymphocytes and 17% polymorphonuclears. The Limulus amebocyte lysate test was negative. The membranes ruptured and a live male fetus weighing 430 g was delivered who died soon afterwards. The placenta was evacuated with the help of suction curettage. The retained IUD was removed with the placenta. Later, Candida was cultured from the amniotic fluid, and from the maternal and fetal sides of the placenta and the fetal skin. The patient was discharged after 5 days of antimycotic treatment was Ketoconazole. The placenta weighed 200 g. The chorionic plate and placental membranes showed histological signs of acute infection with multiple hyphae. The inflammatory infiltrate was composed of granulocytes and fungi. The lungs showed evidence of bronchopneumonia with an inflammatory infiltrate of granulocyte and macrophages. Early diagnosis of the cause of intraamniotic infection accords the best chance of success to antimycotic and other treatments in the prevention of fetal loss.^ieng

[Resistance in uterine arcuate arteries of pregnant hypertensive patients].

Hypertensive pregnant women were examined by continuous Doppler-ultrasound to measure resistance to blood flow in the uterine arcuate arteries. 20/26 had increased resistance. Those with bilateral increased resistance delivered after a mean of 33.2 weeks of pregnancy, 46.1% of them by cesarean section, and the mean birthweight was 1743 g. Those with unilateral increased resistance delivered after a mean of 38.0 weeks, 28.5% by cesarean section, and the mean birthweight was 2813 g. Measurement of blood flow by the Doppler method seems to be useful in the management of hypertensive disorders in pregnancy, as it helps to assess deficiency of the uterine circulation.