Intravital imaging via two-photon microscopy (TPM) is a useful tool for observing and delineating biological events at the cellular and molecular levels in live animals in a time-lapse manner. This imaging method provides spatiotemporal information with minimal phototoxicity while penetrating a considerable depth of intact organs in live animals.Although various organs can be visualized using intravital imaging, in the field of neuroscience, the brain is the main organ whose cell-to-cell interactions are imaged using this technique. Intravital imaging of brain disease in mouse models acts as an abundant source of novel findings for studying cerebral etiology. Neutrophil infiltration is a wellknown hallmark of inflammation; in particular, the crucial impact of neutrophils on the inflamed brain has frequently been reported in literature. Neutrophil extracellular traps (NETs) have drawn attention as an intriguing feature over the last couple of decades, opening a new era of research on their underlying mechanisms and biological effects.However, the actual role of NETs in the body is still controversial and is in parallel with a poor understanding of NETs in vivo. Although several experimental methods have been used to determine NET generation in vitro, some research groups have applied intravital imaging to detect NET formation in the inflamed organs of live mice. In this review, we summarize the advantages of intravital imaging via TPM that can also be used to characterize NET formation, especially in inflamed brains triggered by systemic inflammation. To study the function and migratory pattern of neutrophils, which is critical in triggering the innate immune response in the brain, intravital imaging via TPM can provide new perspectives to understand inflammation and the resolution process.

Neutrophil extracellular traps (NETs) exert a novel function of trapping pathogens. Released NETs can accumulate in inflamed tissues, be recognized by other immune cells for clearance, and lead to tissue toxicity. Therefore, the deleterious effect of NET is an etiological factor, causing several diseases directly or indirectly. NLR family pyrin domain containing 3 (NLRP3) in neutrophils is pivotal in signaling the innate immune response and is associated with several NET-related diseases. Despite these observations, the role of NLRP3 in NET formation in neuroinflammation remains elusive. Therefore, we aimed to explore NET formation promoted by NLRP3 in an LPS-induced inflamed brain. Wild-type and NLRP3 knockout mice were used to investigate the role of NLRP3 in NET formation. Brain inflammation was systemically induced by administering LPS. In such an environment, the NET formation was evaluated based on the expression of its characteristic indicators. DNA leakage and NET formation were analyzed in both mice through Western blot, flow cytometry, and in vitro live cell imaging as well as two-photon imaging. Our data revealed that NLRP3 promotes DNA leakage and facilitates NET formation accompanied by neutrophil death. Moreover, NLRP3 is not involved in neutrophil infiltration but is predisposed to boost NET formation, which is accompanied by neutrophil death in the LPS-induced inflamed brain. Furthermore, either NLRP3 deficiency or neutrophil depletion diminished pro-inflammatory cytokine, IL-1β, and alleviated blood-brain barrier damage. Overall, the results suggest that NLRP3 exacerbates NETosis in vitro and in the inflamed brain, aggravating neuroinflammation.These findings provide a clue that NLRP3 would be a potential therapeutic target to alleviate neuroinflammation.

Herlyn-Werner-Wunderlich syndrome is a rare congenital anomaly of the urogenital tract, which is characterized by the triad of uterus didelphys, obstructed hemivagina, and ipsilateral renal agenesis. It usually presents at puberty with pelvic pain, dysmenorrhea, and a vaginal or pelvic mass. Although rare, it may present with purulent vaginal discharge due to secondary infection of the obstructed hemivagina, making diagnosis difficult. A careful pelvic examination to identify the cervix and vagina is the key to the diagnosis of Müllerian duct anomalies and magnetic resonance imaging can provide additional useful information. The optimal treatment is full excision and marsupialization of the obstructing vaginal septum so that both uteri can drain through the patent vagina. The authors report a case of a 22-year-old female with an unusual presentation of Herlyn-Werner-Wunderlich syndrome complicated by pyocolpos, which was successfully managed by vaginal septum resection and drainage of pus.
Adolescent ; Cervix Uteri ; Coinfection ; Diagnosis ; Drainage ; Dysmenorrhea ; Female ; Gynecological Examination ; Humans ; Magnetic Resonance Imaging ; Pelvic Pain ; Puberty ; Suppuration ; Uterus ; Vagina ; Vaginal Discharge ; Young Adult

OBJECTIVE: The purpose of this study was to evaluate the group B streptococcus (GBS) colonization rate in pregnant Korean women using selective culture media for GBS and to identify obstetrical complications and GBS-induced early-onset neonatal sepsis. METHODS: We evaluated 1,014 pregnant women who delivered at Busan Paik Hospital between January 2015 and December 2016. GBS colonization was assessed using chromID Strepto B agar. We evaluated GBS colonization in pregnant women, as well as the obstetrical complication and GBS-induced neonatal sepsis rates. RESULTS: The total GBS colonization rate was 11.6% (117/1,014). No significant increase was observed in the rate of pregnancy-related complications between the GBS-positive and the GBS-negative groups. Among the 134 neonates born to colonized mothers, early neonatal sepsis was reported in 2 neonates (1.5%); however, these were cases of non-GBS-induced sepsis. CONCLUSION: The GBS colonization rate (using selective culture media) in this study involving pregnant Korean women showed a higher colonization rate than that previously reported in Korea. Therefore, based on this study, we recommend GBS screening and the administration of intrapartum antibiotic prophylaxis in pregnant Korean women.
Agar ; Antibiotic Prophylaxis ; Busan ; Colon* ; Culture Media ; Female ; Humans ; Infant, Newborn ; Korea* ; Mass Screening ; Mothers ; Pregnant Women* ; Prevalence* ; Sepsis ; Streptococcus agalactiae ; Streptococcus* ; Tertiary Care Centers* ; Tertiary Healthcare*

Adult ; Amenorrhea ; Congenital Abnormalities ; Drug Therapy ; Dysgerminoma ; Female ; Humans ; Karyotype ; Uterus