Efficacy and Safety of MED-01 Probiotics on Vaginal Health: A 12-Week, Multicenter, Randomized, Double-Blind, Placebo-Controlled Clinical Trial.

Bacterial vaginosis (BV) is the most common disease in women of childbearing age and is caused by the growth of abnormal microbiota in the vagina. Probiotic consumption can be an effective alternative treatment to preserve or improve vaginal health. In the present study, MED-01, a complex of five strains of probiotic candidates isolated from the vagina of Korean women, was used. This study was designed as a 12-week, randomized, multicenter, double-blind, placebo-controlled clinical trial to evaluate the efficacy and safety of MED-01 on vaginal health. A total of 101 reproductive-aged women with a Nugent score of 4-6 took MED-01 (5.0 × 109 CFU) or a placebo once a day, and 76 participants completed the procedure. MED-01 significantly reduced the Nugent score compared with the placebo. Quantitative PCR analysis confirmed that Lactobacillus plantarum was significantly increased in the vagina, whereas harmful bacteria such as Mobiluncus spp., Gardnerella vaginalis, and Atopobium vaginae were suppressed after 12 weeks of MED-01 ingestion. No adverse events to the test food supplements were observed in the participants. These results confirmed that MED-01 can be used as a probiotic for treating BV, as it improves the vaginal microbiota.

Effect of Topical Anesthesia on Pain from Needle Insertion and Injection and Its Relationship with Anxiety in Patients Awaiting Apical Surgery: A Randomized Double-blind Clinical Trial.

INTRODUCTION: We aimed to simultaneously investigate the effects of topical anesthesia on needle insertion and injection pain in the labial mucosa of the maxillary central incisors of patients awaiting apical surgery and to assess the relationship between patients' anxiety and pain scores. METHODS: Forty-four patients scheduled for apical surgery of the maxillary anterior incisor or canine were included, and all completed the Modified Dental Anxiety Scale (MDAS) questionnaire. One piece of Xylocaine (AstraZeneca, Sodertalje, Sweden) gauze was applied to the right or left side of the labial vestibule below the central incisor according to a randomization process, and 1 piece of water gauze was applied to the contralateral side of the labial vestibule. Each piece of gauze remained in place for 2 minutes. The subjects were asked to rate their pain according to the numeric rating scale immediately after needle insertion and anesthetic solution injection. RESULTS: Topical anesthetic application significantly reduced both insertion- and injection-related pain. Injection pain was significantly higher than insertion pain throughout the experiment. The difference in pain scores between the placebo and topical anesthetic groups was significantly greater for insertion pain than injection pain. The group with higher MDAS scores showed significantly higher pain scores, except for insertion pain reported by the topical anesthetic group, which did not show a significant difference between MDAS score groups. CONCLUSIONS: The topical anesthetic was highly effective for both insertion and injection pain during infiltration anesthesia in the maxillary central incisors. Highly anxious patients reported higher pain scores; however, topical anesthetics reduced the effect of anxiety on increasing pain.

Intrathecal Transplantation of Embryonic Stem Cell-Derived Spinal GABAergic Neural Precursor Cells Attenuates Neuropathic Pain in a Spinal Cord Injury Rat Model.

Neuropathic pain following spinal cord injury (SCI) is a devastating disease characterized by spontaneous pain such as hyperalgesia and allodynia. In this study, we investigated the therapeutic potential of ESC-derived spinal GABAergic neurons to treat neuropathic pain in a SCI rat model. Mouse embryonic stem cell-derived neural precursor cells (mESC-NPCs) were cultured in media supplemented with sonic hedgehog (SHH) and retinoic acid (RA) and efficiently differentiated into GABAergic neurons. Interestingly, low doses of SHH and RA induced MGE-like progenitors, which expressed low levels of DARPP32 and Nkx2.1 and high levels of Irx3 and Pax6. These cells subsequently generated the majority of the DARPP32(-) GABAergic neurons after in vitro differentiation. The spinal mESC-NPCs were intrathecally transplanted into the lesion area of the spinal cord around T10-T11 at 21 days after SCI. The engrafted spinal GABAergic neurons remarkably increased both the paw withdrawal threshold (PWT) below the level of the lesion and the vocalization threshold (VT) to the level of the lesion (T12, T11, and T10 vertebrae), which indicates attenuation of chronic neuropathic pain by the spinal GABAergic neurons. The transplanted cells were positive for GABA antibody staining in the injured region, and cells migrated to the injured spinal site and survived for more than 7 weeks in L4-L5. The mESC-NPC-derived spinal GABAergic neurons dramatically attenuated the chronic neuropathic pain following SCI, suggesting that the spinal GABAergic mESC-NPCs cultured with low doses of SHH and RA could be alternative cell sources for treatment of SCI neuropathic pain by stem cell-based therapies.

Synchronous activation of gonadotropin-releasing hormone gene transcription and secretion by pulsatile kisspeptin stimulation.

Pulsatile release of hypothalamic gonadotropin-releasing hormone (GnRH) is essential for pituitary gonadotrope function. Although the importance of pulsatile GnRH secretion has been recognized for several decades, the mechanisms underlying GnRH pulse generation in hypothalamic neural networks remain elusive. Here, we demonstrate the ultradian rhythm of GnRH gene transcription in single GnRH neurons using cultured hypothalamic slices prepared from transgenic mice expressing a GnRH promoter-driven destabilized luciferase reporter. Although GnRH promoter activity in each GnRH neuron exhibited an ultradian pattern of oscillations with a period of ∼10 h, GnRH neuronal cultures exhibited partially synchronized bursts of GnRH transcriptional activity at ∼2-h intervals. Surprisingly, pulsatile administration of kisspeptin, a potent GnRH secretagogue, evoked dramatic synchronous activation of GnRH gene transcription with robust stimulation of pulsatile GnRH secretion. We also addressed the issue of hierarchical interaction between the circadian and ultradian rhythms by using Bmal1-deficient mice with defective circadian clocks. The circadian molecular oscillator barely affected basal ultradian oscillation of GnRH transcription but was heavily involved in kisspeptin-evoked responses of GnRH neurons. In conclusion, we have clearly shown synchronous bursts of GnRH gene transcription in the hypothalamic GnRH neuronal population in association with episodic neurohormone secretion, thereby providing insight into GnRH pulse generation.

Acupuncture decreases ischemia-induced apoptosis and cell proliferation in dentate gyrus of gerbils.

BACKGROUND: Acupuncture has been used for the enhancement of functional recovery from various disorders. In the present study, the effect of acupuncture on the apoptosis and new cell proliferation in the hippocampal dentate gyrus of gerbils (n = 25) following transient global ischemia was investigated. METHODS: To determine the level of apoptosis and cell proliferation, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) were employed respectively. RESULTS: In the dentate gyrus of ischemic gerbils, the number of both TUNEL- and BrdU-positive cells (66.01 +/- 2.45/mm(2) and 514.38 +/- 44.90/mm(2)) was significantly increased compared to that of the sham-operated gerbils (11.25 +/- 1.85/mm(2) and 111.47 +/- 10.95/mm(2)). Among the acupuncture (ST36, LI4 or non-acupoint) treated groups, ST36 acupoint treated group showed the most potent apoptosis (20.52 +/- 2.40/mm(2)) and proliferation (159.38 +/- 24.05/mm(2)) suppressive effects ( approximately 70% decreases in both apoptosis and cell proliferation). CONCLUSION: These results may suggest that acupuncture treatment alleviates ischemia-induced apoptosis and presents possible therapeutic potentials in the recovery from ischemic cerebral injury.

Cooperative actions of Tra2alpha with 9G8 and SRp30c in the RNA splicing of the gonadotropin-releasing hormone gene transcript.

In earlier studies, we demonstrated that excision of the first intron (intron A) from the gonadotropin-releasing hormone (GnRH) transcript is highly cell type- and developmental stage-specific. The removal of GnRH intron A requires exonic splicing enhancers on exons 3 and 4 (ESE3 and ESE4, respectively). Tra2alpha,a serine/arginine-rich (SR)-like protein, specifically binds to ESE4, although it requires additional nuclear co-factors for efficient removal of this intron. In the present study, we demonstrate the cooperative action of multiple SR proteins in the regulation of GnRH pre-mRNA splicing. SRp30c specifically binds to both ESE3 and ESE4, whereas 9G8 binds to an element in exon 3 and strongly enhances the excision of GnRH intron A in the presence of minimal amount of other nuclear components. Interestingly, Tra2alpha can interact with either 9G8 or SRp30c, whereas no interaction between 9G8 and SRp30c is observed. Tra2alpha has an additive effect on the RNA binding of these proteins. Overexpression or knock-down of these three proteins in cultured cells further suggests their essential role in intron A excision activities, and their presence in GnRH neurons of the mouse preoptic area further strengthens this possibility. Together, these results indicate that interaction of Tra2alpha with 9G8 and SRp30c appears to be crucial for ESE-dependent GnRH pre-mRNA splicing, allowing efficient generation of mature mRNA in GnRH-producing cells.