Hindbrain Adenosine 5-Triphosphate (ATP)-Purinergic Signaling Triggers LH Surge and Ovulation via Activation of AVPV Kisspeptin Neurons in Rats.

Ovulation disorders are a serious problem for humans and livestock. In female rodents, kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) are responsible for generating a luteinizing hormone (LH) surge and consequent ovulation. Here, we report that adenosine 5-triphosphate (ATP), a purinergic receptor ligand, is a possible neurotransmitter that stimulates AVPV kisspeptin neurons to induce an LH surge and consequent ovulation in rodents. Administration of an ATP receptor antagonist (PPADS) into the AVPV blocked the LH surge in ovariectomized (OVX) rats treated with a proestrous level of estrogen (OVX + high E2) and significantly reduced the ovulation rate in proestrous ovary-intact rats. AVPV ATP administration induced a surge-like LH increase in OVX + high E2 rats in the morning. Importantly, AVPV ATP administration could not induce the LH increase in Kiss1 KO rats. Furthermore, ATP significantly increased intracellular Ca2+ levels in immortalized kisspeptin neuronal cell line, and coadministration of PPADS blocked the ATP-induced Ca2+ increase. Histologic analysis revealed that the proestrous level of estrogen significantly increased the number of P2X2 receptor (an ATP receptor)-immunopositive AVPV kisspeptin neurons visualized by tdTomato in Kiss1-tdTomato rats. The proestrous level of estrogen significantly increased varicosity-like vesicular nucleotide transporter (a purinergic marker)-immunopositive fibers projecting to the vicinity of AVPV kisspeptin neurons. Furthermore, we found that some hindbrain vesicular nucleotide transporter-positive neurons projected to the AVPV and expressed estrogen receptor α, and the neurons were activated by the high E2 treatment. These results suggest that hindbrain ATP-purinergic signaling triggers ovulation via activation of AVPV kisspeptin neurons.SIGNIFICANCE STATEMENT Ovulation disorders, which cause infertility and low pregnancy rates, are a serious problem for humans and livestock. The present study provides evidence that adenosine 5-triphosphate, acting as a neurotransmitter in the brain, stimulates kisspeptin neurons in the anteroventral periventricular nucleus, known as the gonadotropin-releasing hormone surge generator, via purinergic receptors to induce the gonadotropin-releasing hormone/luteinizing hormone surge and ovulation in rats. In addition, histologic analyses indicate that adenosine 5-triphosphate is likely to be originated from the purinergic neurons in the A1 and A2 of the hindbrain. These findings may contribute to new therapeutic controls for hypothalamic ovulation disorders in humans and livestock.

The Effect of Static Stretching Duration on Muscle Blood Volume and Oxygenation.

ABSTRACT: Matsuo, H, Kubota, M, Shimada, S, Kitade, I, Matsumura, M, Nonoyama, T, Koie, Y, Naruse, H, Takahashi, A, Oki, H, Kokubo, Y, and Matsumine, A. The effect of static stretching duration on muscle blood volume and oxygenation. J Strength Cond Res 36(2): 379-385, 2022-Muscle blood volume increases due to stretching; however, the minimum duration of stretching to sustainably increase the muscle blood volume after stretching has not yet been elucidated. This study examined whether the duration of static stretching influenced the muscle blood volume and oxygenation. Ten healthy male subjects participated in this controlled laboratory study. Static stretching of the gastrocnemius muscle was performed for 5 durations (20 seconds, and 1, 2, 5, and 10 minutes). Changes in both the total-Hb (ΔtHb), as an index of blood volume, and tissue oxygenation index (ΔTOI) from baseline were determined using near-infrared spectroscopy. Both the ΔtHb and ΔTOI decreased during stretching and increased after stretching. The minimum value of ΔtHb during stretching did not differ in each of the 5 durations, but minimum ΔTOI progressively decreased with longer durations of stretching. The peak value of ΔtHb after stretching increased with longer durations of stretching. The value of ΔtHb at 5 minutes after the end of stretching increased with more than 2 minutes of stretching compared with 20 seconds of stretching, although the value of ΔtHb did not significantly differ between the 2, 5, and 10 minutes' durations. These findings suggest that a longer duration of stretching elicits a decrease in muscle oxygenation during stretching, and an increase in both the muscle blood volume and oxygenation after stretching. The results indicated that the minimum duration of stretching to sustain an increase in the muscle blood volume after stretching is 2 minutes.

Intraarticular injection of processed lipoaspirate cells has anti-inflammatory and analgesic effects but does not improve degenerative changes in murine monoiodoacetate-induced osteoarthritis.

BACKGROUND: Previous basic research and clinical studies examined the effects of mesenchymal stem cells (MSCs) on regeneration and maintenance of articular cartilage. However, our pilot study suggested that MSCs are more effective at suppressing inflammation and pain rather than promoting cartilage regeneration in osteoarthritis. Adipose tissue is considered a useful source of MSCs; it can be harvested easily in larger quantities compared with the bone marrow. The present study was designed to evaluate the anti-inflammatory, analgesic, and regenerative effects of intra-articularly injected processed lipoaspirate (PLA) cells (containing adipose-derived MSCs) on degenerative cartilage in a rat osteoarthritis model. METHODS: PLA cells were isolated from subcutaneous adipose tissue of 12-week-old female Sprague-Dawley rats. Osteoarthritis was induced by injection of monoiodoacetate (MIA). Each rat received 1 × 106 MSCs into the joint at day 7 (early injection group) and day 14 (late injection group) post-MIA injection. At 7, 14, 21 days after MIA administration, pain was assessed by immunostaining and western blotting of dorsal root ganglion (DRG). Cartilage quality was assessed macroscopically and by safranin-O and H&E staining, and joint inflammation was assessed by western blotting of the synovium. RESULTS: The early injection group showed less cartilage degradation, whereas the late injection group showed cartilage damage similar to untreated OA group. The relative expression level of CGRP protein in DRG neurons was significantly lower in the two treatment groups, compared with the untreated group. CONCLUSIONS: Intra-articular injection of PLA cells prevented degenerative changes in the early injection group, but had little effect in promoting cartilage repair in the late injection group. Interestingly, intra-articular injection of PLA cells resulted in suppression of inflammation and pain in both OA groups. Further studies are needed to determine the long-term effects of intra-articular injection of PLA cells in osteoarthritis.

Prognostic factors and optimal management for patients with cervical spinal cord injury without major bone injury.

BACKGROUND: Even though the number of patients with cervical spinal cord injury (CSCI) without major bone injury is increased, the treatment with either surgery or conservative measures remains controversial. The aim of this study was to assess its prognostic value in the prediction of useful motor recovery and to clarify whether the patients should be treated surgically are present. METHODS: We reviewed 63 patients (conservative, n = 36; surgery, n = 27) with CSCI without major bone injury (Frankel A-C). Neurological examination using modified Frankel grade at admission and 6 months after injury and International Stoke Mandeville Games (ISMG) classification at subacute phase after injury, MRI findings including rate of spinal cord compression, extent of cord damage and type of signal intensity change were assessed. RESULTS: Thirty-five of 63 patients were improved to walk at 6 months after injury. In multivariate analysis, rate of spinal cord compression, extent of cord damage and improvement of ISMG grade were associated with useful motor recovery. There was no difference in the neurological improvement between conservative and surgical groups. However, patients with spinal cord compression of ≥33.2% showed better motor recovery at 6 months post-injury after surgery than those treated conservatively. There was a positive correlation between the improvement of ISMG grade at subacute phase and Frankel grade at 6 months post-injury. It is difficult to obtain satisfactory surgical outcome for patients with Frankel A or B1 on admission and/or extensive spinal cord damage on T2-weighted image. CONCLUSIONS: Conservative treatment is recommended for patients with CSCI without major bone injury. However, we also recommend surgical treatment to acquire walking ability for patients with spinal cord compression of ≥33.2% and low ISMG grade at subacute phase. Among such patients, careful consideration should be given to patients with Frankel A or B1 and/or extensive spinal cord damage on MRI.

Brain pericytes serve as microglia-generating multipotent vascular stem cells following ischemic stroke.

BACKGROUND: Microglia are the resident macrophage population of the central nervous system (CNS) and play essential roles, particularly in inflammation-mediated pathological conditions such as ischemic stroke. Increasing evidence shows that the population of vascular cells located around the blood vessels, rather than circulating cells, harbor stem cells and that these resident vascular stem cells (VSCs) are the likely source of some microglia. However, the precise traits and origins of these cells under pathological CNS conditions remain unclear. METHODS: In this study, we used a mouse model of cerebral infarction to investigate whether reactive pericytes (PCs) acquire microglia-producing VSC activity following ischemia. RESULTS: We demonstrated the localization of ionized calcium-binding adaptor molecule 1 (Iba1)-expressing microglia to perivascular regions within ischemic areas. These cells expressed platelet-derived growth factor receptor-ß (PDGFRß), a hallmark of vascular PCs. PDGFRß(+) PCs isolated from ischemic, but not non-ischemic, areas expressed stem/undifferentiated cell markers and subsequently differentiated into various cell types, including microglia-like cells with phagocytic capacity. CONCLUSIONS: The study results suggest that vascular PCs acquire multipotent VSC activity under pathological conditions and may thus be a novel source of microglia.

[Curative Left Pulmonary Resection Combined with Total En Bloc Spondylectomy for Lung Cancer Invading the Second and Third Thoracic Vertebral Bodies].

Introduction of spinal surgery into the operation of lung cancer has made extensive surgical treatment feasible with acceptable long-term survival. We report our experience on total en bloc total spondylectomy for lung cancer invading the spine. A 60-year-old man was diagnosed with lung adenocarcinoma of the apicodorsal segment of the left lobe with invasion of the 2nd and 3rd thoracic vertebral bodies. After induction chemoradiotherapy, we performed en bloc resection through a posterolateral thoracotomy in the right decubitus position and a posterior median approach in the prone position. The thoracic manipulation was done earlier, making it useful for the dissection of the prevertebral plane from the posterior mediastinum at the upper thoracic level in addition to confirmation of non-N2 disease. Vertebral stabilization was achieved with rod fixation and placement of a titanium mesh cage between the remaining vertebral bodies.

[Total Gastrectomy for Advanced Gastric Cancer after Coronary Artery Bypass Grafting Using the Right Gastroepiploic Artery: Report of a Case].

An 82-year-old man, who had undergone coronary artery bypass grafting(CABG) with the right gastroepiploic artery( RGEA) 13 years previously, suffered with abdominal pain after meal and tarry stools, and was diagnosed with advanced gastric cancer. Gastroscopy revealed an advanced Borrmann type 4 cancer at the lesser curvature of the gastric body to the pyloric ring. The gastrogram showed poor extension and stenosis at the same part. Abdominal computed tomography showed the tumor reached the subserosal layer and infrapyloric lymph nodes were swollen. Abdominal angiography showed the RGEA graft remained well patent. Total gastrectomy with D2 lymph nodes dissection and arterial reconstruction between the splenic artery and the RGEA graft was performed. He has been well without any sign of cancer recurrence since the operation. Recently, more patients with CABG using RGEA are found to have gastric cancer and require the resection of RGEA for lymph nodes dissection as this case. We consider this procedure one of the options for advanced gastric cancer after coronary bypass grafting using RGEA.

[Chest Wall Reconstruction Using Titanium Plates Sandwiched Between Sheets after Resection of Chest Wall Chondrosarcoma].

Extensive chest wall resection carries the risk of difficult reconstruction and surgical complications. We report our experience on chest wall reconstruction using titanium plates for a wide thoracic defect after tumor resection. A 74-year-old man was diagnosed with chondrosarcoma of the 6th rib on the right. He needed extensive chest wall resection because of skip lesions on 4th rib noted on operative inspection, leaving a defect measuring 33 × 20 cm. Reconstruction using 5 transverse titanium plates sandwiched between an expanded polytetrafluoroethylene patch and a polypropylene mesh sheet stabilized the chest wall. This reconstruction allowed successful separation from ventilatory support after operation. The postoperative course was uneventful, and he was discharged on postoperative day 20. The advantages of this form of reconstruction over conventional prostheses are rigidity, and stability and usability.

[Perioperative Effect of Intraoperative Fluid Restriction with the Same Fixed Volume of 5 ml · kg(-1) · h(-1) in Patients Undergoing Major Abdominal Versus Thoracic Surgery].

BACKGROUND: Major abdominal surgery accompanies the higher magnitude of physiological stress response and may require an additional replacement fluid for the redistributed volume. Intraoperative volume restriction strategy is recommended to avoid fluid overload leading to increased mortality. We conducted a comparative study of the perioperative effects of intraoperative fluid restriction in abdominal versus thoracic surgery. METHODS: Each 15 patients having major abdominal or thoracic surgery were studied prospectively. All participants were identically given intraoperative iv crystalloid of 5 ml · kg(-1) · hr(-1) under combined epidural/general anesthesia. Plasma level of AVP, aldosterone, angiotensin II and IL-6 as well as body water composition by bioelectrical impedance analysis was examined at preoperative period, at the end of surgery and on the first postoperative day. RESULTS: In abdominal surgery group there was significantly less intraoperative urine output compared with thoracic surgery group. No significant differences were found between two groups in extracellular water volume chnages, AVP, aldosterone angiotensin II, IL-6 level and postoperative renal function. CONCLUSIONS: Restrictive fluid therapy with intraoperative crystalloid of 5 ml · kg(-1) · hr(-1) can be safely used with no serious adverse events in abdominal surgery. In conclusion we had better not make any traditional difference in intraoperative fluid management between abdominal and thoracic surgery even if their stress response differs in magnitude.

[Endoscopic Long Saphenous Vein Harvest from Femoralsite to Below the Knee through a Single Small Incision for Minimally Invasive Coronary Artery Bypass Grafting].

BACKGROUND: Endoscopic saphenous vein harvest( EVH) for coronary artery bypass grafting( CABG) has been proved to be effective in reduced wound complications without compromising graft patency and mortality. Although EVH was usually performed from femoral site, we performed endoscopic long saphenous vein harvest( ELSH) from femoral to below the knee through a single small incision. There were few reports in EVH from below the knee and ELSH. PATIENTS AND METHODS: Between September 2012 and June 2014, 13 patients who underwent ELSH in CABG. We assessed the initial evaluation in ELSH using the VirtuoSaph system. ELSH was performed by our unified procedure protocol to harvest good saphenous vein graft. RESULTS: The success rate of ELSH was 100% (13/13). The harvest time was 83.6±15.2 min, graft length was 52.1±5.6 cm, and number of side branches requiring repair was 2.6±1.6. Beating CABG was performed in all patients. The number of distal anastomoses was 3.8±0.8. Sequential bypass was 12 in total 23 saphenous vein grafts. The early patency was 95% (18/19 grafts) as evaluated by postoperative angiography or coronary enhanced computed tomography. All patients except 1( hospital death)were discharged without major or wound complications. CONCLUSIONS: The acceptable results and outcomes were demonstrated by our ELSH procedure protocol. ELSH is considered to be a feasible procedure to harvest enough graft length for 2 CABG and keep the other side of saphenous vein graft intact.

[Postoperative pain management in video-assisted thoracic surgery using a continuous unilateral intercostal analgesia].

BACKGROUND: Optimal pain management after video-assisted thoracic surgery (VATS) remains an open issue. We prospectively studied the analgesic effect of intercostal analgegia (ICA) by comparison with epidural analgesia. METHODS: Twenty-two patients undergoing VATS procedures were randomly divided into ICA (n = 8) or epidural (n = 14) group. Postoperatively 2 ml x hr(-1) of 0.2% ropivacaine was delivered continuously through intercostal or epidural catheter. Moreover, each group received the equal dose of fentanyl (0.25 x µg(-1). kg(-1) x hr(-1)) intravenously or epidurally. When no pain relief was achieved, iv fentanyl was given as a rescue. Requirement of additional fentanyl and pain score using a visual analogue scale (VAS) were documented for 19 hours. RESULTS: The mean pain scores at rest, mobilization and with coghing were slightly higher in the ICA group. Total additional dose of iv fentanyl was significantly different between the groups (ICA 147 ± 41 vs Epidural 39 ± 15 µg; P = 0.015). Pain scores and fentanyl requirements spread over the lower range. The mean of VAS in ICA group was less than 5 even at coughing, suggesting clinically irrelevant. CONCLUSIONS: In patients with coagulopathy, multimodal approach using intercostal analgesia supplemented by intravenous patient-controlled analgesia may be an alternative to epidural analgesia for postoperative pain management.

[Left ventricular outflow tract obstruction after mitral valve plasty; report of a case].

A 65-year-old female was admitted to our hospital with mitral regurgitation (MR). Transthoracic echocardiography showed severe mitral valve prolapse and subaortic septal hypertrophy with no pressure gradient. Mitral valve plasty consisted of artificial chorda implantation and ring annuloplasty was performed. During intensive care unit( ICU) stay after operation, systolic murmur and low cardiac output syndrome were noted and echocardiography revealed left ventricular outflow tract obstruction (LVOTO) without systolic anterior motion and MR. Cessation of catecholamine, volume administration, beta-blocker and negative inotropic drug like cibenzoline rapidly reduced LVOTO and the hemodynamic condition was improved. Even in a case of subaortic septal hypertrophy with no pressure gradient, emergence of LVOTO should be considered when new systolic murmur and low cardiac output syndrome appeared.

[Aortopulmonary fistula caused by aortic arch aneurysm rupture presenting severe left to right shunt and congestive heart failure].

A 74-year-old female was admitted to our hospital due to hyperkinetic congestive heart failure. Noninvasive positive pressure ventilation, continuous hemodialysis and inotropic support were needed preoperatively. Computed tomography revealed saccular aortic arch aneurysm with aortopulmonary fistula toward the left pulmonary artery. Cardiac catheterization revealed severe left to right shunt with the pulmonary flow/systemic flow (Qp/Qs) ratio of 6.2.Emergency operation was performed under hypothermic circulatory arrest followed by selective cerebral perfusion and lower body perfusion. The aortopulmonary fistula was directly closed, reinforced with the aneurysmal wall, and total arch replacement was performed. Although the patient temporarily recovered by endoscopic suction of the blood from the lungs and continuous hemodialysis filtration, she died because of multiple organ failure due to mesenteric ischemia on postoperative day 34.

Management of Cervical Spinal Cord Injury without Major Bone Injury in Adults.

The incidence of cervical spinal cord injury (CSCI) without major bone injury is increasing, possibly because older people typically have pre-existing cervical spinal canal stenosis. The demographics, neurological injury, treatment, and prognosis of this type of CSCI differ from those of CSCI with bone or central cord injury. Spine surgeons worldwide are debating on the optimal management of CSCI without major bone injury. Therefore, this narrative review aimed to address unresolved clinical questions related to CSCI without major bone injury and discuss treatment strategies based on current findings. The greatest divide among spine surgeons worldwide hinges on whether surgery is necessary for patients with CSCI without major bone injury. Certain studies have recommended early surgery within 24 h after injury; however, evidence regarding its superiority over conservative treatment remains limited. Delayed MRI may be beneficial; nevertheless, reliable factors and imaging findings that predict functional prognosis during the acute phase and ascertain the necessity of surgery should be identified to determine whether surgery/early surgery is better than conservative therapy/delayed surgery. Quality-of-life assessments, including neuropathic pain, spasticity, manual dexterity, and motor function, should be performed to examine the superiority of surgery/early surgery to conservative therapy/delayed surgery.

Improvement After Surgery in a Patient With Prolonged Tetraplegia Due to Cervical Spinal Cord Injury Without Bone Injury.

Treatment strategies for patients with cervical spinal cord injury (CSCI) without major bone injury in the acute phase are under debate. For CSCI without major bone injury, conservative treatment is often the first choice owing to the absence of fractures and spinal column instability. However, treatment of CSCI without major bone injury by either surgery or conservative measures remains controversial. We described a case of a 48-year-old man with cervical American Spinal Cord Injury Association Impairment Scale (AIS) grade C tetraplegia as a result of a fall. Computed tomography scan and magnetic resonance imaging revealed no fractures and widespread T2-hyperintense signal changes in the cord centered on C3-4. The paralyzed condition of his lower extremities remained unchanged with conservative treatment for eight months after the injury. Therefore, he underwent decompression surgery eight months after the injury. At two weeks postoperatively, he could transfer and walk using a walker. After discharge, he underwent regular home-visit rehabilitation and gradually improved his physical functions, including gait ability one year postoperatively. We encountered a case in which surgery and intensive rehabilitation eight months after the injury improved motor function. The combination of surgery in the chronic phase and postoperative rehabilitation can therefore improve the outcomes. The message in this paper is by no means a recommendation for "late surgery." However, we suggested that surgical treatment might be an option if the functional improvement is poor, as even quite late surgery can provide functional improvement.

Adipose-Derived Mesenchymal Stromal Cell Transplantation for Severe Spinal Cord Injury: Functional Improvement Supported by Angiogenesis and Neuroprotection.

Mesenchymal stromal cell transplantation alone is insufficient when motor dysfunction is severe; combination therapy with rehabilitation could improve motor function. Here, we aimed to analyze the characteristics of adipose-derived MSCs (AD-MSCs) and determine their effectiveness in severe spinal cord injury (SCI) treatment. A severe SCI model was created and motor function were compared. The rats were divided into AD-MSC-transplanted treadmill exercise-combined (AD-Ex), AD-MSC-transplanted non-exercise (AD-noEx), PBS-injected exercise (PBS-Ex), and no PBS-injected exercise (PBS-noEx) groups. In cultured cell experiments, AD-MSCs were subjected to oxidative stress, and the effects on the extracellular secretion of AD-MSCs were investigated using multiplex flow cytometry. We assessed angiogenesis and macrophage accumulation in the acute phase. Spinal cavity or scar size and axonal preservation were assessed histologically in the subacute phase. Significant motor function improvement was observed in the AD-Ex group. Vascular endothelial growth factor and C-C motif chemokine 2 expression in AD-MSC culture supernatants increased under oxidative stress. Enhanced angiogenesis and decreased macrophage accumulation were observed at 2 weeks post-transplantation, whereas spinal cord cavity or scar size and axonal preservation were observed at 4 weeks. Overall, AD-MSC transplantation combined with treadmill exercise training improved motor function in severe SCI. AD-MSC transplantation promoted angiogenesis and neuroprotection.

Imaging and Clinical Findings after Auto-mobilization in a Cervical Disk Herniation Patient with Prolonged Neck Pain: A 7-year Follow-up Case Report.

Background: Auto-mobilization (AM) is a treatment method that patients can use by themselves for pain relief. We report the case of a patient diagnosed with cervical disk herniation (CDH), with frequent recurrences of upper limb numbness and neck pain. The patient experienced a favorable outcome after cervical spine AM, as evidenced by the immediate and long-term relief of his symptoms as well as changes observed through imaging. Case: A 33-year-old-man diagnosed with CDH presented with frequent recurrences of upper limb numbness and neck pain. Radiographic and T2-weighted magnetic resonance imaging scans revealed cervical spine kyphosis and a left paracentral to intraforaminal lesion with disk herniation of protrusion type at C4-5. He was started on AM to elicit physiological lordosis of the cervical spine. This treatment was painless and did not cause withdrawal on discontinuation. AM improved the mobility of his cervical lower facet joints, reduced dysesthesia, and eliminated pain. Improvement in neck pain and cervical kyphosis and reduction of disk herniation were observed 2 years after initiating the intervention. Discussion: Appropriate physical therapy evaluation and cervical AM for this patient resulted in symptomatic relief and indirect disk herniation regression. By adding imaging findings to clinical findings, the effect of AM could be visualized, and the reliability of the therapeutic effect was further enhanced.