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PubMed1991

Neonatal seminal vesicle mesenchyme induces a new morphological and functional phenotype in the epithelia of adult ureter and ductus deferens.

Mesenchyme from neonatal mouse and rat seminal vesicles (SVM) was grown in association with postnatal (adult) epithelial cells from the ureter (URE) and ductus deferens (DDE) in chimeric tissue recombinants composed of mouse mesenchyme and rat epithelium or vice versa. Functional cytodifferentiation was examined in these SVM + URE and SVM + DDE tissue recombinants with antibodies against major androgen-dependent seminal-vesicle-specific secretory proteins. Adult DDE and URE were induced to express seminal cytodifferentiation and produced the complete spectrum of major seminal vesicle secretory (SVS) proteins. The SVS proteins produced were specific for the species that provided the epithelium. In the case of SVM + URE recombinants, the URE, which normally lacks androgen receptors (AR), expressed AR. These results demonstrate that adult epithelial cells retain a developmental plasticity equivalent to their undifferentiated fetal counterparts and are capable of being reprogrammed to express a completely new morphological, biochemical and functional phenotype.

Animals↗
PubMed2000

Comparison of cardiac stroke volume measurement determined using stereological analysis of breath-hold cine MRI and phase contrast velocity mapping.

Cine MRI of the heart using segmented k-space pulse sequences permits multiphase images to be acquired in a single breath-hold. Whilst image quality is improved compared with conventional (non-segmented) cine imaging, subsequent analysis can be relatively time consuming. In this study, multiple slice breath-hold cine imaging of the heart was performed in 11 normal volunteers. Left and right ventricular stroke volume (SV) was estimated from the images by application of the Cavalieri method of modern design stereology in combination with point counting. The measured SVs were compared with those obtained by cine phase contrast velocity mapping in the ascending aorta and main pulmonary artery. Excellent agreement was found between the SVs determined by the two techniques, with mean differences (+/- one SD) of 0.41 +/- 3.00 ml and 0.41 +/- 4.78 ml for left and right ventricles, respectively. Comparison of left and right ventricular SV using stereology yielded a mean difference of 0.84 +/- 5.70 ml. Breath-hold data acquisition together with stereological analysis is demonstrated to be an accurate and unbiased technique for the rapid assessment of cardiac function.

Adult↗
PubMed1990

cDNA cloning of androgen-stimulated mRNAs in rat seminal vesicles: partial characterization of newly isolated cDNA clones, pSv-1 and pSv-2.

As the first step in surveying the molecular mechanism of androgen-responsive gene expression in rat seminal vesicles, the effect of androgen on the mRNAs was examined by in vitro translation assay. When the in vitro translation products of mRNAs from castrated animals (48 h) were compared with those from castrated and testosterone-treated animals (48 h) by SDS-PAGE, several discrete bands which were stimulated or repressed in response to androgen were observed in addition to major peptide bands of SVS IV and SVS V. From these findings, we constructed a partial cDNA library from the seminal vesicle poly(A +)RNAs of androgen-treated rats and screened by differential colony hybridization. Two distinct cDNA clones, pSv-1 and pSv-2, whose mRNAs were differentially stimulated in response to androgen and seemed to be expressed specifically in the seminal vesicles, were isolated. pSv-1 and pSv-2 hybridized to mRNAs of 1,600 and 3,500 nucleotides in length, respectively. These cDNA sequences, newly isolated in the present study, may provide useful probes for the study of molecular mechanism of androgen-responsive gene expression.

Androgens↗
PubMed2005

Synthetic vision systems: the effects of guidance symbology, display size, and field of view.

Two experiments conducted in a high-fidelity flight simulator examined the effects of guidance symbology, display size, and geometric field of view (GFOV) within a synthetic vision system (SVS). In Experiment 1, 18 pilots flew highlighted and low-lighted tunnel-in-the-sky displays, as well as a less cluttered follow-me aircraft (FMA), through a series of curved approaches over rugged terrain. The results revealed that both tunnels supported better flight path tracking and lower workload levels than did the FMA because of the availability of more preview information. Increasing tunnel intensity had no benefit on tracking and, in fact, degraded traffic awareness because of clutter and attentional tunneling. In Experiment 2, 24 pilots flew a lowlighted tunnel configured according to different display sizes (small or large) and GFOVs (30 degrees or 60 degrees). Measures of flight path tracking and terrain awareness generally favored the 60 degrees GFOV; however, there were no effects of display size. Actual or potential applications of this research include understanding the impact of SVS properties on flight path tracking, traffic and terrain awareness, workload, and the allocation of attention.

Adult↗
PubMed2005

Phosphorylation of synapsin I by cAMP-dependent protein kinase controls synaptic vesicle dynamics in developing neurons.

In developing neurons, synaptic vesicles (SVs) undergo cycles of exo-endocytosis along isolated axons. However, it is currently unknown whether SV exocytosis is regulated before synaptogenesis. Here, we show that cAMP-dependent pathways affect SV distribution and recycling in the axonal growth cone and that these effects are mediated by the SV-associated phosphoprotein synapsin I. The presence of synapsin I on SVs is necessary for the correct localization of the vesicles in the central portion of the growth cone. Phosphorylation of synapsin I by cAMP-dependent protein kinase (protein kinase A) causes the dissociation of the protein from the SV membrane, allowing diffusion of the vesicles to the periphery of the growth cone and enhancing their rate of recycling. These results provide new clues as to the bases of the well known activity of synapsin I in synapse maturation and indicate that molecular mechanisms similar to those operating at mature nerve terminals are active in developing neurons to regulate the SV life cycle before synaptogenesis.

Animals↗
PubMed2006

Protein kinase A-mediated synapsin I phosphorylation is a central modulator of Ca2+-dependent synaptic activity.

Protein kinase A (PKA) modulates several steps of synaptic transmission. However, the identification of the mediators of these effects is as yet incomplete. Synapsins are synaptic vesicle (SV)-associated phosphoproteins that represent the major presynaptic targets of PKA. We show that, in hippocampal neurons, cAMP-dependent pathways affect SV exocytosis and that this effect is primarily brought about through synapsin I phosphorylation. Phosphorylation by PKA, by promoting dissociation of synapsin I from SVs, enhances the rate of SV exocytosis on stimulation. This effect becomes relevant when neurons are challenged with sustained stimulation, because it appears to counteract synaptic depression and accelerate recovery from depression by fostering the supply of SVs from the reserve pool to the readily releasable pool. In contrast, synapsin phosphorylation appears to be dispensable for the effects of cAMP on the frequency and amplitude of spontaneous synaptic currents and on the amplitude of evoked synaptic currents. The modulation of depolarization-evoked SV exocytosis by PKA phosphorylation of synapsin I is primarily caused by calmodulin (CaM)-dependent activation of cAMP pathways rather than by direct activation of CaM kinases. These data define a hierarchical crosstalk between cAMP- and CaM-dependent cascades and point to synapsin as a major effector of PKA in the modulation of activity-dependent SV exocytosis.

Analysis of Variance↗
PubMed2006

Myosin light chain kinase is not a regulator of synaptic vesicle trafficking during repetitive exocytosis in cultured hippocampal neurons.

The mechanism by which synaptic vesicles (SVs) are recruited to the release site is poorly understood. One candidate mechanism for trafficking of SVs is the myosin-actin motor system. Myosin activity is modulated by myosin light chain kinase (MLCK), which in turn is activated by calmodulin. Ca(2+) signaling in presynaptic terminals, therefore, may serve to regulate SV mobility along actin filaments via MLCK. Previous studies in different types of synapses have supported such a hypothesis. Here, we further investigated the role of MLCK in neurotransmitter release at glutamatergic synapses in cultured hippocampal neurons by examining the effects of two MLCK inhibitors, 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine.HCl (ML-7) and wortmannin. Bath application of ML-7 enhanced short-term depression of EPSCs to repetitive stimulation, whereas it reduced presynaptic release probability. However, ML-7 also inhibited action potential amplitude and voltage-gated Ca(2+) channel currents. These effects were not mimicked by wortmannin, suggesting that ML-7 was not specific to MLCK in hippocampal neurons. When SV exocytosis was directly triggered by a Ca(2+) ionophore, calcimycin, to bypass voltage-gated Ca(2+) channels, ML-7 had no effect on neurotransmitter release. Furthermore, when SV exocytosis elicited by electrical field stimulation was monitored by styryl dye, FM1-43 [N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide], the unloading kinetics of the dye was not altered in the presence of wortmannin. These data indicate that MLCK is not a major regulator of presynaptic SV trafficking during repetitive exocytosis at hippocampal synapses.

Action Potentials↗
PubMed2007

Monitoring and imaging the clot during systemic thrombolysis in stroke patients.

Intravenous thrombolysis is the only approved treatment for acute ischemic stroke when administered within the first 3 h of stroke onset. Response to systemic thrombolysis depends on several factors including the location of arterial occlusion, clot characteristics and, ultimately, the embolic source. In the last few years, tremendous progress has been made, resulting in the widespread implementation of noninvasive neurovascular techniques. These imaging modalities are being increasingly performed in the acute stroke setting, without substantial delay, in a large number of centers worldwide. Transcranial Doppler ultrasound provides a unique opportunity to assess several aspects of clot dissolution by means of continuous monitoring of recanalization during and after tissue plasminogen activator administration. This approach allows for the evaluation of patients at the bedside and in real time due to the commencement, timing, speed and degree of artery reopening in addition to allowing the documentation of reocclusion after successful recanalization. Gradient refocused echo susceptibility vessel sign (GRE SVS) magnetic resonance imaging may be particularly useful for the identification of an intravascular thrombus during the acute phase of ischemic stroke; GRE SVS may represent a surrogate of clot composition and differential response to thrombolysis. The increasing availability of advanced neurovascular techniques may, in the near future, improve the design of stroke trials. The capability of these techniques to assess not only tissue viability but also key aspects regarding susceptibility to thrombolysis such as location, amount, composition, and age of the offending clot may improve the safety and efficacy profile of thrombolytic therapy for acute ischemic stroke.

Humans↗
PubMed2001

Transplacental transmission of Toxoplasma gondii in minipigs infected with strains of different virulence.

Infections with the zoonotic protozoan Toxoplasma gondii during pregnancy can result in severe fetal infections. To investigate the use of pigs as animal models for congenital toxoplasmosis, tachyzoites of 5 T. gondii strains, with low to intermediate virulence in mice, were intravenously inoculated into pregnant minipig gilts. Two strains caused abortions of uninfected fetuses following severe disease of the mothers. One strain caused no disease in the gilts but slightly elevated anti-T. gondii antibodies in 2 of 9 fetuses. One strain produced clinical disease with 4 mummified fetuses and 2 full-term, congenitally infected piglets in 1 gilt and no clinical disease but elevated specific fetal antibodies in both piglets of the other gilt. Infection with the fifth strain (SVS-O14), which was considered apathogenic to both pigs and mice based on the clinical course of this and previous experiments, resulted in significant numbers of congenitally infected piglets, as indicated by production of anti-T. gondii antibodies in all 12 fetuses; the parasite was identified in 3 of these fetuses. This pattern of infection indicates that pigs infected with SVS-O14 (or a similar strain) are relevant animal models for studies of transplacental transmission and pathogenesis of congenital toxoplasmosis.

Animals↗
PubMed1993

Cloning and sequence analysis of a cDNA encoding an androgen-dependent mouse seminal vesicle secretory protein.

We report the cloning and sequencing of a new cDNA sequence encoding a protein from the mouse seminal vesicle. An open reading frame of 297 nucleotides encoded a protein of 99 amino acids with a calculated molecular mass of 11.454 kDa. The first 21 amino acids constituted a signal peptide followed by 78 amino acids encoding the secreted protein. The cDNA sequence comprised a 3' untranslated region of 226 bp and the polyadenylation signal AATAAA, 19 bp upstream from the poly(A)+ tail. A high degree of homology was found between this protein and members of the family of seminal vesicle secretory (SVS) proteins, especially rat SVS VI. Northern blot analysis indicated the presence of a 0.7 kb mRNA species in the mRNAs of seminal vesicle tissue. Castration resulted in a marked decrease in the level of the 0.7 kb mRNA encoding the protein, whereas administration of testosterone to castrated males restored the 0.7 kb mRNA.

Amino Acid Sequence↗
PubMed1995

Structure and sequence of a mouse gene encoding an androgen-regulated protein: a new member of the seminal vesicle secretory protein family.

The gene encoding MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids), an androgen-dependent protein specifically expressed in the mouse seminal vesicle, was isolated and sequenced. A mouse genomic library constructed in the lambda EMBL12 vector was screened using a full length cDNA probe. One genomic clone was selected, 7.4 kb of which were shown to contain the whole MSVSP99 gene. The complete sequence of the MSVSP99 gene (1.7 kb), plus 0.8 and 0.3 kb of the 5' and 3' flanking regions respectively, has been determined. The gene is composed of four exons interrupted by three introns. The size range for the four exons is 47-217 bp, while that of introns is 87-615 bp. The transcription start site was identified as an adenine residue located 21 nucleotides upstream from the ATG start codon. Putative TATA and CAAT boxes were identified, along with a number of regions that shared homologies with known regulatory sequences. These included androgen-responsive elements located in the promoter as well as in the gene sequence. Sequence comparisons with other androgen-responsive genes showed strong homologies between the MSVSP99 gene and the seminal vesicle secretory protein (SVS) family genes (rat SVS II, IV, V and VI). Moreover, some regions were found to be conserved between the MSVSP99 gene and the human semenogelin I and II genes.

Amino Acid Sequence↗
PubMed2006

Antagonists of growth hormone-releasing hormone in oncology.

The development of antagonists of growth hormone (GH) - releasing hormone (GH-RH) is reviewed. GH-RH antagonists bind with a high affinity to pituitary receptors for GH-RH and inhibit the release of GH in vitro and in vivo. The main applications of GH-RH antagonists would be for tumor therapy. The antitumor effects of GH-RH antagonists are exerted in part indirectly through the inhibition of the secretion of pituitary GH and the reduction in the levels of hepatic insulin like growth factor (IGF-I). However, principal effects of the GH-RH antagonists are exerted directly on tumors. Antagonists of GH-RH inhibit the proliferation of various cancer cell lines in vitro and suppress in vivo the levels and the expression of mRNA for IGF-I and IGF-II in tumors. In many human cancers, the effects of GH-RH antagonists appear to be due to the blockade of the action of tumoral GH-RH. GH-RH ligand is present in various human cancers indicating that it may be an autocrine/paracrine growth factor. Splice variants (SVs) of GH-RH receptors and pituitary type of GH-RH receptors that might mediate effects of tumoral GH-RH and of GH-RH antagonists were demonstrated in many human cancers. This suggests the presence of a stimulatory loop based on GH-RH and SVs or pituitary type of GH-RH receptors in diverse tumors. It was shown that GH-RH antagonists inhibited the growth of various human cancer lines xenografted into nude mice including mammary, ovarian, endometrial and prostate cancers, small cell lung carcinomas (SCLC) and non-SCLC, renal, pancreatic, gastric and colorectal carcinomas, malignant gliomas, osteosarcomas and Non-Hodgkin's lymphomas. Further development of GH-RH antagonists should lead to potential therapeutic agents for various cancers.

Animals↗
PubMedDate not supplied

Measuring facial swelling using three-dimensional imaging.

Requirements exist clinically for quantitative analysis of facial swelling. A quick and non-invasive measurement of facial volume is desirable. The acquisition of three-dimensional images, corresponding to spatial models of patients' facial surfaces, has been facilitated using a Spatial Vision System (SVS) developed by Thorn EMI Central Research Laboratories (UK). The SVS acquires four video images (two step pairs) and constructs a three-dimensional surface representation using stereo matching of spatial features acquired using 'speckled' illumination. Pre- and post-operative binary volumetric images are then spatially aligned using surface matching algorithms. Accuracy of registration is optimized using Match Volume Excavation. Segmentation of post-operative swelling is achieved by volume differencing in combination with morphological erosion. Volumetric quantification is achieved using a method of three-dimensional region growing, giving the number of connected voxels in the image volume from a user selected seed point. Accuracy of image acquisition has ben quantified by imaging a range of hemispherical phantoms of known volumes. The mean error in repeated measurement of volume was found to be within 3.5%. The perception of temporal changes in facial swelling is highly subjective. Further work is being undertaken to develop and Enhanced Reality visualization tool in which graphical swelling annotations are combined with real-time patient images.

Computer Simulation↗
PubMed2003

Early- and late-state subacute sclerosing panencephalitis: chemical shift imaging and single-voxel MR spectroscopy.

BACKGROUND AND PURPOSE: Subacute sclerosing panencephalitis (SSPE) is a rare, progressive, inflammatory neurodegenerative disease. Our aim was to determine the metabolic abnormalities of brain in early- and late-stage SSPE by using MR spectroscopy and to assess areas of involvement in the early stages when MR imaging findings were normal. METHODS: Children with stage II (n = 3) or III (n = 3) SSPE and 10 healthy, age-matched children underwent MR imaging, multivoxel MR spectroscopy, and short-echo single-voxel MR spectroscopy (SVS). Areas of involvement in the brain were determined with chemical shift imaging. For SVS, 2 x 2 x 2-cm voxels were placed in the frontal subcortical white matter (FSWM) and parieto-occipital white matter (POWM). N-acetylaspartate (NAA)/creatine (Cr), choline (Cho)/Cr, myo-inositol (Ins)/Cr, and NAA/Cho ratios were calculated. RESULTS: Comparisons of NAA/Cr, Cho/Cr, Ins/Cr and NAA/Cho ratios between patients and control subjects showed significant differences in FSWM and POWM (P <.01). In patients with SSPE, NAA/Cr ratios in POWM were significantly less than those in FSWM (P <.01). NAA/Cr ratios in patients with stage II SSPE and those in the control group were not significantly different; this may reflect the absence of neuronal loss. Decreased NAA/Cr, increased Cho/Cr and Ins/Cr ratios, and increased lactate and lipid peaks were found in patients with stage III SSPE. CONCLUSION: MR spectroscopy showed findings suggestive of inflammation in stage II and findings of demyelination, gliosis, cellular necrosis, and anaerobic metabolism in stage III. MR spectroscopy could be a promising technique for early diagnosis and treatment planning in cases of SSPE.

Aspartic Acid↗
PubMed1992

Synaptic vesicle increase correlated to potentiation of transmission at the synapse of the cat superior cervical ganglion in vivo.

Changes in the pattern, number and size of synaptic vesicles during transmitter release were examined in the synapses of the cat superior cervical ganglion (SCG) in vivo in relation to alteration in the amplitude of postganglionic compound action potential (PGP). Stimulation of the preganglionic nerve fibers at 10 Hz caused an increase in the mean amplitude of PGP. It became augmented by approximately 30% compared to control 10-30 sec after starting the stimulation, and then gradually declined to reach a plateau after 4-6 min. This level, about 20% higher in value than control, was sustained until the end of 30 min of stimulation. The nerve terminal was divided into two areas to examine topographically the numerical changes in synaptic vesicles (SVs): zone I on the presynaptic membrane encircled with a diameter equivalent to the active zone length, and zone II occupying the remaining area outside zone I. The synaptic vesicle density in zone I (vesicle number/microns 2) was 96.9 +/- 4.8 (mean +/- S.E.M.) in the unstimulated control ganglia and 128.8 +/- 9.4 (mean +/- S.E.M.) in the ganglia stimulated for 10-30 sec, which was 30% higher in value than control. Then, it decreased slightly reaching a plateau, 20% higher in value than control. The diameter distribution of the SVs showed that their diameters in zone I (56.6 nm mean) were larger than those (51.6 nm mean) in zone II, and that prolonged stimulation induced smaller vesicles in both areas. The results showed that the increase in SV number in zone I correlated well with the elevation of PGP.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed2004

[Possibilities in the differential diagnosis of brain neoplasms using the long and short time sequences of proton magnetic resonance spectroscopy].

UNLABELLED: Currently to perform proton magnetic resonance spectroscopy (1H MRS) with single voxel spectroscopy (SVS) technique long and/or short echo time sequences are used in order to provide complementary information. PURPOSE: The aim of the study was to compare the usefulness of STEAM (time echo, TE, 20 ms) and PRESS (TE 136 ms) sequences in the evaluation of brain neoplasms. MATERIALS AND METHOD: 15 healthy volunteers and 19 patients with brain tumors were examined using 1, 5 T MR scanner. MRI was performed with a standard protocol for brain neoplasms. 1H MRS with the SVS technique was performed after magnetic resonance imaging (MRI) using STEAM 20 and PRESS 136 sequences (voxel size 1 to 8 cc). RESULTS: In the STEAM sequence besides basic metabolites visible in PRESS as N-acetylaspartate, choline and creatine peaks of myoinositol and glutamate were visualized. Data obtained with short TE sequences in conjunction with long TE data were useful and in 5 cases necessary to identify the tumor type. These results were confirmed by pathology. CONCLUSIONS: The use of short TE sequences enables to visualize metabolites important for the diagnosis and differentiation of the tumor type. The combined information drawn out from both long and short TE spectra is also important.

Adult↗
PubMed2004

[Multivascular atherosclerosis, the Sarajevo Vascular Study].

INTRODUCTION: We started with Sarajevo Vascular Study (SVS) in 1994 with basic aim to evaluate arterial occlusive disease (AOD) of lower extremities and investigate possible effect of amelioration of risk factors to atherosclerosis regression. In 1996 we expanded SVS to polyvascular atherosclerotic disease, asymptomatic or symptomatic atherosclerotic disease on various vascular beds i.e. lower extremity arterial occlusive disease (AOD), cerebrovascular disease (CVD), coronary artery disease (CAD). We enrolled a total of 1680 pts. in the study but this number has changed depending on war migrations, comorbidity, or paramedical reasons. Follow-up of 10 yrs. had 645 pts, and follow-up of 8 yrs. had 1035 pts. METHOD: We enrolled a total of 1680 pts, 954 pts with symptomatic disease, and 246 pts without atherosclerotic disease but with at least 3/8 multiple risk factors (MRF), and as controls 400 pts without vascular disease and less than 3 MRF. STUDY DESIGN: one center, prospective, consecutive, with evaluation of epidemiological data--gender, age, comorbidity, antropometrical data, hemodynamic data--systolic and dystolic velocities, pulsatility and resistive indices, and morphologic data. Clinical variables--gender, age, hypertension, smoking, hyperlipoproteinemia, diabetes mellitus, obesity and fibrinogen. Score of MRF was calculated as x/8. RESULTS: Out of total of 645 pts (10 yrs. follow-up) 399 pts (62%) had AOD, out of them single AOD had 295 pts (74%), and polyvascular disease 104 pts (26%). 63 (61%) pts of symptomatic group had combination of AOD + CAD, and 41 pts (39%) had AOD + CVD + CAD. In the pts with 8 yrs. follow-up (n-1035) AOD had 548 pts. (53%), single AOD had 334 pts (57%) and polyvascular had 122 pts (39%). Out of symptomatic pts. 71 (58.2%) had AOD + CAD, and 51 (41.8%) had AOD + CAD + CVD. Asymptomatic disease on the very entry period of the study was significant for both groups, p < 0.01. CONCLUSIONS: (i) we found a significant number of asymptomatic atherosclerotic changes on other vascular beds, (ii) score of MRF has correlated with polyvascular disease and with overall outcome (iii) antropomethric, haemodynamic, and morphological parameters of human blood vessels have been measured and systematically documented in Bosnia and Herzegovina.

Arteriosclerosis↗
PubMed2005

Preminary report: initial experience of endovascular laser therapy for varicose veins due to greater saphenous vein incompetence in Thailand.

BACKGROUND: Surgical venous stripping (SVS) is a standard treatment for varicose veins (VV) due to greater saphenous vein incompetence (GSVI) but there are some disadvantages to and risks. Endovascular laser (EVL) has been introduced to overcome these disadvantages. The present study was designed to determine the effectiveness of EVL treatment for these patients. MATERIAL AND METHOD: The patients with VV due to GSVI diagnosed by duplex scan were recruited in the present study. The EVL-procedure was percutaneously approached guiding by ultrasound under monitor anesthetic care (MAC). Postoperative clinical and imaging assessment was assessed. RESULTS: There were 17 limbs with symptomatic VVs in 11 patients. Two patients were admitted for a reason not related to surgery. The others were day cases. There was no postoperative complication except a large echymosis in one case. At 3-month follow-up, no recanalization or recurrence was detected. CONCLUSION: The authors' early results demonstrated that EVL could obliterate VVs due to GSVI and further showed some benefits over SVS. More studies with a longer period of follow-up are needed to further confirm the efficacy of EVL.

Adult↗