Effect of supplementation of cryoprotectant solution with hydroxypropyl cellulose for vitrification of bovine oocytes.

BACKGROUND: Successful cryopreservation of bovine oocytes is very important for research and commercial applications. However, the survival and development rate of vitrified-thawed (VT) oocytes are lower than those of non-vitrified-thawed (non-VT) oocytes. OBJECTIVE: To investigate the effect of adding hydroxypropyl cellulose (HPC) to the vitrification solution for bovine oocytes. MATERIALS AND METHODS: For vitrification, bovine metaphase II oocytes were pretreated with a solution containing 10% ethylene glycol supplemented with 0, 10, 50, or 100 ug/mL HPC for 5 min, exposed to a solution containing 30% ethylene glycol supplemented with 0, 10, 50, or 100 ug/mL HPC for 30 s, and then directly plunged into liquid nitrogen. RESULTS: The survival rate of oocytes was significantly higher in the 50 HPC group than in the 0, 10, and 100 HPC groups. The reactive oxygen species level was lower in the non-VT and 50 HPC groups than in the other groups. The mRNA levels of proapoptotic genes (Bax) were lower in the non-VT, 0, and 50 HPC groups than in the other groups. The mRNA levels of antiapoptotic genes (BCl2) were higher in the non-VT than in the other groups. The development rates of embryos (day 8) obtained via parthenogenetic activation (PA) were determined in the non-VT, 0 HPC, and 50 HPC groups. The cleavage rate was significantly higher in the non-VT group. CONCLUSION: Supplementation of vitrification solution with HPC improves the survival of VT bovine oocytes and the development capacity of embryos derived from these oocytes via PA. doi.org/10.54680/fr23110110212.

Investigation of suitable starting doses of narrowband UVB in Asian vitiligo patients: a pilot study.

BACKGROUND: Phototherapy is the preferred treatment modality for active and generalized vitiligo. One of the widely accepted consensus on starting dose of phototherapy is using a uniform dose of 280 mJ/cm2 regardless of patients' Fitzpatrick skin phototype (SPT). However, in many clinical experiences with Asian vitiligo patients, the protocol seems suboptimal. OBJECTIVE: To gather more evidence on establishing a higher starting dosage for Asian vitiligo patients undergoing phototherapy. METHODS: We enrolled generalized vitiligo patients with lesions sized adequate enough for phototest. Minimal erythema dose (MED) of vitiligo lesion and non-lesion was measured along with melanin index (MI). RESULTS: Relatively, a wide range of MED and MI was observed even among patients with similar SPT. The range of MED for lesional skin was 300-700 mJ/cm2 and the MED for non-lesion was 500-800 mJ/cm2 . Correlation was noted between lesional MED and non-lesional MI (Spearman correlation, ρ = 0.664 [P < 0.036]) and mean lesional MED was approximately 65% of mean non-lesional MED. CONCLUSION: Results from phototest and tolerability of patients to doses higher than 280 mJ/cm2 may indicate that higher starting doses might be appropriate for Asian vitiligo patients.

Three cases of pruritic urticarial papules and plaques of pregnancy (PUPPP) treated with intramuscular injection of autologous whole blood.

BACKGROUND: Pruritic urticarial papules and plaques of pregnancy (PUPPP), also known as polymorphic eruption of pregnancy, is a common and benign but exceedingly uncomfortable dermatosis of pregnancy. Investigation of new treatment options has been limited by patient concerns about the negative fetal effects of medication. OBJECTIVE: To assess the efficacy of intramuscular injection of autologous whole blood (AWB) for treatment of PUPPP. METHODS: This is a retrospective descriptive case series of three patients with PUPPP, all of whom were treated with intramuscular injection of AWB. RESULTS: All patients showed good responses to intramuscular injection of AWB, tolerated the treatment, and there were no adverse effects to the patients or their babies. CONCLUSION: AWB may be an alternative treatment option for patients with PUPPP who are worried about the risk of medication use during pregnancy or breastfeeding. Whole blood collected from the patient's own body may be preferable to foreign medications. Future investigation into the exact mechanism with controlled clinical studies using a large number of patients will be necessary to provide supporting evidence for this potential treatment.

Combination treatment by 10 600 nm ablative fractional carbon dioxide laser and narrowband ultraviolet B in refractory nonsegmental vitiligo: a prospective, randomized half-body comparative study.

BACKGROUND: Vitiligo is a common acquired depigmentation disorder caused by the loss of melanocytes. Despite the numerous treatment modalities available for vitiligo, responses to treatment are still unsatisfactory. For this reason, new treatment modalities and approaches are needed. OBJECTIVES: To investigate the effects of fractional carbon dioxide (CO(2) ) laser therapy followed by systemic narrowband ultraviolet B (NB-UVB) phototherapy on nonsegmental vitiligo (NSV) as a prospective and randomized left-right comparative study. METHODS: Ten patients with NSV who presented symmetrical vitiligo lesions with no further improvement despite more than 1 year of conventional treatment were enrolled. Two sessions of half-body fractional CO(2) laser therapy were performed at a 2-month interval. NB-UVB phototherapy was then administered to the entire body 5 days after each fractional laser treatment twice a week, increasing the dose incrementally by 15% at each session. Objective clinical assessments were made by two blinded dermatologists using a quartile grading scale, and the patients' overall satisfaction was evaluated using a 10-point visual analogue scale. RESULTS: Two months after the last treatment, mean improvement scores, assessed by physicians, were significantly higher for those treated with half-body fractional CO(2) laser therapy followed by NB-UVB phototherapy, compared with those treated with NB-UVB alone (P=0·034). In addition, according to subjective assessment, the half-body laser treatment followed by NB-UVB showed significantly higher improvements compared with NB-UVB treatment alone (P=0·023). Noticeable adverse events, such as infection, scarring and Koebner phenomenon, were not found in any patient. CONCLUSIONS: This study suggests that fractional CO(2) laser therapy followed by NB-UVB phototherapy could be used effectively and safely as an alternative modality for the treatment of refractory vitiligo.

Enhancing effect of pretreatment with topical niacin in the treatment of rosacea-associated erythema by 585-nm pulsed dye laser in Koreans: a randomized, prospective, split-face trial.

BACKGROUND: Rosacea is a chronic dermatosis that is usually confined to the face. A pulsed dye laser (PDL) system has been proven to be effective in treating rosacea-associated erythema and telangiectasias. Niacin is a cutaneous vasodilator that can increase the chromophore through increased blood flow. OBJECTIVES: We hypothesized that increased blood flow by pretreatment with topical niacin could enhance the effect of PDL in the treatment of rosacea. METHODS: Eighteen Korean patients with rosacea were recruited. Three sessions of 585-nm PDL using a subpurpuragenic dose with and without pretreatment with niacin cream were performed on randomly assigned half-faces at 3-week intervals. Erythema was assessed objectively by a polarization colour imaging system, and evaluations were also made by three blinded dermatologists. Patient satisfaction was evaluated using a 10-point visual analogue scale. RESULTS: Fifteen patients completed this study. All patients showed an improvement in erythema after three sessions of PDL treatment both with and without niacin pretreatment (P = 0·023 and P = 0·009, respectively). There was no significant difference in the improvement of objective erythema between the two sides. However, based on physician assessment the overall clinical improvement on the niacin side was significantly higher (P = 0·005), and patient satisfaction was also higher on the niacin-pretreated side (P = 0·007). There were no remarkable side-effects, with the exception of transient erythema and oedema. CONCLUSIONS: Pretreatment with topical niacin safely enhanced the effect of 585-nm PDL treatment of rosacea-associated erythema in Koreans. Application of niacin can be helpful in overcoming the relatively lower effect of subpurpuragenic PDL in dark-skinned Asians.

Non-ablative 1550-nm erbium-glass and ablative 10 600-nm carbon dioxide fractional lasers for acne scars: a randomized split-face study with blinded response evaluation.

BACKGROUND: Non-ablative 1550-nm erbium-doped fractional photothermolysis systems (FPS) and 10 600-nm carbon dioxide fractional laser systems (CO(2) FS) have been effectively used to treat scars. OBJECTIVE: We compared the efficacy and safety of single-session treatments of FPS and CO(2) FS for acne scars through a randomized, split-face, evaluator-blinded study. METHODS: Eight patients with acne scars were enrolled in this study. Half of each subject's face was treated with FPS and the other half was treated with CO(2) FS. We used a quartile grading scale for evaluations. RESULTS: At 3 months after the treatment, the mean grade of improvement based on clinical assessment was 2.0 +/- 0.5 for FPS and 2.5 +/- 0.8 for CO(2) FS. On each side treated by FPS and CO(2) FS, the mean duration of post-therapy crusting and scaling was 2.3 and 7.4 days respectively and that of post-therapy erythema was 7.5 and 11.5 days respectively. The mean VAS pain score was 3.9 +/- 2.0 with the FPS and 7.0 +/- 2.0 with the CO(2) FS. CONCLUSION: We demonstrated the efficacy and safety of single-session acne scar treatment using FPS and CO(2) FS in East Asian patients. We believe that our study could be used as an essential reference when choosing laser modalities for scar treatment.

Neuroprotection by extract of Petasites japonicus leaves, a traditional vegetable, against oxidative stress in brain of mice challenged with kainic acid.

BACKGROUND: Reactive oxygen radicals have been implicated in the pathophysiology of many neurologic disorders and brain dysfunctions. Kainic acid has been used as a model agent for the study of neurotoxicity of various excitatory amino acids, since it induces neuronal damage through excessive production of reactive oxygen species. Petasites japonicus MAX (butterbur), cultivated as culinary vegetables in Eastern Asia, contains various kinds of phenolic compounds as well as sesquiterpenes, such as petasin. In European countries, the extracts from roots of Petasites species have been used in the therapy of headache or asthma. AIM OF THE STUDY: The objective of our study is to examine the neuroprotective action of the Petasites japonicus MAX (butterbur) extract against oxidative damage in the brain of mice treated with kainic acid. METHODS: Male ICR mice, 6-8 weeks of age, were administered orally the butanol fraction from methanol extract of Petasites japonicus (BMP) or its subfraction (BMP-I or BMP-II) for 5 consecutive days. Thirty min after the final administration, the animals were challenged s. c. with kainic acid (45 mg/kg), and neurobehavioral activities were monitored. In addition, biomarkers of oxidative stress and neuronal loss in the hippocampus for the biochemical, neurobehavioral,morphological evaluations were analyzed 2 days after the kainic acid challenge. RESULTS: During 5-day treatment with BMP or BMP-1, the body weight gain was not significantly different from that of vehicle- treated control animals. Administration of kainic acid alone induced severe epileptiform seizures, causing a lethality of approximately 50%, and injuries of pyramidal cells in the hippocampus of mice which survived the challenge. Kainic acid exposure also resulted in a remarkable decrease in total glutathione level and glutathione peroxidase activity, and an increase in the thiobarbituric acid-reactive substance (TBARS) value in brain tissues. In comparison, coadministration with BMP (400 mg/kg) reduced the 54% lethality of mice, administered with kainic acid alone, to 25 % (P <0.05). Moreover, BMP at the same dose restored the levels of reduced glutathione and TBARS to control values (P <0.05). In further studies, BMP-I (200 mg/kg) ameliorated significantly (P <0.05) the kainic acid-induced behavioral signs, such as seizure activity, and all mice administered with BMP-I (200 mg/kg) survived the kainic acid toxicity. Consistent with the above, the administration with BMP-1 remarkably attenuated the neurobehavioral signs and neuronal loss in hippocampal CA1 and CA3 regions. CONCLUSION: On the basis of these results, the butanol fraction, especially BMP-I, of Petasites japonicus MAX extract is possibly suggested to be a functional agent to prevent oxidative damage in the brain of mice.

Effects of dissolved calcium and phosphorous on osteoblast responses.

The dissolution behavior of hydroxyapatite (HA) and its effect on the initial cellular response is of both fundamental and clinical importance. In this study, plasma-sprayed HA coatings were characterized by X-ray diffraction and Fourier transform infrared spectroscopy (FTIR). Calcium (Ca) and inorganic phosphorous (Pi) ions released from plasma-sprayed HA coatings within 3 weeks were measured by flame atomic absorption and colorimetrically molybdenum blue complex, respectively. To investigate the effect of dissolution of HA coatings on osteoblast response, additional Ca and Pi were added into the cell culture media to simulate the dissolution concentrations. Human embryonic palatal mesenchyme cells, an osteoblast precursor cell line, were used to evaluate the biological responses to enhanced Ca and Pi media over 2 weeks. Osteoblast differentiation and mineralization were measured by alkaline phosphatase-specific assay and 1,25 (OH)2 vitamin D3 stimulated osteocalcin production. The coatings exhibited an HA-type structure. FTIR indicated the possible presence of carbonates on the coatings. A dissolution study indicated a continual increase in Ca and Pi over time. In the cell culture study, enhanced osteoblast differentiation occurred in the presence of additional Ca concentration in the cell culture media. However, additional Pi concentration in the cell culture media was suggested to slow down osteoblast differentiation and mineralization.

Hepatoprotective and free radical scavenging effects of Nelumbo nucifera.

Ethanol extracts from Nelumbo nucifera (ENN) seeds were studied for possible antioxidative and hepatoprotective effects. Antioxidative effects were measured spectrophotometrically by reduction of 2,2'-Diphenyl-1-picrylhydrazyl (DPPH) radicals. Hepatoprotective effects were tested using carbon tetrachloride (CCl4) and aflatoxin B1 (AFB1)-induced hepatocyte toxicity models. ENN showed potent free radical scavenging effects with a median inhibition concentration of 6.49 microg/ml. Treatment of hepatocytes with ENN inhibited both the production of serum enzymes and cytotoxicity by CCl4. The genotoxic and cytotoxic effects of AFB1 were also inhibited by ENN in dose-dependent manners. These hepatoprotective effects of ENN against CCl4 and AFB1 might result from its potent antioxidative properties.

Clinical and anatomical factors associated with thalamic dyskinesias.

To define the clinical and anatomical factors associated with dyskinesias following thalamic infarction, we performed neurological examination and three-dimensional brain magnetic resonance imaging for 23 patients with thalamic infarction. We measured the total volumes and the largest diameters of the lesions on axial and coronal images. Using the atlas of human thalamus, we investigated the damaged thalamic nuclei. We compared the means of the volumes and the largest diameters of the lesions, and the frequencies of damaged thalamic nuclei between patients with and without thalamic dyskinesias. Seven (two pseudochoreoathetosis and five dystonia) of the 23 patients with thalamic infarction developed dyskinesias. No specific neurological deficits at the onset of stroke predicted the development of dyskinesias. The mean volume of the lesions of patients with dyskinesias (739 mm(3)) was significantly larger than that of those without dyskiensias (92.9 mm(3)). The means of the largest axial (11.6 mm) and coronal (10.8 mm) diameters were significantly larger in patients with dyskinesias, compared to those (axial, 7.1 mm; coronal, 6.4 mm) of patients without dyskinesias. Patients with dyskinesias had damage in the centromedian (CM) thalamic nucleus more frequently compared to those without dyskinesias. Patients with a large thalamic infarction involving the CM nucleus are more likely to develop dyskinesias.

Cloning and characterization of a tobacco cDNA encoding calcium/calmodulin-dependent glutamate decarboxylase.

In plants, gamma-aminobutyric acid (GABA), a major transmitter in the central nervous system in animals, is synthesized by glutamate decarboxylase (GAD), the activity of which is tightly modulated by Ca2+/calmodulin. To study the molecular mechanism of GAD regulation and examine the physiological role of GABA in plants, we isolated and characterized a 1771 bp tobacco cDNA clone, pNtGAD2. The 496 amino acid sequence deduced from pNtGAD2 showed 97, 92, and 81% identity to NtGAD1, petunia, and tomato GAD, respectively. The 26 amino acid sequence within the putative calmodulin binding domain at the carboxy terminus showed a typical alpha-helical structure with hydrophobic and charged amino acid clusters. The pNtGAD2-encoded 56 kDa protein interacted strongly with a monoclonal antibody against the petunia GAD and its GAD activity was stimulated markedly by the addition of exogenous calcium and calmodulin. The molecular sequence of pNtGAD2 and biochemical characteristics of the pNtGAD2-encoded protein confirmed that pNtGAD2 is a clone encoding a functional calmodulin-binding and Ca2+/calmodulin-dependent tobacco glutamate decarboxylase. This result indicates that tobacco plants also have Ca2+/calmodulin-dependent GADs.

Evaluation of chemopreventive effect of dietary selenium-rich egg on mouse skin tumor induced by 2'-(4-nitrophenoxy)oxirane and 12-O-tetradecanoylphorbol-13-acetate.

Chemoprotective effect of dietary selenium (as sodium selenite or as Se-rich egg) on mouse skin tumor induced by topical application of 2'-(4-nitrophenoxy)oxirane (NPO) as tumor initiator and 12-O-tetradecanoylphorbol-13-acetate (TPA) as tumor promoter was evaluated in relation to the dietary source and levels of selenium. Selenium supplementation (0.3 p.p.m.) to the basal diet (0.07 p.p.m. Se) as sodium selenite or as Se-rich egg brought about a 40 or 37% reduction respectively, in the incidence of papilloma formation at 12 weeks after NPO treatment. Tumor yield (number of papillomas per mouse) at 14 weeks after NPO treatment in the basal diet group, basal diet supplemented with 0.3 p.p.m. Se as sodium selenite group and basal diet supplemented with 0.3 p.p.m. Se as Se-rich egg group were 7.5 +/- 2.1, 2.7 +/- 2.3 and 4.1 +/- 3.5 respectively. Dietary supplementation of 1.0 p.p.m. See as Se-rich egg to the basal diet reduced the incidence and the multiplicity of papillomas during the early phase of promotion (11 weeks) but its antitumor activity decreased thereafter, indicating that the accumulation of tissue selenium above the saturated level may not be beneficial. Selenium concentrations in blood, liver and skin tissue of mice in basal diet group (0.33 +/- 0.02, 0.54 +/- 0.10 and 0.21 +/- 0.03 p.p.m. respectively) increased significantly ( P < 0.05) by the supplementation of 0.3 p.p.m. Se as selenite (0.58 +/- 0.02, 1.17 +/- 0.10 and 0.31 +/- 0.05 p.p.m. respectively), 0.3 p.p.m. Se as Se-rich egg (0.59 +/- 0.02, 1.25 +/- 0.11 and 0.33 +/- 0.06 p.p.m. respectively) and 1.0 p.p.m. Se as Se-rich egg (1.20 +/- 0.05, 2.32 +/- 0.28 and 0.51 +/- 0.01 p.p.m. respectively). Glutathione peroxidase activity in blood of mice of the basal diet group (2.4 +/- 0.4 EU/mg) increased significantly (P < 0.05) by dietary selenium supplementation (3.8 +/- 0.6 EU/mg in 0.3 p.p.m. Selenite; 3.7 +/- 0.8 EU/mg in 0.3 p.p.m. Se as Se-rich egg; 4.9 +/- 0.9 EU/mg in 1/0 p.p.m. Se as Se-rich egg) and the enzyme activities in liver and skin tissue also increased by 0.3 p.p.m. Se (as selenite or Se-rich egg) supplementation, but no further increases in their activities were obtained by 1.0 p.p.m. Se (as Se-rich egg). It is, therefore, concluded that a moderate level of dietary selenium (0.3 p.p.m.) has an efficient chemopreventive activity at the promotional stage of carcinogenesis and that dietary selenium rich egg as well as dietary selenite exerted antitumor activity.