Mitochondrial DNA variants as genetic risk factors for Parkinson disease.

BACKGROUND AND PURPOSE: Investigation of the relationship between mitochondrial DNA (mtDNA) variants and Parkinson disease (PD) remains an issue awaiting more supportive evidence. Moreover, an affirming cellular model study is also lacking. METHODS: The index mtDNA variants and their defining mitochondrial haplogroup were determined in 725 PD patients and 744 non-PD controls. Full-length mtDNA sequences were also conducted in 110 cases harboring various haplogroups. Cybrid cellular models, composed by fusion of mitochondria-depleted rho-zero cells and donor mitochondria, were used for a rotenone-induced PD simulation study. RESULTS: Multivariate logistic regression analysis revealed that subjects harboring the mitochondrial haplogroup B5 have resistance against PD (odds ratio 0.50, 95% confidence interval 0.32-0.78; P = 0.002). Furthermore, a composite mtDNA variant group consisting of A10398G and G8584A at the coding region was found to have resistance against PD (odds ratio 0.50, 95% confidence interval 0.33-0.78; P = 0.001). In cellular studies, B4 and B5 cybrids were selected according to their higher resistance to rotenone, in comparison with cybrids harboring other haplogroups. The B5 cybrid, containing G8584A/A10398G variants, showed more resistance to rotenone than the B4 cybrid not harboring these variants. This is supported by findings of low reactive oxygen species generation and a low apoptosis rate in the B5 cybrid, whereas a higher expression of autophagy was observed in the B4 cybrid particularly under medium dosage and longer treatment time with rotenone. CONCLUSIONS: Our studies, offering positive results from clinical investigations and cybrid experiments, provide data supporting the role of variant mtDNA in the risk of PD.

[Miao Zunyi - his life, writings and students].

Miao Zunyi was an influential physician in the mid-Qing Dynasty. He was self-taught as he read a great amount of prescription books of traditional Chinese medicine. He was proficient in medical theories but flexible in treatment. It was recorded in Draft of Qing History that Miao Zunyi, Ye Tianshi and Xue Shengbai were named as "the three schools of Wuzhong". He began to write books in his later years. He wrote prefaces to Pulse Causes, Syndrome and Treatment (Mai Yin Zheng Zhi) and Wu Yi Hui Jiang. His existing works include Treatise on Febrile Disease (Shang Han Ji Zhu), Wen Re Lang Zhao, Song Xin Notes and Song Xin Medical Cases. Miao's Medical Cases and Song Xin Tang Yi An Jing Yan Chao. He had many remarkable students, such like Huang Tang, Guan Ding, Miao Song, and Shen Nianzu.

[Yang Shoushan Medical Cases in the Wumen Medical School].

The unique manuscript, Yang Shoushan Medical Cases, is now held by the library of Nanjing University of Chinese Medicine.It is the medical cases collection of Yang Shoushan, a well-known doctor of Suzhou in the late Qing Dynasty.It was found that the number of medical cases and the details of each case recorded in this book were much more than that in his other existing medical writings. It greatly enriches the historical materials for the study of Yang's clinical characteristics and academic thought.Its compiler was Huang Shounan, a physician and calligrapher in Suzhou in the late Qing Dynasty and the early Republic of China.This book was not recorded as a book compiled by Huang Shounan before now. This book was believed to be completed in 1890.

Clinical characteristics of essential tremor in Taiwan: an exploratory-comparative study.

BACKGROUND: There are few large-scale clinical analyses of essential tremor (ET) in Asia. We studied the detailed clinical profile with emphasizing the age of onset, tremor location, specific tremor patterns, and rate of progression (ROP) to delineate the characteristics of Taiwanese ET patients and found the difference between the Taiwanese and the Caucasians ET patients. METHODS: All ET patients fulfilled the Movement Disorders Society diagnosis criteria were investigated with a standardized assessment protocol, which including clinical evaluation, uniform severity scoring, self-reported questionnaires, accelerometry, surface electromyography, and videotaped tremor examination. RESULTS: Of 219 patients recruited from July 2008 to October 2009, 153 completed the study protocol. Their mean age was 58.9 years and 47% were women, and 33.3% had family history (FH). There was bimodal distribution in age of tremor onset in patients without but not in those with FH. Head tremor (HT) was present in 48 of 153 (31%) patients. Patients with HT showed slower tremor frequency and less ROP than those without HT. Sixty-seven (44%) patients presented with intention tremor (IT). Male gender and voice tremor were predictive factors of IT occurrence. CONCLUSIONS: Comparing with the Caucasians, Taiwanese ET patients have different patterns of onset-age distribution and lack of female predominance in ET with HT. However, patients with IT and without HT also progressed more rapid as found in the Caucasian.

A Multi-Scale Approach to Model K+ Permeation Through the KcsA Channel.

K+ channels allow a very efficient passage of K+ ions through the membrane while excluding Na+ ions, and these properties are essential for life. The 3D structure of the KcsA K+ channel, solved more than 20 years ago, allows to address many relevant aspects of K+ permeation and selectivity mechanisms at the molecular level. Recent crystallographic data and molecular dynamics (MD) studies suggest that no water is normally present inside the selectivity filter (SF), which can instead accommodate four adjacent K+ ions. Using a multi-scale approach, whereby information taken from a low-level simulation approach is used to feed a high-level model, we studied the mechanism of K+ permeation through KcsA channels. More specifically, we used MD to find stable ion configurations under physiological conditions. They were characterized by two adjacent K+ ions occupying the more central positions of the SF (sites S2 and S3), while the other two K+ ions could be found at the external and internal entrances to the SF. Sites S1 and S4 were instead not occupied by K+. A continuum Bikerman-Poisson-Boltzmann model that takes into account the volume of the ions and their dehydration when entering the SF fully confirmed the MD results, showing peaks of K+ occupancy at S2, S3, and the external and internal entrances, with S1 and S4 sites being virtually never occupied by K+. Inspired by the newly found ion configuration in the SF at equilibrium, we developed a simple kinetic permeation model which, fed with kinetic rate constants assessed from molecular meta-dynamics, reproduced the main permeation properties of the KcsA channel found experimentally, including sublinear current-voltage and saturating conductance-concentration relationships. This good agreement with the experimental data also implies that the ion configuration in the SF we identified at equilibrium would also be a key configuration during permeation.

Effects of saxagliptin on ß-cell stimulation and insulin secretion in patients with type 2 diabetes.

AIM: To study the effect of dipeptidyl peptidase-4 (DPP-4) inhibition with saxagliptin on ß-cell function as reflected by the stimulated insulin secretion rate after an enteral glucose load in patients with type 2 diabetes. METHODS: Patients in this randomized, parallel-group, double-blind, placebo-controlled study were drug-naïve, aged 43-69 years, with baseline haemoglobin A1c (HbA1c) 5.9-8.1%. Twenty patients received saxagliptin 5 mg once daily; 16 received placebo. Patients were assessed at baseline and week 12 by intravenous hyperglycaemic clamp (0-180 min, fasting state), and intravenous-oral hyperglycaemic clamp (180-480 min, postprandial state) following oral ingestion of 75 g glucose. Primary and secondary endpoints were percent changes from baseline in insulin secretion during postprandial and fasting states, respectively. Insulin secretion was calculated by C-peptide deconvolution. RESULTS: After 12 weeks, saxagliptin significantly increased insulin secretion percent change from baseline during the postprandial state by an 18.5% adjusted difference versus placebo (p = 0.04), an improvement associated with increased peak plasma concentrations of intact glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide. In the fasting state, saxagliptin significantly increased insulin secretion by a 27.9% adjusted difference versus placebo (p = 0.02). Saxagliptin also improved glucagon area under the curve in the postprandial state (adjusted difference -21.8% vs. placebo, p = 0.03). CONCLUSIONS: DPP-4 inhibition with saxagliptin improves pancreatic ß-cell function in postprandial and fasting states, and decreases postprandial glucagon concentration. Given the magnitude of enhancement of the insulin response in the fasting state, further study into the effect of DPP-4 inhibition on the ß-cell is warranted.

Contribution of glucocerebrosidase mutation in a large cohort of sporadic Parkinson's disease in Taiwan.

BACKGROUND AND PURPOSE: The association between glucocerebrosidase (GBA) mutations and Parkinson's disease (PD) is attracting increased attention worldwide. In patients of Chinese ethnicity, other than the common L444P mutation, a few mutations have been reported. However, the contribution of GBA to PD can be answered only by a thorough investigation of its mutations in a unique large population. METHODS: We enrolled 1747 participants: 967 PD patients and 780 healthy individuals. We screened entire GBA coding regions and exon-intron boundaries in 30 randomly chosen PD patients, followed by testing five variants (L444P, D409H, R120W, L174P, and Q497R) in all participants. The G2385R and R1628P in LRRK2 had been previously studied in almost all participants. RESULTS: In total, 36 patients (3.72%) carried a heterozygous mutant GBA allele (27 L444P, 7 RecNciI, and 2 D409H). Only two controls (0.26%) carried heterozygous GBA mutation (1 L444P and 1 RecNciI). In PD group, the mean age at onset in carriers was younger than in non-carriers. The difference in percentage of mutation frequencies between patients and controls was highly significant for the L444P mutation (P < 0.0001). One L444P carrier was also associated with LRRK2 G2385R variant, but no atypical Parkinsonism was observed. CONCLUSIONS: The present study ascertains that L444P mutation in GBA gene may contribute to an earlier onset of development of PD in Han/Chinese population. Following LRRK2 variants, GBA is the second most frequent mutations indicated for sporadic PD development in the Han/Chinese population. These GBA carriers are associated with an earlier onset of Parkinsonism.

Photoconduction mechanism of oxygen sensitization in InN nanowires.

The photoconduction (PC) mechanism in indium nitride (InN) nanowires (NWs) has been investigated via environment-, temperature-, and power-dependent measurements. The adsorbed oxygen-induced modulation of the surface state is proposed to be the leading factor in the long lifetime or high gain transport and in sensitizing photocurrent generation in the InN NWs. The electron trapping effect by adsorbed oxygen can be verified by the increased activation energy from 33 ± 4 (in vacuum) to 58 ± 2 meV (in oxygen). The observed supralinear power dependence of photocurrent also suggests the presence of acceptor states that influence the carrier recombination behavior and compensate the thermal carriers in the InN NWs. The potential influence of native oxide on the molecule-sensitive PC in this nitride nanomaterial is also inferred.

First Report of Powdery Mildew Caused by Erysiphe diffusa, Oidium neolycopersici, and Podosphaera xanthii on Papaya in Taiwan.

Powdery mildew can be found in most papaya (Carica papaya L.) fields during the winter and spring seasons in Taiwan. It usually causes severe yellowing of the leaf lamina and petiole and serious defoliation. Three types of powdery mildew fungi were isolated from papaya leaves in Chiayi City (23.28°N, 120.28°E) at the beginning of 2008. Conidia of the first one were single, globose, hyaline, and 24 to 36 × 14 to 18 µm (average 30.2 × 15.6 µm) without fibrosin bodies and with straight or occasionally flexuous conidiophores at the base. The second one had short pseudo-chains of two to four conidia which were ellipsoidal to ovoid, hyaline, and 24 to 40 × 12 to 16 µm (average 29.7 × 13.4 µm) without fibrosin bodies. The third type had chains of ellipsoidal conidia that were hyaline, 24 to 28 × 12 to 16 µm (average 26.3 × 14.4 µm) and contained fibrosin bodies. To confirm the identity of the three fungi, the internal transcribed spacer (ITS) region of rDNA was amplified using the primer pairs G1 (5'-TCC GTA GGT GAA CCT GCG GAA GGA T-3')/Ed2 (5'-CGC GTA GAG CCC ACG TCG GA-3'), G1 (5'-TCC GTA GGT GAA CCT GCG GAA GGA T-3')/On2 (5'-TGT GAT CCA TGT GAC TGG AA-3'), and S1 (5'-GGA TCA TTA CTG AGC GCG AGG CCC CG-3')/S2 (5'-CGC CGC CCT GGC GCG AGA TAC A-3'). The alignment of obtained sequences (GenBank Accession Nos. GU358452, 507 bp; GU358451, 580 bp; and GU358450, 455 bp) showed a sequence identity of 100, 99, and 99% with the ITS sequences of Erysiphe diffusa, Oidium neolycopersici, and Podosphaera xanthii (GenBank Accession Nos. FJ378880, EU909694, and GQ927254), respectively. On the basis of morphological characteristics and ITS sequence similarities, these fungi were identified as E. diffusa (Cooke & Peck) U. Braun & S. Takam., O. neolycopersici L. Kiss, and P. xanthii (Castagne) U. Braun & S. Takam., respectively (1,3). Single colonies on papaya leaves infected with powdery mildew were identified in the laboratory and maintained on papaya leaves as inoculum. Pathogenicity was confirmed through inoculations by gently pressing a single colony of each fungus onto leaves of healthy papaya seedlings (cv. Horng-Fe). Five seedlings were inoculated for each fungus and then covered with plastic bags for 2 days. Five noninoculated seedlings served as control. After inoculation, treated plants were maintained separately from the control in different rooms of a greenhouse at 25°C under natural daylight conditions. Seven days after inoculation, typical symptoms of powdery mildew were observed on inoculated plants, but not on noninoculated plants. The same species from diseased lesions following artificial inoculation with each fungus were identified with light microscopy. Papaya was previously described as a host to O. caricae Noack in many tropical and subtropical areas of the world including Taiwan (2). However E. cruciferarum, Golovinomyces cichoracearum, Oidiopsis sicula, O. caricae, O. caricae-papayae, O. caricicola, O. indicum, O. papayae, Ovulariopsis papayae, P. caricae-papayae, P. macularis, P. xanthii, and Streptopodium caricae were reported to infect papaya (4). To our knowledge, this is the first report of papaya powdery mildew caused by E. diffusa and O. neolycopersici in the world and the first report of the three fungi found on papaya in Taiwan. References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (2) H. S. Chien and H. L. Wang. J. Agric. Res. China 33:320, 1984. (3) L. Kiss et al. Mycol. Res. 105:684, 2001. (4) J. R. Liberato et al. Mycol. Res. 108:1185, 2004.

[The Ding medical family in Jiangpu in the Ming and the Qing Dynasties].

The Jiangpu Ding family was a Gentry Family with many scholars in Nanjing, running through the Ming and the Qing Dynasties. Successful in both medicine and the imperial examination, talents in various fields emerged in large numbers over more than ten generations. Their practice of medicine began with Ding Zhongbao from the original generation, and the second generation of Ding Yi was promoted from a doctor to a local medical officer. From the fifth generation, Ding Feng, became a famous doctor.However, only Ding Yi and Ding Feng were professional doctors in the whole family lineage. The Ding's were still a Gentry Family in essence. The feature of the family was that the Ding's kept their medical background and interests although there existed no professional doctors after Ding Feng in the middle of the Ming Dynasty. This is because the Ding family expected their heirs to acquire medical skills. The Ding's had a number of medical books handed down, such as The Collection of Prescriptions, The Collection of Jade Letters of Pox Department and The Eight Things of Practicing Medicine.

First Report of Anthracnose on Cucurbitaceous Crops Caused by Glomerella magna in Taiwan.

During the summer and fall of 2006, leaf anthracnose samples were collected from fields of cucumber (Cucumis sativus L.), calabash gourd (Lagenaria siceraria (Molina) Standley), and luffa (Luffa cylindrica (L.) M. Roem.) in southern Taiwan. On cucumber leaves, spots start as water-soaked areas and expand into brown spots. Leaf lesions on calabash gourd and luffa begin as water soaked and then become light brown-to-reddish spots. Centers of lesions sometimes fall out; giving infected leaves a shot-hole appearance. Small pieces (approximately 2 × 2 mm) of diseased leaf tissue from margins of individual lesions were surface disinfected in 1% sodium hypochlorite solution for 1 min, rinsed in sterile water, plated on water agar, and incubated at 25°C. After 4 days, mycelium was isolated, transferred to potato dextrose agar (PDA), and then incubated at 25°C in a 12-h light/darkness regimen. Fast-growing colonies on PDA were white to orange or pink with abundant acervuli but no perithecium. One-celled conidia were ovoid to oblong and 12 to 20 × 4 to 6 (15.9 × 5.0) µm. The morphological traits were identical to those of Colletotrichum magna (teleomorph Glomerella magna Jenkins & Winstead) and clearly distinct from those of C. orbiculare (Berk. & Mont.) Arx (synonym C. lagenarium (Pass.) Ellis & Halst. (conidia were mostly oblong, measuring 7 to 11 × 2 to 6 [9.3 × 4.2] µm, with slow-growing gray colonies) (2,3). Koch's postulates were performed to verify that the isolates were capable of causing anthracnose on cucurbitaceous crops. Pathogenicity tests were conducted in the greenhouse at 25°C under natural daylight conditions. Isolate C0604 was grown on PDA for 14 days and a spore suspension was made (106 spores/ml). Three 14-day-old seedlings at the two- to three-leaf stage of muskmelon (Cucumis melo L. var. reticulatus Naud., cv. Sapphire), squash (Cucurbita moschata Duch., cv. Achen), calabash gourd (cv. Huapu), and luffa (cv. 623) were sprayed with the spore suspension and then covered with plastic bags. Control treatments were sprayed with sterile water. After 2 days, the bags were removed. Typical anthracnose symptoms developed on all inoculated seedlings 7 days after inoculation. G. magna was reisolated from inoculated leaves following the protocol used for the original isolation. Control seedlings developed no symptoms. To confirm the identity of the fungus, PCR amplification and DNA sequencing of the internal transcribed spacer 1 (ITS1)-5.8S-ITS2 of rRNA gene of the isolate C0604 was performed by using ITS1/ITS4 as the PCR and sequencing primers. Sequence analysis of the 558-bp PCR product (GenBank Accession No. GU358453) showed 100% identity to the rRNA sequence of G. magna (GenBank Accession No. DQ003103) (1). PCR amplification of the ITS region was also carried out using species-specific primer GmF (5'- GTG AAC ATA CCT CAA ACG TTG CC -3')/GmR (5'- GGA GGG TCC GCC ACT GTA TTT CG -3') designed in this study. A DNA fragment of approximately 378 bp was amplified from nine isolates of G. magna, whereas no amplification products were obtained from reference cultures of C. gloeosporioides (Penz.) Penz. & Sacc. and C. orbiculare. To our knowledge, this is the first report of G. magna causing anthracnose on cucurbitaceous crops in Taiwan. References: (1) M. Du et al. Mycologia 97:641, 2005. (2) S. F. Jenkins, Jr. and N. N. Winstead. Phytopathology 54:452, 1964. (3) T. A. Zitter et al., eds. Compendium of Cucurbit Diseases. The American Phytopathological Society, St. Paul, MN, 1996.

First Report of Fruit Rot Disease of Mango Caused by Botryosphaeria dothidea and Neofusicoccum mangiferae in Taiwan.

Mango (Mangifera indica L.) is an economically important fruit crop in the tropical and subtropical areas of the world. In southern Taiwan, mango is grown on 18,000 ha of hilly land mainly located in Tainan, Kaohsiung, and Pingtung. Tons (180,000) of mango with a value of NT$6.6 billion (US$206 million) are produced annually. In 2008, mango fruit rot disease was observed 1 week after harvest on 30 to 72% of stored mangoes collected from seven orchards in southern Taiwan. The initial symptom was a small, brown lesion and rot symptoms advanced progressively. Two predominant fungi were isolated from the margin of lesions on acidified potato dextrose agar (PDA with lactic acid, pH 3.8). Isolates of each fungal type were transferred to 2% water agar containing sterilized pine needles and exposed to near UV light to induce sporulation. For the first fungus, conidia obtained from pycnidia were ovate, one-celled, and hyaline, with an average length and width of 12.93 ± 0.93 × 6.98 ± 0.40 µm and an average length/width ratio of 1.85. To confirm the identity of the fungus, PCR amplification by universal primers, ITS1/ITS4, and DNA sequencing of the internal transcribed spacer (ITS1-5.8S-ITS2 rRNA gene cluster) were conducted. The internal transcribed spacer (ITS) sequence of ribosomal DNA of this fungus was analyzed and submitted to GenBank (Accession No. GQ421486). It showed a sequence identity of 100% with Neofusicoccum mangiferae (Syd. & P. Syd.) Crous, Slippers & A. J. L. Phillips) (GenBank Accession No. AY615185). For the second fungus, conidia obtained from pycnidia were fusiform, one-celled, and hyaline, with an average length and width of 22.87 ± 1.32 × 6.42 ± 0.46 µm and a length/width ratio of 3.53. The ITS sequence of ribosomal DNA of this fungus was analyzed and submitted to GenBank (Accession No. GQ421485). It showed a sequence identity of 100% with Botryosphaeria dothidea (Moug.: Fr.) Ces & De Not.) (GenBank Accession No. AY 786321). To test pathogenicity, four mango fruits were wounded with a sterile needle, inoculated with mycelium agar plugs (0.5 mm in diameter) excised from separate monoconidial cultures, and incubated in a plastic box with a 100% relative humidity for 2 days at room temperature. Brown lesions appeared on all wounded sites of each fungus 2 days postinoculation. In control experiments, sterile agar plugs were placed on the wounded mango fruits. These fruits remained completely free from symptoms throughout the experiment. The pathogen was reisolated from the lesions of inoculated fruits and identified as N. mangiferae and B. dothidea, thus fulfilling Koch's postulates. N. mangiferae and B. dothidea have been reported on mango trees in Australia and South Africa (1). To our knowledge, this is the first report of these fungi causing fruit rot of mango in Taiwan. References: (1) B. Slippers et al. Mycologia 97:99, 2005.

The crosstalk between IGF-1R and ER-α in the proliferation and anti-inflammation of nucleus pulposus cells.

OBJECTIVE: Insulin-like growth factors-1 receptor (IGF-1R) and estrogen receptor (ER) are reported to co-express and engage in crosstalk involving in the synergistic effect of various aspects. It is unknown whether this crosstalk exists in the nucleus pulposus (NP) cells. We aimed to investigate the interaction between IGF-1R and ER-α in regulating NP cell proliferation and inflammation response under IGF-1 stimulation. PATIENTS AND METHODS: We analyzed the IGF-1, IGF-1R, and ER-α in different degenerated degree human NP tissues. NP cells were cultured with IGF-1 protein with or without the inhibitor of IGF-1R or ER-α to investigate their effects on the proliferation and inflammation response. In addition, we also upregulated the IFG-1R and ER-α expression by plasmid transfection to investigate the impact on each other. The content of IGF-1, IFG-1R, and ER-α was analyzed by enzyme-linked immunosorbent assay (ELISA). The proliferative cell rate was determined by flow cytometry. Additionally, intracellular collagen-II, p16, PCNA, IL-1ß, IL-6, TNF-α, and MMP-13 expression were also detected. RESULTS: We found IGF-1, IFG-1R, and ER-α content were decreased in higher degenerated NP tissues. IGF-1 protein treatment upregulated the IFG-1R and ER-α expression and promoted NP cell proliferation, collagen-II, and PCNA expression. However, the suppression of IGF-1R (or ER-α) weakened the IGF-1 induced collagen-II expression, proliferation, and anti-inflammation effects on NP cells, decreased ER-α (or IGF-1R) expression, and partly reversed the protective effect of NP cells caused by IGF-1 Similarly, the upregulation of one of IGF-1R and ER-α may increase the other as well. CONCLUSIONS: There is an interaction between IGF-1R and ER-α acts synergistically to promote the proliferation and suppress inflammation in NP cells.

A longitudinal study of Taiwanese sialidosis type 1: an insight into the concept of cherry-red spot myoclonus syndrome.

BACKGROUND AND PURPOSE: Sialidosis type 1 (ST-1) is a neurodegenerative disorder with limited long-term follow-up report. This study is to document the chronological profile of ST-1. METHODS: We perform serial analysis of 17 Taiwanese patients with ST-1 focusing on evolution of clinical features, electrophysiological findings, genetic studies, and neuroimage examinations. RESULTS: All patients had a mutation at 554A-->G in exon 3 of the NEU1 gene causing Ser182Gly substitution. Fifteen patients were homozygous. Two patients were heterozygous with novel mutations, 956C-->T causing Ala319Val in one and 163C-->T causing Gln55stop codon in the other. The neuraminidase activity was markedly decreased in all 11 available patients. Only three patients (17.6%) manifested the macular cherry-red spot. The majority of patients (82.3%) developed full-blown manifestation of myoclonus, ataxia, and seizures within 5 years. Abnormal somatosensory evoked potentials with giant cortical waves were found in all patients. Prolonged P100 peak latency of the visual evoked potentials (VEPs) were found in 16 patients (94.1%) in the early stage even without visual symptoms. CONCLUSION: ST-1 in Taiwanese population illustrates distinct characteristics of phenotype with infrequent cherry-red spot. We suggest to screen the NEU1 mutations in patients presenting action myoclonus with abnormal VEPs, even without macular cherry-red spots.

First Report of Papaya Scab Caused by Cladosporium cladosporioides in Taiwan.

In March of 2008, a leaf scab disease was observed in a papaya (Carica papaya L.) orchard at Guoshing, 24.03°N, 120.51°E, in Nantou County, Taiwan. Infected papayas developed symptoms of numerous, pale green, water-soaked areas, 0.5 to 1.5 mm. Infected leaves gradually turned white to gray on the upper surface and small, circular swellings were observed on the abaxial surface. Lesions may coalesce to cover more than 50% of the leaf, rendering them to fall prematurely. Lesions on the lower surface of the leaves were covered with olive-gray patches of mycelia and abundant conidia. Pieces (~2 × 2 mm) of diseased leaf tissue from margins of individual lesions were surface disinfected in 1% sodium hypochlorite solution for 1 min, rinsed in sterile water, plated on water agar, and incubated at 25°C. After 4 days, mycelium was isolated and transferred to potato dextrose agar (PDA). Five isolates (Cc-5 to Cc-9) were isolated and identified as Cladosporium cladosporioides (Fresen.) de Vries based on the velvety, olive-brown with almost black reverse colony color and dimensions and color of conidia and conidiophores (1). Conidia formed in long branched chains that readily disarticulate, single celled, elliptical to limoniform, 2 to 9 (4.6) × 2 to 3 (2.2) µm. Conidia were pale-to-olive brown and smooth to verruculose. Ramoconidia were 0 to 1 septate, 6 to 14 (9.4) × 2 to 4 (2.7) µm, smooth or sometimes minutely verruculose. Conidiophores were pale-to-olive brown, macro- and micronemateus, smooth or sometimes verruculose, 68 to 244 (141.7) × 3.2 to 4 (3.9) µm. To confirm the identity of the fungus, the internal transcribed spacer (ITS) 1 and 4 regions and mitochondrial small subunit (mtSSU) rDNA were sequenced (GenBank Accession Nos. EU935608 and FJ362555), which had 99% homology to the ITS and mtSSU rDNA of C. cladosporioides (GenBank Accession Nos. EU497957 and AY291273, respectively). Pathogenicity tests were conducted in the greenhouse at 25°C with natural daylight conditions. Fungal isolate Cc-6 was used; it was grown on PDA for 6 days and a spore suspension was made (106 spores/ml). Three papaya seedlings (cv. Horng-Fe) were sprayed with the spore suspension and covered with plastic bags. Control treatments were sprayed with sterile water. After 2 days, the bags were removed. Symptoms developed on all inoculated seedlings 4 days after inoculation. In all cases, the typical scab symptom, pale green, water-soaked areas on the lower leaf surface, were observed. C. cladosporioides was reisolated from inoculated leaves following the procedure used for the original isolation. Control seedlings developed no symptoms. The five isolates are being maintained at the DBST, NCYU, Taiwan. Previously, papaya scab reported in China was caused by C. cariciolum Corda (2), C. caricinum C. F. Zhang et P. K. Chi (3), and C. cladosporioides (4). To our knowledge, this is the first report of C. cladosporioides causing papaya scab in Taiwan. References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, England, 1971. (2) H.-H. Peng and Z.-Y. Zhang. J. Yunnan Agric. Univ. 12:23, 1997. (3) C.-F. Zhang. Ph.D. thesis. South China Agricultural University, Guangzhou, P.R.C., 1995. (4) Z. Y. Zhang et al. Flora Fungorum Sinicorum 14:1, 2003.

First Report of a Fruit Rot Disease of Avocado Caused by Neofusicoccum mangiferae.

Production of avocado (Persea americana) has increased significantly during the last 10 years in Taiwan and the area of cultivation is approximately 500 ha. The most important postharvest disease of avocado is anthracnose caused by Colletotrichum gloeosporioides (Penz.) in Taiwan (1). In 2008, a new disease was found to be infecting avocado fruit at some orchards in Tainan County of southern Taiwan. Infected avocados developed smooth, brown, circular spots first on the surface of harvested fruits. A fungus was always isolated from the margin of lesions and could also be found from symptomless fruit pedicles and stems. Fungal colonies cultured on acidified potato dextrose agar (PDA with lactic acid; pH 3.8) were initially colorless, turned dark gradually, and ultimately became gray to dark gray. After 4 days under fluorescent light at 25°C, pycnidia formed on PDA. Conidia obtained from fruiting bodies were ovate, one celled, and hyaline, with an average length and width of 12.9 (9.9 to 15.6) × 6.4 (5.2 to 7.2) µm. The internal transcribed spacer (ITS) sequence of ribosomal DNA of this fungus was analyzed and submitted to GenBank (No. EU847427). It showed a sequence identity of 99% with Neofusicoccum mangiferae ((Syd. & P. Syd.) Crous, Slippers & A.J.L. Phillips) (GenBank No. AY615185). Thus, both morphological and molecular results confirmed the isolated fungus as N. mangiferae. Five avocado fruits were used to test the pathogenicity with three different treatment inoculation sites on each fruit. Wounded and unwounded sites on fruit were inoculated with mycelia agar plugs (0.5 mm in diameter) excised from a monoconidial culture and the fruit was kept in a plastic box with high humidity for 2 days at room temperature. Brown lesions appeared on all wounded sites 2 days postinoculation (dpi) and on unwounded sites at 4 dpi. The pathogen was reisolated from the lesions of inoculated fruits and found to be N. mangiferae, thus fulfilling Koch's postulates. In control experiments, sterile agar plugs were placed on the wounded avocado fruits. These fruits remained completely free from symptoms throughout the experiment. Several species of Botryosphaeria have been reported on avocado, including N. parvum (anamorph of B. parva), Fusicoccum aesculi (anamorph of B. dothidea), and Dothiorella aromatica (= F. luteum). To our knowledge, this is the first report of N. mangiferae causing fruit rot of avocado in Taiwan. Previously, N. mangiferae has been reported on mango trees worldwide, especially in Australia and Thailand (2). The presence of N. mangiferae in the subtropical area presents a serious disease problem not only to avocado but also to mango. References: (1) Y. P. Tsai, ed. List of Plant Diseases in Taiwan. 4th ed. Taiwan Phytopathological Society, 2002. (2) B. Slippers et al. Mycologia 97:99, 2005.

LncRNA Bmncr alleviates the progression of osteoporosis by inhibiting RANML-induced osteoclast differentiation.

OBJECTIVE: To elucidate whether long non-coding RNA (lncRNA) Bmncr could inhibit RANML-induced osteoclast differentiation, thus alleviating the progression of osteoporosis. MATERIALS AND METHODS: Expression level of lncRNA Bmncr at different stages of osteoclast differentiation was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). After Bmncr overexpression or knockdown in RAW 264.7 cells, expression levels of osteoclast-related genes were detected. Bone marrow mesenchymal stem cells (BMMs) isolated from rats undergoing ovariectomy (OVX) were induced with RANKL (50 ng/mL) and M-CSF (50 ng/mL) for 120 h. TRAP staining was conducted to count the number of TRAP-positive osteoclasts containing more than three nuclei. Bone resorption area of bone fragments was quantitatively analyzed. Osteoporosis model in mice was established. Mice were subjected to MicroCT analyses for recording BMD and BV/TV. The expression level of lncRNA Bmncr in the marrow and spleen of osteoporosis mice was examined. RESULTS: LncRNA Bmncr was lowly expressed in the marrow and spleen of osteoporosis mice. Besides, Bmncr expression gradually downregulated during RANKL-induced in vitro osteoclast differentiation, reaching the lowest level at 72 h. The overexpression of Bmncr reduced the amount of osteoclasts, inhibited bone resorption capacity, and downregulated expression levels of Atp6v0d2, Acp5, Ctr, and Mmp9. Conversely, Bmncr knockdown obtained the opposite trends. CONCLUSIONS: LncRNA Bmncr inhibits RANKL-induced osteoclast differentiation, thus alleviating the progression of osteoporosis.

First Report of Simplicillium lanosoniveum Causing Brown Spot on Salvinia auriculata and S. molesta in Taiwan.

Salvinia spp. are small, floating ferns that grow in long chains of two oval leaves and a root-like third leaf. S. natans (L.) All., a native floating fern distributed in paddy fields, ponds, and ditches in Taiwan, has become critically endangered. Another two exotic species, S. auriculata Aublet (eared salvinia) and S. molesta Mitchell (giant salvinia), are sold in increasing frequency at local flower markets and aquarium shops and pose a serious threat when they find their way into the natural environment. Brown spot of S. auriculata was found in a home aquarium in December 2006 in Chiayi, Taiwan. Symptoms of the disease included many, irregular, dark brown spots on both upper and lower leaf surfaces. Lesions on the upper surface of the leaves were covered with white patches of mycelia and abundant conidia. Small pieces (approximately 2 × 2 mm) of diseased leaf tissue from the margin of individual lesions were surface disinfected in 1% sodium hypochlorite solution for 1 min, rinsed in sterile water, plated on water agar, and incubated at 25°C. Six isolates of the fungus were then isolated and transferred to potato dextrose agar (PDA). Isolate Cs0701 was identified morphologically as Simplicillium lanosoniveum (van Beyma) Zare & W. Gams on the basis of morphology of asexual reproduction structures and rDNA sequence analysis (1). In culture, this fungus formed whitish-to-whitish yellow, pulvinate colonies with matted surfaces. The reverse side of cultures was yellow to light brown. Small, ovate to spherical, hyaline conidia, 2.2 to 3.0 × 1.6 to 2.0 µm (average 2.4 × 1.9 µm) were formed. To confirm the identity of the fungus, PCR amplification and DNA sequencing of the internal transcribed spacer (ITS1-5.8S-ITS2 rRNA gene cluster) was conducted on isolates Cs0701 and Cs0702. The sequence of the PCR product was compared with sequences of closely related species listed in the GenBank database. Except for a single nucleotide, the ITS sequence of both isolates (480 bp; GenBank Accession No. EU939525) was identical to the rRNA of Simplicillium lanosoniveum (GenBank Accession No. AJ292396). Koch's postulates were performed to confirm the pathogenicity of the fungus on S. auriculata and S. molesta. After 14 days of growth on PDA, a spore suspension of isolate Cs0701 (106 spores per ml) was sprayed onto approximately 5 and 10 g of healthy S. auriculata and S. molesta plants, respectively, floated in 500-ml beakers filled with 300 ml of tap water. All treatments, including controls misted with sterile water, were replicated three times. The beakers were covered with plastic bags and placed in a growth chamber maintained at 25°C with 12-h fluorescent light cycles. After 2 days, the bags were removed. Symptoms developed on all inoculated plants 4 days after inoculation. In all cases, typical brown spots were observed. Simplicillium lanosoniveum was reisolated from all surface-disinfested infected tissues. Control plants developed no symptoms. Six isolates of the fungus are being maintained at the Department of Microbiology and Immunology, National Chiayi University, Taiwan. To our knowledge, this is the first report of Simplicillium lanosoniveum causing brown spot of S. auriculata and S. molesta in Taiwan. Reference: (1) R. Zare and W. Gams. Nova Hedwigia 73:1, 2001.

First Report of Lasiodiplodia Fruit Rot of Jackfruit in Taiwan.

Jackfruit (Artocarpus heterophyllus Lam.) is a tropical fruit that is native to India. Five diseases, including Rhizopus fruit rot and anthracnose fruit rot, have been recorded in Taiwan (2). In 2003, brown lesions were observed on mature or harvested fruits at the Chiayi Agricultural Experiment Branch. The disease caused fruits to collapse and was easily distinguished from anthracnose and Rhizopus fruit rot. In the field, Rhizopus fruit rot was characterized by black flocci sporangia and mycelia covering the flowers and young fruits. Lasiodiplodia fruit rot often occurred on mature or wounded fruit and diseased fruit were covered with gray or black flat mycelia under humid conditions. In the early stage of Lasiodiplodia fruit rot, tiny yellow-brown lesions appeared on the peel. The lesions could rapidly expand to 10 cm in diameter within 5 days and became dark brown with a light margin. The rot symptoms progressed quickly from the peel surface into the sarcocarps that eventually turned black and soft. A fungus was isolated from the margin of the lesions and cultured on acidified potato dextrose agar (PDA) (pH 3.8). The morphology of the fungus was similar to Lasiodiplodia theobromae (Pat.) Griff. & Maubl. (synonym Botryodiplodia theobromae Pat.), which causes the stem-end rot of mango, papaya, and banana in Taiwan. The fungus grew well and produced pycnidia and conidia on PDA. Young conidia were ovate, hyaline, and thin walled without septa. Mature conidia (20 to 28 × 12 to 15 µm) were dark brown and thick walled with one median septum and longitudinal striations. The internal transcribed spacer (ITS) sequence of ribosomal DNA of this fungus was submitted to GenBank (Accession No. EU 407235) and showed 100% sequence identity with that of Botryosphaeria rhodina (anamorph Lasiodiplodia theobromae; GenBank Accession No. DQ458890). On the basis of morphological and molecular criteria, the fungus was identified as L. theobromae (1). Three healthy jackfruit fruits were wounded and inoculated with 2 × 2 mm mycelial agar plugs of the fungus from a monoconidial culture. A sterile agar plug was placed on the wounded site as a control. The fruits were kept in a box to maintain high humidity for 2 days at room temperature. Brown lesions were observed on all inoculated sites 6 days post infection. The pathogen was reisolated from the lesions of inoculated fruits, fulfilling Koch's postulate. The experiment was repeated twice. To our knowledge, this is the first report of L. theobromae causing fruit rot of jackfruit in Taiwan. References: (1) B. C. Sutton. The Coelomycetes. Commonwealth Mycological Institute, Kew, UK, 1980. (2) Y. P. Tsai, ed. List of Plant Diseases in Taiwan. 4th ed. Taiwan Phytopathological Society, 2002.