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Background Over two decades of observations in the field in South East Asia and Hawai‘i suggest that majority of the commercial dairy herds are of black hair coat. Hence a simple study to determine the accuracy of the observation was conducted with two large dairy herds in Hawaii in the mid-1990s. Methods A retrospective study on longevity of Holstein cattle in the tropics was conducted using DairyComp-305 lactation information coupled with phenotypic evaluation of hair coat color in two large dairy farms. Cows were classified into 3 groups: a) black (B, >90%); b) black/white (BW, 50:50) and c) white (W, >90%). Cows with other hair coat distribution were excluded from the study. In farm A, 211 out of 970 cows were identified having 4 or more lactations. In farm B, 690 out of 1,350 cows were identified with 2 or more lactations for the study. Results The regression analyses and the Wilcoxon-Log-rank test for survival probability showed that Holstein cattle with 90% black hair coat had greater longevity compared to Holstein cattle with 90% white hair coat. Conclusions This study suggests that longevity of Holstein cattle in tropical regions was influenced by hair coat color and characteristics.

New Guinea impatiens, Impatiens hawkeri W. Bull, is widely cultivated as a potted plant and garden plant. In July 2013, hundreds of young plants (cv. Fanfare) showing symptoms of leaf spot with approximately 50% incidence were found in polyethylene tunnels in Yongin City, Korea. Leaf spots were circular to oblong, reaching 6 mm or more in diameter. The spots were initially uniformly brown to reddish brown, turning gray with reddish brown margin. Diseased plants defoliated prematurely and were abandoned without marketing due to signs of discoloration and yellowing on leaves. A cercosporoid fungus was consistently observed in association with disease symptoms. Stromata were brown, small, and composed of a few swollen hyphal cells. Conidiophores were emerging through the cuticle, fasciculate (n = 2 to 20), olivaceous to brown, paler toward the apex, straight to mildly curved, geniculate, 30 to 260 μm long, 3.5 to 5 μm wide, 1- to 6-septate, and with conspicuous conidial scars. Conidia were hyaline and acicular. Smaller conidia were straight and longer conidia were mildly curved. Conidia were subacute to obtuse at the apex, truncate to obconically truncate at the base, 2- to 18-septate, 30 to 320 × 3.5 to 5.5 μm, and with thickened, darkened hila at the base. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora fukushiana (Matsuura) W. Yamam. (1). Voucher specimens were housed in the Korea University herbarium (KUS). An isolate from KUS-F27438 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47640). Fungal DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 (4) and sequenced. The resulting sequence of 497 bp was deposited in GenBank (Accession No. KJ620981). This showed >99% similarity with sequence of C. fukushiana (EF600954) on I. balsamina from Korea. Isolate of KACC47640 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 10 days after inoculation, and were identical to the symptoms observed in the field. C. fukushiana was re-isolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on water-inoculated control plants. Previously, leaf spots of Impatiens spp. associated with C. apii, C. balsaminae, and C. fukushiana have been reported (1,2,3). To our knowledge, this is the first report of C. fukushiana on I. hawkeri in Korea. Our observations in the nurseries of I. hawkeri suggest that low humidity with good ventilation as well as plant hygiene in greenhouses might be main strategies for preventing this disease. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, ARS, USDA, retrieved March 25, 2014. (3) J. M. Soares et al. Plant Dis. 93:1214, 2009. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Seongnam Atopy Preventive Project for Children's Happiness; The Korean Academy of Pediatric Allergy and Respiratory Disease

Bacterial gene islands add to the genetic repertoire of opportunistic pathogens. Here, we perform comparative analyses of three Pseudomonas aeruginosa strains isolated sequentially over a 3-week period from a patient with ventilator-associated pneumonia (VAP) who received clindamycin and piperacillin-tazobactam as part of their treatment regime. While all three strains appeared to be clonal by standard pulsed-field gel electrophoresis, whole-genome sequencing revealed subtle alterations in the chromosomal organization of the last two strains; specifically, an inversion event within a novel 124-kb gene island (PAGI 12) composed of 137 open reading frames [ORFs]. Predicted ORFs in the island included metabolism and virulence genes. Overexpression of a gene island-borne putative β-lactamase gene was observed following piperacillin-tazobactam exposure and only in those strains that had undergone the inversion event, indicating altered gene regulation following genomic remodeling. Examination of a separate cohort of 76 patients with VAP for integration at this tRNA lys recombination site demonstrated that patients exhibiting evidence of integration at this site had significantly higher 28-day mortality. These findings provide evidence that P. aeruginosa can integrate, rapidly remodel, and express exogenous genes, which likely contributes to its fitness in a clinical setting.

†‡ § SUMMARY Interspecific reproductive barriers are poorly understood, but are central to the biological species concept. The pre-zygotic barriers between red- and green-fruited species in the tomato clade of the genus Solanum provide a model to better understand these barriers in plants. Compatibility usually follows the SI x SC rule: pollen from self-compatible (SC) red-fruited species is rejected on pistils of the predominantly self-incompatible (SI) green-fruited species, but the reciprocal crosses are compatible. This suggests that the interspecific reproductive barrier may be linked to the intraspecific SI mechanism. However, pollen from the SC red-fruited species is also rejected by SC accessions of green-fruited species that lack S-RNase, a key protein expressed in pistils of SI Solanum species. Thus, multiple mechanisms may contribute to the barrier between red- and green-fruited species. We tested whether an S-RNase-dependent barrier is sufficient for rejection of pollen from red-fruited species by introducing functional S-RNase, HT-A and HT-B genes from SI species into Solanum lycopersicum (cultivated tomato). We found that expressing S-RNase in combination with either HT-A or HT-B in the pistil is sufficient to cause rejection of pollen from all four red-fruited species. Thus, redundant mechanisms must operate side by side to prevent crosses between red- and greenfruited species in the clade, underlining the complexity of interspecific pollination barriers. Our results also have implications for mating system transitions. We suggest that these transitions must occur in a specific sequence, and that the transition from SI to SC also affects interspecific compatibility.

Epichlorohydrin (ECH) is an antifertility agent that acts both as an epididymal toxicant and an agent capable of directly affecting sperm motility. This study identified the time course of apoptotic cell death in rat epididymides after ECH treatment. Rats were administrated with a single oral dose of ECH (50 mg/kg). ECH-induced apoptotic changes were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and its related mechanism was confirmed by Western blot analysis and colorimetric assay. The TUNEL assay showed that the number of apoptotic cells increased at 8 h, reached a maximum level at 12 h, and then decreased progressively. The Western blot analysis demonstrated no significant changes in proapoptotic Bcl-2-associated X (Bax) and anti-apoptotic Bcl-2 expression during the time course of the study. However, phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) and phospho-c-Jun amino-terminal kinase (p-JNK) expression increased at 8–24 h. Caspase-3 and caspase-8 activities also increased at 8–48 h and 12–48 h, respectively, in the same manner as p-p38 MAPK and p-JNK expression. These results indicate that ECH induced apoptotic changes in rat epididymides and that the apoptotic cell death may be related more to the MAPK pathway than to the mitochondrial pathway.

Pseudomonas aeruginosa is an opportunistic pathogen that can, like other bacterial species, exist in antimicrobial resistant sessile biofilms and as free-swimming, planktonic cells. Specific virulence factors are typically associated with each lifestyle and several two component response regulators have been shown to reciprocally regulate transition between biofilm-associated chronic, and free-swimming acute infections. Quorum sensing (QS) signal molecules belonging to the las and rhl systems are known to regulate virulence gene expression by P. aeruginosa. However the impact of a recently described family of novel quorum sensing signals produced by the Pseudomonas Quinolone Signal (PQS) biosynthetic pathway, on the transition between these modes of infection is less clear. Using clonal isolates from a patient developing ventilator-associated pneumonia, we demonstrated that clinical observations were mirrored by an in vitro temporal shift in isolate phenotype from a non-secreting, to a Type III cytotoxin secreting (TTSS) phenotype and further, that this phenotypic change was PQS-dependent. While intracellular type III cytotoxin levels were unaffected by PQS concentration, cytotoxin secretion was dependent on this signal molecule. Elevated PQS concentrations were associated with inhibition of cytotoxin secretion coincident with expression of virulence factors such as elastase and pyoverdin. In contrast, low concentrations or the inability to biosynthesize PQS resulted in a reversal of this phenotype. These data suggest that expression of specific P. aeruginosa virulence factors appears to be reciprocally regulated and that an additional level of PQS-dependent post-translational control, specifically governing type III cytotoxin secretion, exists in this species.

This study was conducted to investigate the effects of replacing whole crop corn silage (WCCS) with whole crop rice silage (WCRS) in the total mixed ration (TMR) on nutrient intake, milk yield and its composition in Holstein cows. The Chucheong rice variety (Oryza sativa L. Japonica) and corn (Pioneer 32 P75) were harvested at yellow-ripe stage and were ensiled in round bales and in trench silos, respectively. Two TMR containing either WCCS or WCRS were prepared. These diets were randomly assigned to 16 midlactating Holstein cows (8 cows per treatment) and were fed for 120 days. The first 20 days were used for dietary adaptation and for the next 100 days daily feed intake, milk yield and its composition were recorded. The pH, lactic acid, NDF, ADF, CP, Ca and P contents were similar in WCRS and WCCS. The DM, ash and EE contents of WCRS were greater compared with WCCS. Nutrient (DM, NDF, TDN and CP) intakes were similar in cows fed WCCS- and WCRS-based TMR. Daily and 4% fat corrected milk yield were not affected by the treatments. Milk composition (percent milk fat, protein, lactose and total solids) was similar in cows fed either WCCS- or WCRSbased TMR. The concentration of milk urea N was greater in cows fed WCRS-based TMR than those fed WCCS-based TMR. In conclusion, round-baled WCRS can replace WCCS in the diet of mid- to late-lactating Holsteins without any deleterious effects on feed consumption, milk yield and its composition. The present findings raise the possibility that WCRS can be used as an alternative roughage source in the diets of dairy cows in countries with surplus rice production.

Background Increased plasminogen activator inhibitor-1 (PAI-1) concentrations are found in bronchoalveolar lavage (BAL) fluids from patients with ventilator-associated pneumonia or acute respiratory distress syndrome. The authors hypothesized that PAI-1 concentrations were associated with increased mortality in patients with either Pseudomonas aeruginosa-induced ventilator-associated pneumonia or tracheobronchial colonization. Methods In a prospective cohort study, daily aspirates from intubated patients were cultured for P. aeruginosa. Positive patients had blind BAL (bBAL) that was processed for biomarker concentrations. Secretion of type III secretion cytotoxins were also analyzed from the P. aeruginosa strains. Results Thirty-three patients were enrolled. Ten of the 33 patients died. bBAL PAI-1 concentrations were significantly increased in nonsurvivors compared with survivors (31.7 vs. 3.4 ng/ml, P = 0.001 for hospital mortality; 35.9 vs. 4.7 ng/ml, P = 0.02 for 28-day mortality). Even when acute respiratory distress syndrome patients were excluded, there was a significant difference between the survivors and nonsurvivors for bBAL PAI-1 concentrations (P = 0.005). Eighty-three percent of P. aeruginosa strains isolated from patients with high concentrations of bBAL PAI-1 also had strains that secreted cytotoxins. Conclusions PAI-1 concentrations in bBALs correlated with mortality in ventilated patients with positive cultures for P. aeruginosa. Elevated bBAL PAI-1 concentrations also correlated with the secretion of type III exotoxins by P. aeruginosa.

We describe the oldest tracks of web-footed birds from the Early Cretaceous in South Korea. The tracks are characterized by a wide divarication angle and a long reversed hallux. The web is semipalmate and restricted to the proximal portion of the three forward digits. The tracks from the Early Cretaceous in South Korea are smaller than those of the Late Cretaceous, therefore confirming the trend of size increasing in the early evolution of birds as shown by skeletal fossils. The discovery of web-footed tracks with abundant non-web-footed tracks indicates that there was a considerable diversification of shore birds as early as the Early Cretaceous.

Pancreatic islets from DBA/2 mice infected with the D variant of encephalomyocarditis (EMC-D) virus revealed lymphocytic infiltration with moderate to severe destruction of pancreatic beta cells. Our previous studies showed that the major population of infiltrating cells at the early stages of infection is macrophages. The inactivation of macrophages prior to viral infection resulted in the prevention of diabetes, whereas activation of macrophages prior to viral infection resulted in the enhancement of beta-cell destruction. This investigation was initiated to determine whether macrophage-produced soluble mediators play a role in the destruction of pancreatic beta cells in mice infected with a low dose of EMC-D virus. When we examined the expression of the soluble mediators interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in the pancreatic islets, we found that these mediators were clearly expressed at an early stage of insulitis and that this expression was evident until the development of diabetes. We confirmed the expression of these mediators by in situ hybridization with digoxigenin-labelled RNA probes or immunohistochemistry in the pancreatic islets. Mice treated with antibody against IL-1beta or TNF-alpha or with the iNOS inhibitor aminoguanidine exhibited a significant decrease in the incidence of diabetes. Mice treated with a combination of anti-IL-1beta antibody, anti-TNF-alpha antibody, and aminoguanidine exhibited a greater decrease in the incidence of disease than did mice treated with one of the antibodies or aminoguanidine. On the basis of these observations, we conclude that macrophage-produced soluble mediators play an important role in the destruction of pancreatic beta cells, resulting in the development of diabetes in mice infected with a low dose of EMC-D virus.

Bacterial communities in the airways of cystic fibrosis (CF) patients are, as in other ecological niches, influenced by autogenic and allogenic factors. However, our understanding of microbial colonization in younger versus older CF airways and the association with pulmonary function is rudimentary at best. Using a phylogenetic microarray, we examine the airway microbiota in age stratified CF patients ranging from neonates (9 months) to adults (72 years). From a cohort of clinically stable patients, we demonstrate that older CF patients who exhibit poorer pulmonary function possess more uneven, phylogenetically-clustered airway communities, compared to younger patients. Using longitudinal samples collected form a subset of these patients a pattern of initial bacterial community diversification was observed in younger patients compared with a progressive loss of diversity over time in older patients. We describe in detail the distinct bacterial community profiles associated with young and old CF patients with a particular focus on the differences between respective “early” and “late” colonizing organisms. Finally we assess the influence of Cystic Fibrosis Transmembrane Regulator (CFTR) mutation on bacterial abundance and identify genotype-specific communities involving members of the Pseudomonadaceae, Xanthomonadaceae, Moraxellaceae and Enterobacteriaceae amongst others. Data presented here provides insights into the CF airway microbiota, including initial diversification events in younger patients and establishment of specialized communities of pathogens associated with poor pulmonary function in older patient populations.

The type III secretion system of Pseudomonas aeruginosa, responsible for acute infection, is composed of over twenty proteins that facilitate cytotoxin injection directly into host cells. Integral to this process is production and secretion of PcrV. Administration of a recently developed, anti-PcrV immunoglobulin, either as a therapeutic or prophylactic has previously demonstrated efficacy against laboratory strains of P. aeruginosa in a murine model. To determine if this therapy is universally applicable to a variety of P. aeruginosa clinical isolates, genetic heterogeneity of pcrV was analyzed among strains collected from three geographically distinct regions; United States, France and Japan. Sequence analysis of PcrV demonstrated limited variation among the clinical isolates examined. Strains were grouped according to the presence of non-synonymous single nucleotide polymorphisms. Representative isolates from each mutant group were examined for the ability of anti-PcrV to bind the protein secreted by these strains. The protective effect of anti-PcrV IgG against each strain was determined using an epithelial cell line cytotoxicity assay. The majority of strains tested demonstrated reduced cytotoxicity in the presence of anti-PcrV IgG. This study provides insights into the natural sequence variability of PcrV and an initial indication of the amino acid residues that appear to be conserved across strains. It also demonstrates the protective effect of anti-PcrV immunotherapy against a multitude of P. aeruginosa strains from diverse global regions with a variety of mutations in PcrV.

Glycosylated, membrane-associated E1 (58-kDa) and E2 (47- to 49-kDa) rubella virus proteins and unglycosylated nucleoprotein C (33 kDa), from separately expressed vaccinia virus recombinants, were injected into golden Syrian hamsters. Rubella virus E1 and E2 glycoproteins consistently induced an organ-specific autoimmune disease, autoimmune lymphocytic hypophysitis, which was evidenced by the induction of autoantibodies against pituitary cells and by lymphocytic infiltration of the pituitary. Neonatal thymectomy prevented the disease. In contrast, rubella virus nucleoprotein C did not induce either autoantibodies against pituitary cells or lymphocytic infiltration of the pituitary. This finding raises the possibility that virus-specific protein itself can induce an organ-specific autoimmune disease in certain circumstances.

Pseudomonas aeruginosa is one of the major causative agents of mortality and morbidity in hospitalized patients due to a multiplicity of virulence factors associated with both chronic and acute infections. Acute P. aeruginosa infection is primarily mediated by planktonic bacteria expressing the type III secretion system (TTSS), a surface-attached needle-like complex that injects cytotoxins directly into eukaryotic cells, causing cellular damage. Lipopolysaccharide (LPS) is the principal surface-associated virulence factor of P. aeruginosa . This molecule is known to undergo structural modification (primarily alterations in the A- and B-band O antigen) in response to changes in the mode of life (e.g., from biofilm to planktonic). Given that LPS exhibits structural plasticity, we hypothesized that the presence of LPS lacking O antigen would facilitate eukaryotic intoxication and that a correlation between the LPS O-antigen serotype and TTSS-mediated cytotoxicity would exist. Therefore, strain PAO1 (A + B + O-antigen serotype) and isogenic mutants with specific O-antigen defects (A + B − , A − B + , and A − B − ) were examined for TTSS expression and cytotoxicity. A strong association existed in vitro between the absence of the large, structured B-band O antigen and increased cytotoxicity of these strains. In vivo, all three LPS mutant strains demonstrated significantly increased lung injury compared to PAO1. Clinical strains lacking the B-band O antigen also demonstrated increased TTSS secretion. These results suggest the existence of a cooperative association between LPS O-antigen structure and the TTSS in both laboratory and clinical isolates of P. aeruginosa .

Parathyroid hormone (PTH) is a major regulatory factor in skeletal physiology. However, the molecular mechanism underlying the effects of PTH on bones has yet to be elucidated in detail. Recently, some reports have demonstrated the crucial role of bone vasculature with regard to bone density. Angiopoietin-1 (Ang-1), along with VEGF, has been established as a primary angiogenic regulatory agent. In this study, we have attempted to characterize the effects of PTH (1-34) on Ang-1 expression and signaling molecules, employing primary-cultured human osteoblast-like cells. Quiescent osteoblasts were exposed to PTH (1-34), after which Ang-1 expression was determined at the mRNA and protein levels. Reverse transcription-polymerase chain reaction (RT-PCR) analyses indicated that Ang-1 mRNA expression increased as the result of PTH (1-34) treatment. The expression of the Ang-1 protein was also augmented as the result of treatment with PTH (1-34). An adenylyl cyclase activator, forskolin, was shown to induce Ang-1 mRNA expression, whereas the protein kinase A inhibitor, H-89, blocked the PTH (1-34)-mediated expression of Ang-1 mRNA. These findings indicate that PTH (1-34)-mediated Ang-1 expression involves adenylyl cyclase-protein kinase A dependent signaling. Our observations also show that Ang-1 may perform a crucial role in the effects of PTH (1-34) on bones, possibly involving alterations in bone vasculature.

Mammalian forebrain development requires extensive cell migration for cells to reach their appropriate location in the adult brain. Defects in this migration result in human malformations and neurologic deficits. Thus, understanding the mechanisms underlying normal cell migration during development is essential to understanding the pathogenesis of human malformations. Radial glia are known to support radial cell migration, while axons have been proposed as substrate for some non-radially migrating cells. Herein we have directly tested the hypothesis that axons can support non-radial cell migration. One population of cells known to migrate non-radially is the inhibitory interneurons that move from the ganglionic eminence to the cerebral cortex. We first show that early born GABAergic cells colocalize with TAG-1-positive (TAG-1+) axons, while later born cells colocalize with intermediate weight neurofilament-positive, TAG-1-negative (TAG-1-) processes, suggesting temporal differences in substrate specificities. We next developed an in vitro assay that allows us to observe cell migration on axons in culture. Using this assay we find that early born medial ganglionic eminence-derived interneurons migrate preferentially on TAG-1+ axons, while later born cells only migrate on neurofilament-positive/TAG-1- processes. These data provide the first direct evidence that ganglionic eminence cells migrate on axons and that there is an age-dependent substrate preference. Furthermore, the assay developed and characterized herein provides a robust method to further study the molecular substrates and guidance cues of axonophilic cell migration in neural development.

Feed consumption, bodyweight gain and carcass evaluation of Holstein bulls fed either conventional (CON) or organic (ORG) diets were compared. Animals were randomly assigned to either CON (n = 15) or ORG (n = 15) treatments when 6 months old. Inclusion rates of the ingredients were similar in CON and ORG diets formulated for growing (6–12 months of age) and fattening (13–18 months of age) periods. Ingredients of organic origin were used to formulate ORG growing and fattening diets. The bulls on the CON diet were vaccinated, de-wormed and treated conventionally (with antibiotics) against any sign of illness. No dietary or implant growth promotant was used. The bulls on the ORG diet were vaccinated although they were not de-wormed or treated against illness using conventional methods. Instead they were allowed to recover naturally from illness for 3 days. However, if the illness persisted beyond 3 days, they were treated using conventional methods. Consumption of concentrate and rice straw during growing, fattening and overall periods (6–18 months of age) was the same in bulls fed the CON or ORG diets. Bodyweight gain, total dry matter (concentrate and straw) intake and feed to gain ratio during growing and fattening were not affected by the treatments. Daily feed cost and feed cost for unit gain were greater in bulls fed the ORG diet than those fed the CON diet. Weight of cold carcass and different carcass cuts of the bulls were not affected by the treatment. According to the Korean beef grading system, the carcasses from bulls fed either CON or ORG diets were graded similarly but were of lower quality. The Korean beef quality grade is primarily determined by marbling score. The bulls in this study were slaughtered earlier (at 18 months of age) than the conventional slaughtering age of beef cattle (~30 months) in Korea to produce highly marbled beef. It is concluded that the bulls fed ORG diets performed equally, and produced carcasses of similar quality, to those fed CON diets. However, feed costs for a unit bodyweight gain were significantly greater in the ORG diet compared with the CON diet.

Pancreatic islets from SJL/J mice infected with the D variant of encephalomyocarditis virus (EMC-D virus) showed lymphocytic infiltration with moderate to severe destruction of beta cells. Immunohistochemical staining of the islet sections with several monoclonal antibodies, anti-Mac-1, anti-Mac-2, and F4/80 for macrophages, anti-L3T4 for helper/inducer T cells, and anti-Lyt2 for cytotoxic/suppressor T cells revealed that the major population of infiltrating cells at the early stage of viral infection was Mac-2-positive macrophages. In contrast, macrophages detected by anti-Mac-1 and F4/80 monoclonal antibodies were not found at the early stage of viral infection but were found at intermediate and late stages of viral infection. Helper/inducer T cells and cytotoxic/suppressor T cells also infiltrated the islets at intermediate and late stages of viral infection. Short-term treatment of mice with silica prior to viral infection resulted in an enhancement of beta-cell destruction, leading to the development of diabetes. In contrast, long-term treatment of mice with silica resulted in complete prevention of diabetes caused by a low dose of viral infection and a significant decrease in the incidence of diabetes caused by an intermediate or high dose of viral infection. Furthermore, depletion of macrophages by a specific monoclonal antibody (anti-Mac-2) resulted in a much greater decrease in the incidence of diabetes caused by an intermediate dose of viral infection. However, suppression of helper/inducer T cells and cytotoxic/suppressor T cells, by anti-L3T4 and anti-Lyt2 antibodies, respectively, did not alter the incidence of diabetes. On the basis of these data, it is concluded that macrophages, particularly Mac-2-positive macrophages, play a crucial role in the process of pancreatic beta-cell destruction at the early stage of encephalomyocarditis D virus infection in SJL/J mice.

Repeat breeding of a high yielding cow causes enormous economic loss to the dairy producers. In this study, we examined the effect of estradiol benzoate (EB) or GnRH on follicular wave emergence and follicular development, and on pregnancy rates for a CIDR-based timed AI (TAI) protocol in repeat breeder dairy cows. Repeat breeder cows received a CIDR device (InterAg, Hamilton, New Zealand), with an injection of 1 mg EB (SY Esrone�; Samyang, Seoul, Korea) plus 50 mg progesterone (P4; SY Ovaron�; Samyang) (EB + P4 group, n = 36) or with an injection of 250 �g Gonadorelin (GnRH, Fertagyl�; Intervet, Boxmeer, The Netherlands) (GnRH group, n = 35) at the beginning of the experiment (Day 0). On Day 7, CIDR devices were removed and all cows received PGF2� (Lutalyse�; Pharmacia & Upjohn, Puurs, Belgium). Thereafter, cows in the EB + P4 group received an injection of 1 mg EB on Day 8, and TAI 30 h later; those in the GnRH group received an injection of 250 �g GnRH on Day 9, and TAI 19 h later. In a subset of cows for both groups (15 cows from each group), the ovaries of the cows were examined by transrectal ultrasonography (Sonoace 600 with 7.5 MHz linear-array transducer; Medison Co., Seoul, Korea) every 24 h from Days 0 to 8, on Day 10, and on Day 13 for the EB + P4 group, and every 24 h from Days 0 to 9, on Day 11, and on Day 14 for the GnRH group, to observe changes in ovarian structures (follicles and corpora lutea). Pregnancy was diagnosed 35 to 60 days after TAI, using both ultrasonography and rectal palpation. Data were analyzed using Fisher's exact, Student's t-, and chi-square tests. Follicular wave emergence occurred within 7 days in 12/15 EB + P4-treated and 13/15 GnRH-treated cows (P > 0.05). The mean diameters of preovulatory follicles were smaller (P < 0.05) in the EB + P4 group (12.1 � 0.1 mm) than in the GnRH group (13.8 � 0.1 mm). However, the proportion of cows with synchronized ovulation and pregnancy rates to TAI did not differ (P > 0.05) between the EB + P4 (11/15 and 19.4%, respectively) and the GnRH (13/15 and 34.3%, respectively) groups. In conclusion, treatment with either EB or GnRH for a CIDR-based TAI protocol in repeat breeder dairy cows results in synchronous follicular wave emergence, follicular development, and ovulation, and thus similar pregnancy rates to TAI.

Rituximab is a chimeric IgG1 κ monoclonal antibody that targets CD20, a transmembrane phosphoprotein on most B cells. Rituximab depletes B cells by binding to the CD20 molecule and initiating complement-dependent cytolysis or antibody-dependent cell-mediated cytotoxicity.1 Recently, there was a case of muscle-specific kinase (MuSK)-positive myasthenia gravis (MG) successfully treated with rituximab.2 We report a patient with MuSK-positive generalised MG who achieved complete remission with rituximab, after being refractory to steroids, intravenous immunoglobulin, immunosuppressants, thymectomy and less responsive to plasmapheresis. A 32-year-old man was referred for refractory MG. He gave a 2.5-year history of mild bulbar and generalised weakness, mostly in proximal extremities, without sensory symptoms. He denied ptosis and diplopia but reported mild dysphagia and dysarthria. Family history was negative for neuromuscular disorders. He was receiving no medications. He denied any history of smoking and drank socially. The general physical examination was unremarkable. Neurological examination demonstrated diplopia on extreme lateral gaze without fatigable ptosis or Cogan’s lid twitch sign. There was moderate weakness of facial muscles bilaterally and of the tongue, without …

The purpose of this study was to assess the ovarian follicle diameter and hormone status during early postpartum in cows. Eighteen Holstein cows, aged 3.5–5.5 years and of 400–600 kg BW with normal calving, were examined every 3rd day starting from Day 15 until Day 33 postpartum. Blood samples for hormonal (progesterone (P4) and estradiol-17β (E2)) analyses were withdrawn from the coccygeal vein every 3rd day until the end of the experiment. The ovarian follicular number and size were verified and measured using a sector vaginal transducer ultrasound scanner (6.5 MHz probe, SA 600; Medison Co., Seoul, Korea). The least squares means (±SEM) are presented for each day (GLM of SAS). In postpartum cows, E2 levels were 27.09 ± 5.22 pg, 24.50 ± 5.76 pg, 26.92 ± 4.62 pg, 25.53 ± 3.74 pg, 27.47 ± 5.72 pg, 26.48 ± 8.06 pg for Days 15, 18, 21, 24, 27, 30, and 33, respectively, 28.01 ± 7.52 pg; P4 levels were 1.35 ± 1.14 ng, 2.41 ± 1.70 ng, 2.85 ± 2.26 ng, 2.88 ± 1.90 ng, 2.91 ± 1.91 ng, 3.04 ± 2.31 ng, and 4.04 ± 3.70 ng for Days 15, 18, 21, 24, 27, 30, and 33, respectively. The ovarian follicle diameter was 12.36 ± 3.31 mm, 11.91 ± 2.99 mm, 13.09 ± 3.37 mm, 13.55 ± 2.76 mm, 14.55 ± 3.68 mm, 15.64 ± 5.53 mm, 17.45 ± 3.22 mm for Days 15, 18, 21, 24, 30, and 33, respectively. The present results show that mean plasma concentration of P4 and mean follicle diameter gradually increased from Day 15 to Day 33 postpartum. It is concluded that monitoring of P4 and E2 during early postpartum period would be useful to predict the ovarian starts for early rebreeding in dairy cows.