OBJECTIVE: Immunofluorescence microscopy including confocal laser scanning microscopy and electron microscopy were used to study the production of fibronectin, tenascin, and laminin in the cumulus-corona (CC) cells surrounding mature, unfertilized oocytes after ovulation in view of their presumptive importance in the coordination of the processes leading to fertilization and early embryo cleavage, including the final maturation of the ovum, the sperm-egg interaction, and the complex biochemical mechanism between the ovum and the oviduct. METHODS: Mature oocyte-cumulus complex (OCC) was cultured for 24 and 48 hour and fixed in 3.7% formaldehyde. Specimens were incubated with a mixture of primary monoclonal antibodies recognizing different epitopes of fibronectin, tenascin, and laminin, and then with a mixture of secondary antibodies containing FITC, TRITC, and Cy-5 conjugated antibodies. Observation was made by confocal laser scanning microscope equipped with epifluorescece optics. Transmission electron microscopy were used to observe the OCC at 24 and 48 hours after cultrue. RESULTS: The immunocytochemical date demonstrated that CC masses are capable of producing fibronectin and tenascin but their production is heterogeneous in the CC population. Immunoreactivity to fibronectin and tenascin was shown mostly by inner corona cells, and the intensity of immunofluorescence decreased from the central corona cells to the peripheral cumulus cells. Colocalization of fibronectin and tenascin was evident in most CC cells. Moreover, fibronectin and tenascin immunoreactive material was observed in the intracytoplasmic areas, at the plasma membrane level as well as in the extracellular matrix. Whereas, laminin immunofluorescence was found around plasma membrane and extracellular area, but a intracytoplasmic reaction was rarely observed. The distribution of laminin immunofluorescence was similar to that of fibronectin and tenascin, but in some cumulus cells, colocalization between them was not found. Ultrastructurally, cumulus cells projected numerous long, thin microvilli into the intercellular area and some micovilli penetrated into zona pellucida. The inner layer of the cumulus mass was loose arrangement of relatively uniform, small cells with widened intercellular spaces, whereas in the outer layer, cumulus cells are rather larger in size and compact arrangement by narrow, irregular spaces. A small and large linear gap junctions were easily found at cell contacts. The cytoplasm of most cells had abundant organelles typical of steroidogenesis: numerous mitochondrias, a well-developed smooth endoplasmic reticulum, electron dense lipid droplets, and bundles of microtubules and microfilaments. Rudimentary disrupted basal lamina along the cytoplasmic border was rarely seen in a few inner conora cells. CONCLUSION: Even though the functional role of these extracellular matrix proteins remains still unclear, it is reasonable to suggest that they are necessary in various steps of the reproductive process. Cumulus cells appears to be a heterogeneous and dynamic system for suitable microenviroment of fertilization. And functional differences between corona and cumulus cells during the oocyte denudation may be accounted for particular distribution of these adhesive proteins and steroidogenesis-related organelles.
Actin Cytoskeleton ; Adhesives ; Animals ; Antibodies ; Antibodies, Monoclonal ; Basement Membrane ; Cell Membrane ; Cumulus Cells ; Cytoplasm ; Embryonic Structures ; Endoplasmic Reticulum, Smooth ; Epitopes ; Extracellular Matrix ; Extracellular Matrix Proteins ; Extracellular Space ; Female ; Fertilization ; Fibronectins* ; Fluorescein-5-isothiocyanate ; Fluorescent Antibody Technique ; Formaldehyde ; Gap Junctions ; Immunohistochemistry ; Laminin* ; Microscopy, Confocal ; Microscopy, Electron* ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Microtubules ; Microvilli ; Mitochondria ; Oocytes ; Organelles ; Oviducts ; Ovulation ; Ovum ; Sperm-Ovum Interactions ; Tenascin* ; Zona Pellucida

Members of the genus Aspergillus are the most common fungi and all reproduce asexually by forming long chains of conidiospores (or conidia). The impact of various Aspergillus species on humans ranges from beneficial to harmful. For example, several species including Aspergillus oryzae and Aspergillus niger are used in industry for enzyme production and food processing. In contrast, Aspergillus flavus produce the most potent naturally present carcinogen aflatoxins, which contaminate various plant- and animal-based foods. Importantly, the opportunistic human pathogen Aspergillus fumigatus has become the most prevalent airborne fungal pathogen in developed countries, causing invasive aspergillosis in immunocompromised patients with a high mortality rate. A. fumigatus produces a massive number of small hydrophobic conidia as the primary means of dispersal, survival, genome-protection, and infecting hosts. Large-scale genome-wide expression studies can now be conducted due to completion of A. fumigatus genome sequencing. However, genomics becomes more powerful and informative when combined with genetics. We have been investigating the mechanisms underlying the regulation of asexual development (conidiation) and gliotoxin biosynthesis in A. fumigatus, primarily focusing on a characterization of key developmental regulators identified in the model fungus Aspergillus nidulans. In this review, I will summarize our current understanding of how conidiation in two aspergilli is regulated.
Aflatoxins ; Aspergillosis ; Aspergillus ; Aspergillus flavus ; Aspergillus fumigatus ; Aspergillus nidulans ; Aspergillus niger ; Aspergillus oryzae ; Developed Countries ; Food Handling ; Fungi ; Genome ; Genomics ; Gliotoxin ; Humans ; Immunocompromised Host ; Spores, Fungal ; Transcription Factors

Fluoride is one of the most reactive elements in nature, and commonly used in toothpaste since it helps to prevent cavities. Despite this advantage, excessive ingestion of fluoride can cause acute toxicity and gastric disturbance from hydrofluoric acid that was formed in the stomach. We report a case of previously healthy, 41-month-old girl who visited the emergency department with persistent abdominal pain and hematemesis after ingestion of fluoride-containing toothpaste. Though the ingested dose of fluoride was below the toxic dose, serious symptoms developed. We performed esophagogastroduodenoscopy, and confirmed a hemorrhagic gastritis caused by hydrofluoric acid. The girl was uneventfully discharged on day 3 after receiving conservative care. When managing children who ingested fluoride-containing toothpaste, physicians need to consider their symptoms, not the ingested amount. In addition, parents should be cautious when their children use fluoride-containing toothpaste.

Fluoride is one of the most reactive elements in nature, and commonly used in toothpaste since it helps to prevent cavities. Despite this advantage, excessive ingestion of fluoride can cause acute toxicity and gastric disturbance from hydrofluoric acid that was formed in the stomach. We report a case of previously healthy, 41-month-old girl who visited the emergency department with persistent abdominal pain and hematemesis after ingestion of fluoride-containing toothpaste. Though the ingested dose of fluoride was below the toxic dose, serious symptoms developed. We performed esophagogastroduodenoscopy, and confirmed a hemorrhagic gastritis caused by hydrofluoric acid. The girl was uneventfully discharged on day 3 after receiving conservative care. When managing children who ingested fluoride-containing toothpaste, physicians need to consider their symptoms, not the ingested amount. In addition, parents should be cautious when their children use fluoride-containing toothpaste.

Recently xenotransplantation has been thought as a final solution for the controi of donor organ shortage in allograft. In order to be a ciinicai entity, xenotransplantation has many obstacles such as hyperacute rejection and delayed xenogratt rejection as a potent immunologic reaction, zoonosis and ethical problems. We already reported the eariy immunoiogic events occuring soon after xenograft in animal model, in which natural antibody and complement have a crucial roie in rejection response. As a further step for the prolongation of graft survival, we used anticomplement agent (cobra venom factor, CVF) in the same model. Graft survival in discordant (guinea pig-to-rat) xenogratt was extended from 30.6 minutes to 2 days following singie injection of CVF, which showed similar pattern of rejection with the concordant xenogratt in terms of time of rejection response after grafting. In this setting antibody response in the blood did not show any difference between that of pre CVF and post CVF, even though IgM response was more pronounced than IgG. The complement activity in the blood showed marked suppression following CVF injection. Intragraft complement gene (C3 mRNA) expression in CVF injected discordant showed delayed response in a similar pattern like that of concordant xenograft. Interestingly enough intragraft anticomplement gene expression showed the simiiar pattern of response with the complement. From these results we can conclude that anticomplement agent (CVF) extended the graft survival in discordant xenograft upto the level of concordant xenograft by shifting the complement activation response from that of discordant to concordant xenograft.
Rats ; Animals

In this report, a case of a black meniscus with underlying ochronosis is described. Further analysis by laboratory findings showed that the patient had underlying alkaptonuria, which was previously undiagnosed. The patient's symptoms showed improvement after arthroscopic treatment.
Alkaptonuria ; Arthroscopy ; Humans ; Knee ; Ochronosis

Vegetative growth signaling of the opportunistic human pathogenic fungus Aspergillus fumigatus is mediated by GpaA (Galpha). FlbA is a regulator of G protein signaling, which attenuates GpaA-mediated growth signaling in this fungus. The flbA deletion (DeltaflbA) and the constitutively active GpaA (GpaA(Q204L)) mutants exhibit enhanced proliferation, precocious autolysis, and reduced asexual sporulation. In this study, we demonstrate that both mutants also show enhanced tolerance against H2O2 and their radial growth was approximately 1.6 fold higher than that of wild type (WT) in medium with 10 mM H2O2. We performed quantitative PCR (qRT-PCR) for examination of mRNA levels of three catalase encoding genes (catA, cat1, and cat2) in WT and the two mutants. According to the results, while levels of spore-specific catA mRNA were comparable among the three strains, cat1 and cat2 mRNA levels were significantly higher in the two mutants than in WT. In particular, the DeltaflbA mutant showed significantly enhanced and prolonged expression of cat1 and precocious expression of cat2. In accordance with this result, activity of the Cat1 protein in the DeltaflbA mutant was higher than that of gpaA(Q204L) and WT strains. For activity of the Cat2 protein, both mutants began to show enhanced activity at 48 and 72 hr of growth compared to WT. These results lead to the conclusion that GpaA activates expression and activity of cat1 and cat2, whereas FlbA plays an antagonistic role in control of catalases, leading to balanced responses to neutralizing the toxicity of reactive oxygen species.
Aspergillus fumigatus* ; Aspergillus* ; Autolysis ; Catalase ; Fungi ; GTP-Binding Proteins* ; Humans ; Polymerase Chain Reaction ; Reactive Oxygen Species ; RNA, Messenger

PURPOSE: Electrical shock is not always followed by a perfusing rhythm, and multiple shock failure (MSF) is common during CPR. We sought to investigate its risk factors and prognostic implications. METHODS: Adult OHCA patients with emergency department (ED) ACLS attempts were identified from a tertiary hospital OHCA registry extending from 2008 to 2012. Shock failure was defined as any electrical shock attempt not followed by a pulse-generating rhythm. Patients were assigned to one of three groups based on the number of shock failures: 1) MSF: ≥3 electrical shock failures before the first ROSC or CPR termination (if there was no ROSC), 2) early shock success (ESS): pulse-generating rhythm achieved within 3 electrical shock attempts and 3) others: all remaining patients. Baseline characteristics, initial laboratory measurements, and outcomes were compared. RESULTS: A total of 590 patients were included. There was no significant difference in baseline characteristics between the MSF group (n=49) and the early shock success group (n=50) except in its higher proportion of presumed cardiac aetiology. The MSF group showed less severe metabolic acidosis and coagulopathy on ED arrival and better renal function and higher haematocrit and serum albumin levels compared with the ESS group. MSF was associated with less sustained ROSC, but was also associated with more survival discharge and better long-term neurologic outcomes after sustained ROSC. CONCLUSION: MSF may indicate heart-specific problems rather than severe metabolic derangements. Better long-term outcomes can be expected once sustained ROSC is achieved, therefore, this phenomenon warrants more focused research.
Acidosis ; Adult ; Cardiopulmonary Resuscitation ; Emergency Service, Hospital ; Heart Arrest ; Humans ; Resuscitation* ; Risk Factors* ; Serum Albumin ; Shock* ; Tertiary Care Centers ; Ventricular Fibrillation

The traumatic ventricular septal defect (VSD) is a rare but potentially life threatening complication of chest wall injury. The traumatic VSD occurs in up to 4.5% of penetrating cardiac trauma. Most of the patients are usually operated on because of heart failure and/or significant left-to-right shunt. The feasibility of surgical repair under cardiopulmonary bypass may be affected by coexisting pulmonary, cerebral or other vascular injuries. Transcatheter closure of VSD is being considered as an alternative therapeutic modality to surgery in order to avoid the potential risk of cardiopulmonary bypass. We report a patient who underwent a successful transcatheter closure of VSD with an Amplatzer (R) VSD occluder. The patient had a residual VSD with significant left-to-right shunt after surgical repair of post-traumatic VSD using cardiopulmonary bypass.
Cardiopulmonary Bypass ; Heart Failure ; Heart Septal Defects, Ventricular* ; Humans ; Thoracic Wall ; Vascular System Injuries

Posterior fossa epidural hematoma (EDH) is uncommon, but the related clinical deterioration can occur suddenly. Accompanying venous sinus injury and lacerations are associated with 40% to 80% mortality. The authors present one clinical case of a patient with posterior fossa EDH from transverse sinus bleeding. A 57-year-old male was injured after falling while working. He was taken to the hospital, where computed tomography scans of his brain revealed a right posterior temporal and cerebellar EDH with a right temporo-occipital fracture. He underwent a right parieto-occipital craniotomy, incorporating the fracture line. Longitudinal laceration of the right transverse sinus extending to the sigmoid sinus with profuse bleeding was identified. Four gauzes were inserted in the epidural space for tamponade of the injured sinus. Conventional angiography and coil embolization for the injured sinus were immediately performed. Subsequently, the patient was transferred to the operating room, wherein staff members removed the gauzes and remnant hematoma. Based on this experience, the authors recommend that for posterior fossa EDH from transverse sinus bleeding, bleeding control should be performed by gauze packing and endovascular treatment.
Accidental Falls ; Angiography ; Brain ; Colon, Sigmoid ; Cranial Fossa, Posterior ; Craniotomy ; Embolization, Therapeutic ; Epidural Space ; Hematoma ; Hematoma, Epidural, Cranial ; Hemorrhage* ; Humans ; Lacerations ; Male ; Middle Aged ; Mortality ; Operating Rooms