Cysteine-Added Mutants of Turnip Yellow Mosaic Virus

Native turnip yellow mosaic virus (TYMV) is relatively unreactive to maleimide agents, indicating few reactive thiol groups on TYMV. In the present study, we aimed to construct TYMV mutants that have reactive cysteine residues on the surface. To this end, we prepared a library of TYMV mutants where the Thr residue at the C-terminus of coat protein (CP) was replaced by a random sequence of six amino acids that included one cysteine. This library was introduced into Nicotiana benthamiana by agroinfiltration. The CP sequence of the TYMV RNA isolated from inoculated leaves was amplified by reverse transcription-PCR and then used to construct a second library. This process was repeated one more time, and the CP sequences of the TYMV RNA in the inoculated leaves were analyzed. Based on the analysis of over 11,000 CP sequences, the Cys mutants representing most abundant TYMV RNAs were constructed. Analysis of the mutants showed that four Cys mutants were nearly comparable to wildtype with respect to CP and viral RNA levels in N. benthamiana. All these mutants were highly reactive to fluoresceine-5-maleimide. This demonstrates that TYMV can be modified to have additional functional groups on the surface that would be useful for drug delivery.

Production and Storage of Virus Simulants

We have examined isolation and identification protocols for three virus simulant candidates to biological warfare agents. MS2 phage, a simulant for yellow fever virus and Hantaan virus, was propagated using as a host an E. coli strain with F pilus. MS2 phage genome was examined by reverse transcription and polymerase chain reaction (RT-PCR). Coat protein of the phage preparation was examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis. Cydia pomonella granulosis virus (CpGV) is a virus simulant candidate to smallpox virus. CpGV was isolated from a commercialized CpGV pellet. In this study, we developed new isolation and identification protocols for CpGV. One disadvantage of using CpGV is that it is not easy to determine viability of the virus. Here, we have included T4 phage as an alternative. We established a high titer production protocol and developed an easy genome identification protocol that does not require purified phage DNA. Stability of these virus preparations was also examined under various storage conditions. When the virus preparations were not subjected to freeze drying, MS2 phage was most stable when it was stored in liquid nitrogen but unstable at 4℃. In contrast, T4 phage was most stable when it was stored at 4℃. CpGV was stable at −20℃ but not at 4℃. Stability during or after freeze drying was also investigated. The result showed that 70~80% MS2 survived the freeze drying process. In contrast, only about 15% of T4 phage survived during the freeze drying. CpGV was found to be degraded during freeze drying.

Non-electrolyte Solute Permeability of the Rabbit Corneal Epithelium and the Whole Cornea

The permeability in vitro of rabbit corneal epithelium alone and whole cornea to four radioactive substances of various molecular weights-tritiated water (THO: molecular weight 22), mannitol (molecular weight 182), inulin(molecular weight 5,175), and dextran(molecular weight 77,500)-was directly measured by using a newly designed lucite chamber (Fig.1) and a proportional counter system for radioisotope experiments. The permeability of the corneal epithelium and the whole cornea varied with the size of molecular weight (Fig.4) and curvilineally with respect to molecular radius (Fig.5). Certainly the data (Table 2 and 3) support that the greater part of non-electrolyte permeation was through intercellular spaces passively. The epithelial or whole corneal permeability fell markedly as the molecular weight increased from 22 to 5,175. Above the molecular size of inulin, the permeability decreased much as the molecular weight increased. Generally, tritiated water permeability was about 150 times greater than that of mannitol and about 2,000 times than that of inulin and about 10,000 times than that of dextran. The difference between the permeability of the epithelium alone and that of the whole cornea was not noticed. Futhermore, the permeability, either of epithelium alone or the whole cornea, did not vary significantly depending upon the location of the hot side with any of the four kinds of isotope-labelled solutes, whether it was set at the epithelial or endothelial side of the corneal membrane (P>0.1). These results may support that the epithelial layer plays a main role as a physical barrier for nonelectrolyte solutes movement across the rabbit cornea.

Epidemic Hemorrhagic Conjunctivitis in Korea

An unusual form of epidemic conjuctivitis swept through Korea between July and December, 1971. The clinical features of all the cases were uniform and presented marked conjunctival and subconjunctivai hemorrhages in bulbar conjunctiva varied from few minute pin-point petechia to large blotches of frank hemorrhage, and the upper half of the bulbar conjunctiva was invariably the initial site of the hemorahagic lesion, which later spread to the entire bulber conjunctiva, and absent of corneal involvement, such as punctate keratitis, fever and pharyngitis, with rapid and short clinical course of about 10 days. The clinical features strongly suggested that the epidemic hemorrhagic coujunctivitis, which produced subconjunctival hemorrhage instead of punctate keratitis, was presumably caused by a variant type of an adenovirus of epidemic kerato-conjunctivitis.

Effect of Ketalar on Intraocular Pressure and Eye Surgery in Children

Ketalar, a new parenteral anesthetic, was used as a clinical trial for 14 surgical cases ranged from 2 to 15 year old children, and also intraocular pressures of their patient's sound eyes were measured before and after Ketalar injection, I.M. 5-10 mg/kg. Unlike conventional anesthetic agents, Ketalar caused a significant rise in intraocular pressure within 5 minutes and then the pressure improved to normal range about 15 minutes after Ketalar injection. Ketalar could not recommended as a general anethetic agent in case who should examine the intraocular pressure. In eye surgery under 15 year old children, this agent would be a safe and effective general anesthetic, and in an emergent eye injured case, Ketalar is an agent of first choice and can injects to patient regardless of diet intake.

A Case of the Avulsio Bulbi Caused by the Ocular Injury

Authors have recently experienced a rare case of avulsion of the left eye ball due to ocular injury in 7 years old Korean girl. The eye ball was completely exposed out of the lid margin. In this case authors supposed that the dynamic inertia was played a major role in the mechanism of avulsion of this eye ball. A breief review of other literatures has also been described.

Clinical Observation of Corneal Graft: An Interim Report of 25 Keratoplasties

Authors performed 25 corneal grafts in 24 patients since the inauguration of The Central Eye Bank, attached to the Department of Ophthalmology, St. Mary's Hospital, Catholic Medical College, in April 19, 1967. Method and results of these cases are as follows; METHOD: In technique of surgery, the grafts in most of the cases were 7 mm in diameter(Table 3), in two types of penetrating and lamellar corneal grafts(Table 4). The donor material came from patient age group of 51-60 years old in most and was used within 24 hours after death (Table 5). In most cases we placed 12 interrupted sutures except of two cases for continuous suture by 8-0 virgin silks. Preoperative and postoperative cares were routine with systemic dexamethasone, 7.0mg a day was given routinely on the 5 th postoperative day for 30-50 days because of prevention of graft rejection. RESULTS AND SUMMARY: Table 7 summbrizes the results of our observation. Nineteen of the 25 grafts remained clear. And there was improved vision above 20/200 in ten eyes out of nineteen clear grafts. Table 10 lists the reasons why nine clear grafts did not improve vision. Postoperative complications occurred in fourteen grafts (Table 11). Penetrating corneal grafts for adherent leucoma courneae, staphyloma corneae and corneal scars from alkaline burn were failed to maintaining the clear graft. The donor material, the recipient cornea, graft rejection as a complication and its prevantion were discussed. ACKNOWLEDGEMENTS: We wish to express our sincere gratitude to an those who have guided in carrying out the present investigation. In particular, We are jndebted to Dr. Bon Sool Koo, Former Chief and Professor, Department of Ophthalmology, Catholic Medical College. Dr. Sang Wook Rhee. Chief and Associate Professor, Department of Ophthalmology, Catholic Medical College.

Two case of Congenital Anophthalmos

Authors observed clinically for congenital anophthalmos in two cases. One case was a boy, aged 6 years, having congenital anophthalmos of right eye with bifid uvula and another case was newbron baby, aged 4 days, having congenital anophthalmos and blepharophimosis of left eye and was born to undernutciented mother from chronic hepatitis.

Continuous Irrigation in the treatment of the Corneal Alkaline Burn and Serpent Corneal Ulcer

The auther had utilized the continuous irrigation with saline and antibiotics to 1 case of serpent corneal ulcer and 2 cases alkaline burn by Lippas' original technique. Two cases of alkaline burn were healed up without any complications and one case of serpent corneal ulcer showed relatively good result. According to review through a few literature, the method of continuous irrigation is much better one than any other systemic or local administration of the drug to the particular external ocular disease.

A Statistical Observation on the Eye Injuries in the Out-Patients

The authors analysed statistically 223 cases of the eye injuries among 17,547 out-patients who visited to our clinic located at down-town of the city during Jan. 1965 to Sept. 1967. The incidence of the eye injuries is remarkably lower than the other reports from other communities. As far as the objects of eye injuries were concerned, ironpiece, fist and coal-dust were more frequent in 21~40 year-age group, while in under 10 year age group, knife, nail and finger-tip were more frequent. Male was exceedingly prevalent (162) than female (61). 21~30 year age group was also prevalent in regard to the incidence of the injuries.