Evaluations of Spectral Analysis of in vitro 2D-COSY and 2D-NOESY on Human Brain Metabolites

PURPOSE: To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. MATERIALS AND METHODS: All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% D2O. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. RESULTS: Symmetrical 2D-COSY and 2D-NOESY spectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons (CH3(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,H alpha)) at 2.50ppm and methylene protons (CH2,(3,HB)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons (CH3) were observed in NAA. Cross peaks between the methyl protons (CH3(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons (CH2(3)) at 2.11 ppm and methylene protons (CH2(4)) at 2.35 ppm, and between the methylene protons(CH2 (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. CONCLUSION: The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.

Clinical Analysis of Herpes Simplex Keratouveitis

A retrospective study was done to analyze the clinical characteristics about herpes simplex keratouveitis. We reviewed medical records of 28 patients who had been diagnosed as having herpes simplex keratouveitis at Hanyang University Hospital from May 1987 to January 1998. Mean follow-up period was 14 months. In 28 patients(male 19, female 9), first attack was in 5 patients, recurrent uveitis in 16 patients, and 7 patients were undetermined due to lack of detailed clinical history, Stromal keratitis(intersititial keratitis, kisciform keratitis, and necrotizing keratitis) was present in 23 eyes, superficial keratitis(dendritic keratitis, geographic eyes. Ten eyes had an increased intraocular pressure of 21mmHg or more, with 5 eyes having 31mmHg or more, which were all in cases of recurrence. Posterior synechiae were present in 9 eyes, iris atrophy in 4 eyes. Herpes simplex keratouveitis is prone to recur and frequently associated with herpes simplex keratitis, especially stromal keratitis. It may be associated with increased intraocular pressure, osterior synechiae, iris atrophy, or hypopyon, Therefore it is necessary to follow up persistently.

Whole Layer Photocoagulation on the Rabbit Retina with Indirect Diode Laser Ophthalmoscopy

In the treatment of retinopathy of prematurity, it is necessary to coagulate retina including inner retinal layer to achieve regression. However, diode laser has long wavelength, thus damage to bruch`s membrane or choroid can occur when one attempts to coagulate whole retinal layers. We evaluated whether or not we could coagulate whole retinal layer without damage to Bruch`s membrane or choroid in rabbit eyes with indirect diode laser ophthalmoscopy. We palaced laser spots on retina of pigmented rabbits with various laser powers(150, 300, 500mW) and durations(0.3, 0.5, 1.0 second). Fundus photographs were taken immediately after photocoagulation. then we took fundus photographs and enucleated the eyes at one day, one week, 3 weeks, and 5 weeks after photocoagulation to perform pathologic examination. With laser setting of 150mW and 0.3 second, the coagulated retina revealed damaged outer layer but inner retinal layer was spared. We could coagulate whole retinal layer without any damage to Bruch`s membrane or choroid with laser power of 300mW and 0.3 second duration. In 300mW, 0.5 second laser burns and 500mW, 0.3 second burns, internal limiting membrane remained intact while Bruch`s membrane and whole retinal layers were damaged We found whole layer rabbit retina could be coagulated without damage to Bruch`s membrane and choriocapillaries with indirect diode laser opthalmoscopy. and we think it would be necessary to adjust laser power and duration in clinical situation to minimize possible deep tissue injury by diode laser.

A Case of Metastatic Uterine Cervical Carcinma of the Orbit

In adults, it is known that tetastatic tumor to the eye and its adnexa is rare compared with other sites. Orbital tumors can be classified as primary, secondary, and metastatic. Secondary orbital tumors are those that extend to the orbit from a contiguous structure. Metastatic tumors are those that spread to the orbit from more distant site in the body. and their prognosis for life is poor because of widespread systemic malignancy. We experienced a case of metastatic uterine cervical carcinoma of the orbit in a 33-year-old female patient who came to us with a history of sudden proptosis and decreased vision. Eighteen months earlier, the patient had been diagnosed with squamous cell carcinoma of the uterine cervix, and she had undergone chemotherapy, irradiation and surgical intervention. Lung and parametrium metastases were also found six months previously. We report a case of metastatic uterine cervical cancer of the orbit with a review of the literature.