Skin collagen can be accurately quantified through noninvasive optical method: Validation on a swine study.

BACKGROUND/PURPOSE: Diffuse reflectance spectroscopy (DRS) is a noninvasive optical technology characterized by relatively low system cost and high efficiency. In our previous study, we quantified the relative concentration of collagen for the individual keloid patient. However, no actual value of collagen concentration can prove the reliability of collagen detection by our DRS system. METHODS: Skin-mimicking phantoms were prepared using different collagen and coffee concentrations, and their chromophore concentrations were quantified using the DRS system to analyze the influence of collagen and other chromophores. Moreover, we used the animal study to compare the DRS system with the collagen evaluation of biopsy section by second-harmonic generation (SHG) microscopy at four different skin parts. RESULTS: In the phantom study, the result showed that coffee chromophore did not severely interfere with collagen concentration recovery. In the animal study, a positive correlation (r=.902) between the DRS system and collagen evaluation with SHG microscopy was found. CONCLUSIONS: We have demonstrated that the DRS system can quantify the actual values of collagen concentration and excluded the interference of other chromophores in skin-mimicking phantoms. Furthermore, a high positive correlation was found in the animal study with SHG microscopy. We consider that the DRS is a potential technique and can evaluate skin condition objectively.

Selenite Stimulates the Proliferation of Intestinal Stem Cells With Elevated Antioxidative Activity.

BACKGROUND: Intestinal stem cells (ISCs) are responsible for the regeneration of intestinal epithelium. In a previous study, we demonstrated that sodium selenite is 1 of the key factors that enhances the growth of ISCs in crypt culture medium. The goal of the present article was to investigate the effect of selenite on the proliferative and antioxidative activities of ISCs. MATERIALS AND METHODS: Five-week old BALB/C mice were administered phosphate-buffered saline or sodium selenite (4 mg/kg/d) for 7 days before ISCs were harvested. The proliferative activity of ISC was indexed by the growth of crypt organoids. The messenger RNA expression levels of ISC markers were quantified by using real-time polymerase chain reaction. The activity of antioxidative enzymes was assayed for glutathione peroxidase (GPx), thioredoxin reductase (TrxR), and superoxide dismutase. RESULTS: Treatment with sodium selenite induced a 1.88-fold increase in the growth number of organoids from ISCs, with elevated expression of leucine-rich repeat-containing G-protein-coupled receptor 5, B lymphoma Moloney murine leukemia virus insertion region homolog-1, and Musashi-1, compared with the ISCs from control samples treated with phosphate-buffered saline. The antioxidative activity of GPx and TrxR was significantly enhanced in the selenite-treated groups (1.55- and 1.23-fold increases, respectively). CONCLUSIONS: Selenite positively regulated the proliferation of ISCs and activated the expression of ISC markers. The elevated activity of GPx and TrxR induced by selenite should promote the antioxidative ability of ISCs and benefit the growth of organoids.

Sonoporation-mediated anti-angiogenic gene transfer into muscle effectively regresses distant orthotopic tumors.

Ultrasound (US) is an effective tool for local delivery of genes into target tumors or organs. In combination with microbubbles, US can temporarily change the permeability of cell membranes by cavitation and facilitate entry of plasmid DNA into cells. Here, we demonstrate that repeated US-mediated delivery of anti-angiogenic genes, endostatin or calreticulin, into muscle significantly inhibits the growth of orthotopic tumors in the liver, brain or lung. US-mediated anti-angiogenic gene therapy also seems to function as an adjuvant therapy that significantly enhances the antitumor effects of the chemotherapeutic drug doxorubicin and adenovirus-mediated cytokine gene therapy. Significantly higher levels of tumor apoptosis or tumor-infiltrating lymphocytes were observed after combined therapy consisting of either anti-angiogenic therapy and chemotherapy, or anti-angiogenic therapy and immunotherapy. Taken together, our experiments demonstrate that intramuscular delivery of anti-angiogenic genes by US exposure can effectively treat distant orthotopic tumors, and thus has great therapeutic potential in terms of clinical treatment.

Deep brain stimulation therapy for Parkinson's disease using frameless stereotaxy: comparison with frame-based surgery.

BACKGROUND AND PURPOSE: Deep brain stimulation (DBS) surgery has been performed using frame-based stereotaxy traditionally; however, in recent years, it has also been performed using frameless stereotaxy. The purpose of this study was to compare the experience at our centre in performing DBS surgery using frameless surgery for patients with Parkinson's disease with that of using frame-based surgery. METHODS: Twenty-four patients with advanced Parkinson's disease underwent DBS surgery, 12 with frameless and 12 with frame-based stereotaxy. After identifying the subthalamus by microelectrode recording (MER), the DBS electrodes were implanted and connected to an implanted programmable generator in all patients. Programming was started 1 month after the operation and the outcome of the patients was followed up regularly for at least 12 months. RESULTS: After 1 year of follow-up, the patients who received frameless surgery showed no difference in the degree of improvement in clinical motor function compared with the patients who received frame-based surgery (P = 0.819); the average improvement was 60.9% and 56.9%, respectively, in the stimulation alone/medication-off state, as evaluated by the Unified Parkinson's Disease Rating Scale-III motor subscore. However, the frameless group had significantly shorter total MER time (P = 0.0127) and a smaller number of trajectories (P = 0.0096) than the frame-based group. CONCLUSIONS: Our data indicate that frameless DBS surgery has a similar outcome when compared with frame-based surgery; however, frameless surgery can decrease the operation time, MER time, and MER trajectory number.

Evaluation of TNF-alpha and IL-1beta polymorphisms in Taiwan Chinese patients with pterygium.

PURPOSE: Pterygium, a complex disease, is associated with ultraviolet radiation, immunoinflammatory process, genetic factors, and virus infection. Ultraviolet radiation induces secretion of proinflammatory cytokines by the ocular surface epithelium, inflammatory cells in the tear fluid, or both. Among these cytokines, tumour necrosis factor (TNF)alpha and interleukin (IL)-1beta activate pterygium body fibroblasts, resulting in a phenotype capable of expressing various proteinases associated with extracellular matrix remodelling, angiogenesis, and fibroblast proliferation, which are important for pterygium formation and recurrence. The genetic factor was proposed to play a role in pterygium formation, but there were few studies to clarify this proposition. For investigating genetic factors, the association between pterygium and TNF-alpha and IL-1beta polymorphisms is evaluated in this study. METHODS: A total of 128 pterygium patients and 103 volunteers without pterygium were enrolled in this study. Polymerase chain reaction-based analysis was used to resolve the TNF-alpha-308 promoter, IL-1beta-511 promoter, IL-1beta exon 5, and IL-1 receptor antagonist (IL-1 Ra) polymorphisms. RESULTS: There were no significant differences in the frequency of genotypes and alleles of TNF-alpha-308 promoter, IL-1beta-511 promoter, IL-1beta exon 5, and IL-1 Ra polymorphisms between both groups. CONCLUSIONS: The correlation between pterygium and TNF-alpha-308 promoter, IL-1beta-511 promoter, IL-1beta exon 5, and IL-1 Ra polymorphisms does not exist and those polymorphisms are not useful genetic markers for pterygium susceptibility. Further studies on other polymorphisms or haplotypes of TNF-alpha and IL-1beta are necessary.

Multiple deep-seated cavernomas in the third ventricle, hypothalamus and thalamus.

BACKGROUND: Cavernomas are rarely located in the third ventricle, hypothalamus, or thalamus. In this report, we present our experience managing a patient with three cavernomas, one each in the floor of the third ventricle, hypothalamus, and left thalamus. CASE PRESENTATION: This 62-year-old woman had had an unsteady gait and weakness of both legs for six months. Magnetic resonance imaging (MRI) revealed multiple intracranial tumours in the third ventricle, hypothalamus, and left thalamus. The third ventricle tumour was found to be a cavernoma by intra-operative endoscopic examination and then was excised via a transcortical, transventricular approach. Pathology revealed a cavernoma. The other two tumours were assumed to be cavernomas because of their MRI features. Three days after surgery, the patient developed right hemiparesis and disturbance of consciousness. Computed tomography revealed a left thalamic haemorrhage. After conservative treatment, her conscious level gradually recovered and she could walk with support seven months after surgery. INTERPRETATION: Our experience with this rare case of multiple, deep-seated cavernomas suggests that management of such patients requires specific consideration of the clinical manifestations, location, size, and previous bleeding history.

Dural metastasis in patients with malignant neoplasm and chronic subdural hematoma.

OBJECTIVES: Dural metastasis associated with chronic subdural hematoma (CSH) is rare in patients with malignant neoplasm. In this study, biopsy of the dura and cytological examination of the subdural hematoma was performed for patients with malignant neoplasm and chronic subdural hematoma to investigate the association of dural metastasis and CSH. MATERIALS AND METHODS: Four patients with malignant neoplasm (one breast, one lung, and one colon cancers, and one lymphoma) were diagnosed with CSH. Biopsy of the dura and cytological examination of the subdural fluid were performed for each of these patients. RESULTS: Pathological examination of the dura revealed metastasis for two patients (one lymphoma and one breast cancer), with no specific change except neomembrane formation revealed for the dura of the other two patients. The cytology study was negative for all four patients. All these four patients died within 10 days of the operation; one from recurrent subdural hematoma, and three from infection. CONCLUSION: The results suggest that dural metastasis should be considered in patients with malignant neoplasm and CSH. Further, the prognosis for patients with malignant neoplasm and CSH may be poor because of systemic metastasis and the side-effects of chemotherapy.

Slipknot for scleral fixation of intraocular lenses.

We describe a simple, quick technique to construct a slipknot that can be snared around the haptic of an intraocular lens (IOL) during transscleral fixation of a posterior chamber IOL or repositioning of a dislocated IOL. An in vitro experiment showed that the time required for the slipknot method was significantly less than that for the traditional triple-knot method. An additional advantage is that the slipknot technique can be performed by a single surgeon using standard ophthalmic instrumentation. In addition, the slipknot is as secure as a triple knot.

Risk factors in endophthalmitis leading to evisceration or enucleation.

BACKGROUND AND OBJECTIVE: This study aimed to evaluate the risk factors associated with evisceration or enucleation in endophthalmitis cases. PATIENTS AND METHODS: A retrospective analysis was conducted of all cases of endophthalmitis in a single unit over a 10-year period. The cases were divided into two groups. The group 1 consisted of patients with endophthalmitis who underwent evisceration or enucleation. The group 2 included patients with endophthalmitis who recovered with intravitreal injection or vitrectomy. RESULTS: During the 10-year study period, 86 patients with endophthalmitis were admitted for treatment. Evisceration or enucleation was undergone by 20 patients (23.2%) (Group 1); Group 2 consisted of 66 patients. Older age (P = 0.03), no light perception at presentation (P < 0.0001), endophthalmitis associated with corneal ulcer (P < 0.0001), and endogenous endophthalmitis (P = 0.048) were statistically significant factors associated with the need for evisceration or enucleation.

Impression cytology study of conjunctival epithelial phenotypes on the healing ocular surface after pterygium excision.

PURPOSE: To compare the process of conjunctival epithelial regeneration after three types of pterygium excision procedures. METHODS: Thirty-eight patients (45 eyes) with primary pterygium were randomly assigned to a bare-sclera procedure (group 1, 15 eyes of 12 patients), bare-sclera with intraoperative mitomycin C (MMC 0.02% for 30 seconds; group 2, 15 eyes of 14 patients), or pterygium excision with conjunctival autografting (group 3, 15 eyes of 12 patients). Controls were healthy fellow eyes and seven eyes of age- and sex-matched subjects. Impression cytology was performed preoperatively, at 1 and 2 weeks, and at 1, 3, 6, and 12 months after surgery. The nucleus-to-cytoplasm (N/C) ratio of nongoblet epithelial cells and goblet cell density (GCD) in the pterygial area were calculated and compared over time across treatment groups. RESULTS: Pterygium excision wounds healed in a similar four-stage process in all groups, but at different rates and with different final results. The N/C ratio was highest at about 1 month postoperatively in groups 1 and 2 and at 2 weeks in group 3, before gradually returning to control levels. Preoperatively, the GCD in treated eyes was almost twice that in control eyes (p = 0.001) but fell to zero immediately postoperatively. Goblet cells first appeared (with the most rapidly increased density) in group 3, followed by group 1. At 12 months, the mean GCD in groups 1 and 3 were not significantly different from those in controls, whereas the mean GCD in group 2 was still less than that of control (p = 0.02). CONCLUSIONS: Healing of conjunctiva is delayed by MMC and is promoted by autografting. Even 1 year after surgery, the ocular surface remains abnormal with respect to epithelial phenotypes in eyes treated by any of the three techniques.

Low-dose intraoperative mitomycin C as chemoadjuvant for pterygium surgery.

PURPOSE: To evaluate the efficacy and safety of low-dose intraoperative mitomycin C (MMC) during bare sclera procedure and to compare the rates of pterygium recurrence between recurrent pterygium patients treated with adjuvant MMC and those reconstructed with a conjunctival autograft. METHODS: We studied the recurrence of pterygium, 12-month pterygium-free survival rates, final appearance, and postoperative complications in 96 eyes of 92 patients who received either intraoperative 0.02% MMC for 30 seconds or conjunctival autografting after pterygium excision. Patients were divided into three groups: group 1 included 38 eyes with primary pterygium undergone intraoperative MMC; group 2, 26 eyes with recurrent pterygium treated with intraoperative MMC; and group 3, 32 eyes with recurrent pterygium treated with pterygium excision and conjunctival autografting. RESULTS: Pterygium recurred in 3 (7.9%) of 38 eyes in group 1, 5 (19.2%) of 26 eyes in group 2, and 2 (6.3%) of 32 eyes in group 3. Despite the higher recurrence rate in group 2 compared with that of group 3, the difference between the two was not statistically significant (p = 0.22). The cumulative probabilities of success were 91.6+/-4.6%, 80.8+/-7.7%, and 92.3+/-5.4% at 12 months for groups 1, 2, and 3, respectively. Final appearance of the pterygium excision area was satisfactory in nearly two-thirds of the MMC-treated patients, 71.1% and 65.4% for groups I and 2, compared to 75.1% of patients who had undergone conjunctival autografting. No patients experienced severe complications during a mean postoperative follow-up of 27.3+/-4.1 months, 29.9+/-3.9 months, and 40.9+/-19.1 months for groups 1, 2, and 3, respectively. CONCLUSIONS: A single intraoperative application of 0.02% MMC for 30 seconds after pterygium excision is associated with minimal complication and effectively reduces the recurrence rates after excision of primary or recurrent pterygium. In comparison with conjunctival autografting, low-dose application of MMC after bare sclera procedure is less efficacious in preventing recurrence of pterygium, but simpler and produces a similar proportion of patients with satisfactory final appearance.

Repositioning a subluxated sutured intraocular lens in a vitrectomized eye.

A simple, effective technique for repositioning a subluxated intraocular lens (IOL) in a vitrectomized eye is reported. A 49-year-old man who had previous pars plana vitrectomy and transscleral suture fixation of a posterior chamber (PC) IOL had lens subluxation caused by slippage of the haptic from a fixation suture. The IOL was dangling in the liquefied vitreous, preventing direct visualization of the displaced haptic. The displaced haptic was directly grasped from the opposite side with an intraocular forceps through a limbal wound and resutured to the sclera. Because only the end-grip intraocular forceps was required, this technique provides anterior segment surgeons an alternative technique of repositioning scleral-fixated PC IOLs.

Treatment of chronic pain by spinal cord stimulation.

Chronic pain after injury of the nervous system is difficult to treat. This report describes our experience in the treatment of chronic pain by spinal cord stimulation (SCS) in four patients (three men and one woman, with ages ranging from 61 to 73 yr). One patient had chronic intractable pain due to a spinal cord injury, one had post-herpetic neuralgia, one had failed back surgery syndrome, and one had brachial plexus and spinal cord injuries. A permanent spinal cord stimulator was implanted in the patient with spinal cord injuries. In the other three patients, a permanent spinal cord stimulator was implanted after a successful trial stimulation with temporarily implanted electrodes. After 19 to 25 months (mean, 21 mo) of follow-up, three patients had satisfactory improvement of pain and one patient had temporary pain relief, but pain recurred two months after implantation. One patient had recurrence of pain after migration of an electrode, and the pain-relieving effects of SCS returned after repositioning the electrode. According to this preliminary experience, SCS is effective for pain reduction in selected patients with chronic pain. Long-term follow-up is mandatory to evaluate the benefits and complications of SCS for relief of chronic pain.

Na,K-ATPase in lens epithelia from patients with senile cataracts.

Sodium-potassium-adenosine triphosphatase (Na,K-ATPase) has long been recognized for its role in regulating electrolyte concentrations in the lens, within which the electrolyte balance is vital to lens transparency. In this study, we compared the abundance of the alpha-subunit of Na,K-ATPase in lens epithelia of patients with senile cataracts, in order to examine the role of this enzyme in various types of lens opacity. Human lens epithelia were collected from 27 patients with senile cataracts who had undergone phacoemulsification. The type and the severity of lens opacity were graded and scored according to the Lens Opacities Classification System II. The mean age of the patients was 67.5 years (range, 46-80 yr). Abundance of the Na,K-ATPase alpha-subunit peptide in the lens epithelium was quantified by means of Western immunoblotting. Immunoblotting revealed that the amount of Na,K-ATPase alpha-subunit tended to decrease with increased cataract severity. In hypermature cataracts, the Na,K-ATPase alpha-subunit was barely detectable. The amount of alpha-subunit of Na,K-ATPase was inversely correlated with the overall severity of cataract (r = -0.64, p = 0.002). However, the inverse correlation was significant only in the cortical region (p = 0.027). As the cortex is located adjacent to the lens epithelium, it is directly affected by the loss of function of Na,K-ATPase in the epithelium. Such loss could result in water accumulation, vesicles, water clefts, Morgagnian globule formation, and Morgagnian cataract.

Traumatic wound dehiscence after penetrating keratoplasty: clinical features and outcome in 21 cases.

PURPOSE: To describe the characteristics, causes, treatment, and outcome, particularly the fate of the intraocular implant and visual acuity, of traumatic wound dehiscence occurring in patients who had penetrating keratoplasty (PK). METHODS: Between 1989 and 1997, 21 corneal transplant patients sustained traumatic wound dehiscence and were treated at our hospital. Graft dehiscence was managed with primary wound closure in all patients, except in some who required some combination of anterior vitrectomy, intraocular lens removal and reimplantation, and corneal regrafting. RESULTS: The incidence of traumatic wound dehiscence among patients on whom we performed PK over a period of 9 years was 2.53%. This complication occurred, on average, 3.4 years (2 months to 13 years) after PK. Most patients (76%) were men, and the average age at dehiscence was 59 years (range, 15-82 years). All corneal ruptures occurred at the graft-host junction, which had neither particular site preference nor quadrant specificity. Of the 16 eyes that were pseudophakic, nine (56.3%) had either loss of implants or dislocation of intraocular lens so severe that lens removal was imperative. In the end, only five (23.8%) of the 21 grafts retained clarity. Of the remaining 16 eyes, seven were regrafted, of which five (71.4%) grafts remained clear. Of the nine grafts that became opaque, six eyes (28.6%) atrophied. At the last follow-up, only six eyes (28.6%) had visual acuity of 20/200 or better, and six eyes had no perception of light. Final visual acuity was found to correlate inversely with the severity and extent of wound separation. CONCLUSION: Traumatic wound dehiscence is not rare after PK. The elderly corneal transplant patient may be more prone to such an injury. Corneal rupture at the graft-host junction in all of our cases means the persistence of wound weakness after PK. Although graft survival and visual outcome are generally poor after the injury, the restoration of a satisfactory visual result is possible after regrafting, insofar as the involved eye is free of intractable glaucoma or posterior segment damage.

Seborrheic keratosis of conjunctiva simulating a malignant melanoma: an immunocytochemical study with impression cytology.

OBJECTIVE: Seborrheic keratosis on the conjunctiva appears to have never been reported in the literature. The authors report here a well-documented case of seborrheic keratosis of conjunctiva clinically simulating a malignant melanoma. DESIGN: Case report. METHODS: A 66-year-old man presented with a juxtalimbal pigmented tumor involving the temporal conjunctiva of his left eye. Because of the rapid enlargement of the mass within a period of 5 months, a clinical diagnosis of malignant melanoma was made. Cytopathologic examinations were performed by impression cytology before the patient underwent a wide en-block excision of the tumor. MAIN OUTCOME MEASURES: Cytologic features were studied by impression cytology with periodic acid-Schiff-Papanicolaou stain. Immunochemical characteristics of tumor cells were studied by immunochemical stain of cytokeratin and HMB-45. Tumor morphology was observed by histopathologic examination. RESULTS: Impression cytology disclosed basaloid cells intermixing with squamoid cells, and these cells demonstrated positive immunoreactivity to cytokeratin and no reactivity to HMB-45. Histopathologic examination of the tumor specimen established the diagnosis of seborrheic keratosis, and the results of immunohistochemical staining were consistent with those of the impression cytology with immunocytochemical staining. CONCLUSION: The authors describe the first case report of conjunctival seborrheic keratosis and present its immunocytochemical and immunohistochemical characteristics. Such a benign lesion can clinically mimic a malignant melanoma.

Regression of orthotopic brain tumors by cytokine-assisted tumor vaccines primed in the brain.

This study investigated the therapeutic effects of a rat glioma cell line, C6, that was engineered to secrete mouse GM-CSF (mGM-CSF) on intracerebral (i.c.) brain tumors. Significant antitumor immunity was induced in rats when the live or irradiated mGM-CSF-secreting tumor vaccine was implanted i.c. The antitumor activity was effective on small tumors and, to a lesser extent, on large tumors or tumors existing in vivo for a longer duration. Immunohistochemical analysis revealed cellular infiltrates (granulocytes, macrophages, and CD4+ and CD8+ T cells) at both the vaccine site and the tumor site, indicating that immune responses were similarly activated when tumor vaccine was inoculated in the brain, as at the subcutis. Additional studies demonstrated that the therapeutic effects of tumor vaccines on the large tumors or the long-existing tumors were enhanced by strategies such as increasing the dosage of tumor vaccines, using combined vaccines consisting of mGM-CSF and human interleukin-2, or combining tumor vaccine with herpes simplex virus thymidine kinase/ganciclovir treatment. All of the modified strategies yielded synergistic therapeutic effects on the large tumor burdens. The data presented herein suggest that cytokine gene therapy is highly promising for the treatment of i.c. gliomas.

Transscleral fixation of foldable intraocular lens after pars plana lensectomy in eyes with a subluxated lens.

Two patients had a subluxated crystalline lens: one with Marfan's syndrome and the other with traumatic cataract. A pars plana lensectomy with transscleral fixation of a foldable silicone intraocular lens rather than a conventional poly(methyl methacrylate) lens was performed. Excellent, rapid visual rehabilitation was obtained using this small incision technique.

Efficacy of media enriched with nonlactate-generating substrate for organ preservation: in vitro and clinical studies using the cornea model.

BACKGROUND: Using a rabbit cornea model, our recent study demonstrated that Chen Medium (CM), an isotonic media enriched with nonlactate-generating high-energy substrates, is very effective for organ preservation. In the present study, the efficacy of CM is further evaluated with human corneas METHODS: The effectiveness of CM and Optisol for preserving the endothelial integrity of human corneas in vitro was evaluated by scanning electron microscopy. Clinical efficacy was evaluated in a total of 83 patients: 10 patients with keratoconus grafted randomly with either CM- or Optisol-stored cornea of the same donor, and 73 patients with various conditions grafted with CM-stored corneas. After surgery, visual acuity and quality of the graft were monitored for up to 4.6 years. RESULTS: The scanning electron microscopic study revealed that after 11-day storage at 4 degrees C, the CM-stored cornea had only marginal disruptive changes, 9.4+/-1.1%, in endothelial cells, as opposed to 42.4+/-4.6% of the Optisol-stored cornea. All 78 CM-stored corneas, including 67 with 12.2- to 17.7-hr death-to-storage time, 3-7.6 days of storage time, and initial marginal quality before storage, were successfully transplanted. These grafts were thin and clear, with an excellent epithelial integrity and without significant changes in endothelial cell density. Five Optisol-stored corneas were also successfully grafted; one of them, however, was edematous for about 4 weeks, and all the grafts were slightly thicker with substantial endothelial cell loss. CONCLUSION: Using a cornea model, present and recent studies show that CM is very effective for preserving tissue viability and endothelial integrity. Previous study revealed that CM-stored tissues maintained high levels of ATP and metabolic function, with suppression of lactate formation and accumulation. Thus, these findings support the concept that preservation of tissue viability is closely associated with the ability of the tissues to retain metabolic activity, to generate ATP efficiently, and to prevent acidosis effectively during storage.