Characterizing the Robustness of Distinct Clinical Assessments in Identifying Dry Eye Condition of Animal Models.

PURPOSE: The study aims to characterize the robustness of distinct clinical assessments in identifying the underlying conditions of dry eye disease (DED), with a specific emphasis on the involvement of conjunctival goblet cells. METHODS: Seven rabbits receiving surgical removal of the lacrimal and Harderian glands were divided into two groups, one with ablation of conjunctival goblet cells by topical soaking of trichloroacetic acid (TCA) to the bulbar conjunctiva (n = 3) and one without (n = 4), and the conditions of DED were assessed weekly using Schirmer test, tear breakup time (TBUT), tear osmolarity, and National Eye Institute (NEI) fluorescein staining grading. After 8 weeks, the rabbits were sacrificed, and the eyes were enucleated for histopathological examination. RESULTS: Histopathological analysis revealed corneal epithelial thinning in both groups. While TCA soaking significantly decreased the density of conjunctival goblet cells, DED rabbits without TCA also showed a partial reduction in goblet cell density, potentially attributable to dacryoadenectomy. Both groups showed significant decreases in Schirmer test and TBUT, as well as an increase in tear osmolarity. In DED rabbits with TCA soaking, tear osmolarity increased markedly, suggesting that tear osmolarity is highly sensitive to loss and/or dysfunction of conjunctival goblet cells. Fluorescein staining was gradually and similarly increased in both groups, suggesting that fluorescein staining may not reveal an early disruption of the tear film until the prolonged progression of DED. CONCLUSION: The Schirmer test, TBUT, tear osmolarity, and NEI fluorescein grading are distinct, yet complementary, clinical assessments for the evaluation of DED. By performing these assessments in definitive DED rabbit models, both with and without ablation of conjunctival goblet cells, the role of these cells in the homeostasis of tear osmolarity is highlighted. Characterizing the robustness of these assessments in identifying the underlying conditions of DED will guide a more appropriate management for patients with DED.

A modified SELEX approach to identify DNA aptamers with binding specificity to the major histocompatibility complex presenting ovalbumin model antigen.

Aptamers have sparked significant interest in cell recognition because of their superior binding specificity and biocompatibility. Cell recognition can be mediated by targeting the major histocompatibility complex (MHC) that presents short peptides derived from intracellular antigens. Although numerous antibodies have demonstrated a specific affinity for the peptide-MHC complex, the number of aptamers that exhibit comparable characteristics is limited. Aptamers are usually selected from large libraries via the Systemic Evolution of Ligands by Exponential Enrichment (SELEX), an iterative process of selection and PCR amplification to enrich a pool of aptamers with high affinity. However, the success rate of aptamer identification is low, possibly due to the presence of complementary sequences or sequences rich in guanine and cytosine that are less accessible for primers. Here, we modified SELEX by employing systemic consecutive selections with minimal PCR amplification. We also modified the analysis by selecting aptamers that were identified in multiple selection rounds rather than those that are highly enriched. Using this approach, we were able to identify two aptamers with binding specificity to cells expressing the ovalbumin alloantigen as a proof of concept. These two aptamers were also discovered among the top 150 abundant candidates, despite not being highly enriched, by performing conventional SELEX. Additionally, we found that highly enriched aptamers tend to contain fractions of the primer sequence and have minimal target affinity. Candidate aptamers are easily missed in the conventional SELEX process. Therefore, our modification for SELEX may facilitate the identification of aptamers for more application in diverse biomedical fields. Significance: we modify the conventional method to improve the efficiency in the identification of the aptamer, a single strand of nucleic acid with binding specificity to the target molecule, showing as a proof of concept that this approach is particularly useful to select aptamers that can selectively bind to cells presenting a particular peptide by the major histocompatibility complex (MHC) on the cell surface. Given that cancer cells may express mutant peptide-MHC complexes that are distinct from those expressed by normal cells, this study sheds light on the potential application of aptamers to cancer cell targeting.

Cellular differentiation into hyphae and spores in halophilic archaea.

Several groups of bacteria have complex life cycles involving cellular differentiation and multicellular structures. For example, actinobacteria of the genus Streptomyces form multicellular vegetative hyphae, aerial hyphae, and spores. However, similar life cycles have not yet been described for archaea. Here, we show that several haloarchaea of the family Halobacteriaceae display a life cycle resembling that of Streptomyces bacteria. Strain YIM 93972 (isolated from a salt marsh) undergoes cellular differentiation into mycelia and spores. Other closely related strains are also able to form mycelia, and comparative genomic analyses point to gene signatures (apparent gain or loss of certain genes) that are shared by members of this clade within the Halobacteriaceae. Genomic, transcriptomic and proteomic analyses of non-differentiating mutants suggest that a Cdc48-family ATPase might be involved in cellular differentiation in strain YIM 93972. Additionally, a gene encoding a putative oligopeptide transporter from YIM 93972 can restore the ability to form hyphae in a Streptomyces coelicolor mutant that carries a deletion in a homologous gene cluster (bldKA-bldKE), suggesting functional equivalence. We propose strain YIM 93972 as representative of a new species in a new genus within the family Halobacteriaceae, for which the name Actinoarchaeum halophilum gen. nov., sp. nov. is herewith proposed. Our demonstration of a complex life cycle in a group of haloarchaea adds a new dimension to our understanding of the biological diversity and environmental adaptation of archaea.

Chest X-ray and CT findings of early H7N9 avian influenza cases.

BACKGROUND: The H7N9 strain of bird flu is a new type of avian flu that was identified at the end of March 2013. The disease is concerning because most patients have become severely ill. PURPOSE: To study the X-ray and computed tomography (CT) findings of early H7N9 avian influenza cases. MATERIAL AND METHODS: Chest radiography and CT were performed in six patients with H7N9 avian influenza within 1-20 days after onset. The CT examinations included conventional spiral CT and high-resolution CT. The findings on the radiography and CT images were analyzed. RESULTS: Abnormal X-ray and CT findings were present in all of the patients. All of the cases had acute onset. In the early stage, the right lung was more commonly affected (particularly in the right upper and middle lobes). The lesions rapidly expanded to the entire lungs and were characterized primarily by ground-glass opacities (GGOs) combined with consolidation. Diffuse GGO was observed in all six cases (1 was symmetric, and 5 were non-symmetric). Local consolidation was found in four cases, and lobar consolidation was found in two cases. Normal lung tissue was observed between the lesions. Pleural thickening was common and was combined with pleural/pericardial effusion or mediastinal lymph node enlargement. Reticular changes, centrilobular nodules, and the tree-in-bud sign were observed in some cases, but reticular changes, bronchial wall thickening, and hyperinflation were not found. CONCLUSION: Radiological changes associated with both acute pneumonia and acute interstitial inflammation were observed in early H7N9 avian influenza cases. Serial chest X-rays were useful for the diagnosis and severity assessment of the disease. CT may provide a more accurate assessment of the lung pathology.

Pulmonary CT findings in relapsing polychondritis.

BACKGROUND: Relapsing polychondritis (RPC) is a rare rheumatic disease characterized by recurrent inflammation of cartilaginous structures, with airway involvement a major cause of morbidity and mortality. PURPOSE: To retrospectively evaluate airway and lung abnormalities in RPC with computed tomography (CT). MATERIAL AND METHODS: From January 2004 to May 2009, 21 patients with RPC (12 men, 9 women; 13-65 years old) underwent chest CT examinations. Two chest radiologists evaluated the CT images retrospectively. Abnormal findings, including airway stenosis, airway malacia, air trapping, and airway wall thickening with or without calcifications, were observed and noted. RESULTS: Major abnormal CT findings were observed in eight patients (38.1%), which included airway wall thickening (n=7), airway stenosis (n=6), airway malacia (n=6), airway wall calcification (n=8), and air trapping (n=3). Mediastinal lymph nodes were found in 12 patients. Lung infection was identified in four patients and interstitial lung disease in six patients. CONCLUSION: The CT findings in patients with RPC consisted mainly of airway wall thickening, airway stenosis, airway malacia, airway wall calcification, and air trapping.

Nonalcoholic fatty liver disease: quantitative assessment of liver fat content by computed tomography, magnetic resonance imaging and proton magnetic resonance spectroscopy.

OBJECTIVE: The purpose of this study was to evaluate the clinical application of imaging technology in the quantitative assessment of fatty liver with magnetic resonance imaging (MRI) and proton MR spectroscopy. METHODS: Overall 36 patients with diffuse fatty liver who had undertaken the computed tomography (CT) scan, MRI and proton MR spectroscopy (1H MRS) were analyzed. Their body mass index (BMI) was measured and their liver to spleen CT ratio (L/S) calculated on the plain CT scan. MR T1-weighted imaging (T1WI) was obtained with in-phase (IP) and out-of-phase (OP) images. T2-weighted imaging (T2WI) was acquired with or without the fat-suppression technique. The liver fat content (LFC) was quantified as the percentage of relative signal intensity loss on T1WI or T2WI images. The intrahepatic content of lipid (IHCL) was expressed as the percentage of peak value ratio of lipid to water by 1H MRS. RESULTS: The results of BMI measurement, CT L/S ratio, LFC calculated from MR T1WI and T2WI images, as well as IHCL measured by 1H MRS were 27.26 +/- 3.01 kg/m2, 0.88 +/- 0.26, 13.80 +/- 9.92%, 40.67 +/- 16.04% and 30.98 +/- 20.43%, respectively. The LFC calculated from MR T1WI, T2WI images and IHCL measured by 1H MRS correlated significantly with the CT L/S ratio (r = -0.830, P = 0.000; r = -0.736, P = 0.000; r = -0.461, P = 0.005, respectively). BMI correlated significantly only with the liver fat contents measured by T1WI IP/OP and 1H MRS (r = -0.347, P = 0.038; r = -0.374, P = 0.025, respectively). CONCLUSION: CT, MR imaging and 1H MRS were effective methods for the quantitative assessment of LFC. The MR imaging, especially 1H MRS, would be used more frequently in the clinical evaluation of fatty liver and (1)H MRS could more accurately reflect the severity of fatty liver.