Capsular Sites Research Articles

Abstract 632: Establishment and characterization of xenografts and cell lines trom nasopharyngeal carcinoma

Abstract Nasopharyngeal carcinoma (NPC) is a common cancer among ethnic Cantonese living in Hong Kong and southern China. It is closely associated with Epstein-Barr virus (EBV) infection. Unfortunately, there are very few representative xenografts and cell lines established from NPC available for investigation. Most of the NPC xenografts established have been passaged in immune deficient animals for over 20 years and may not be representative of the original NPC in patients. For in vitro NPC cell lines, there is only one cell line which retains the EBV genomes. Other NPC cell lines have all lost their EBV episomes. Furthermore, many of these NPC cell lines are found to be contaminated with genetic components from HeLa cells (HPV16 genome) which raised issues on their origins and limited their uses as NPC cells. There is great urgency in establishment new NPC cell lines both in vivo and in vitro for various research investigations and for the study of pathogenic role of EBV infection in NPC. We have carried out continuous efforts since 2010 to attempt establishment of new NPC xenografts and cell lines from surgical and biopsies NPC tissues. NPC tissues from resected recurrent NPC and biopsies from primary NPC were explanted to subrenal capsular sites and maintained for four months to one year to observe for growth of new NPC xenografts. Attempts to establish new NPC celles were also carried out. At present, we have successfully established 3 NPC xenografts (Xeno 23, Xeno 32, Xeno 47) which could be passages at subcutaneously sites and one in vitro NPC cells (NPC43) which harbors EBV genomes. The growth properties and genetic alterations of these newly established NPC cell lines have been characterized. Novel genetic alterations and growth signaling pathways were observed in these newly established NPC cell lines and may represent driver mutations and signaling pathways essential for progression of NPC. Profiling of EBV gene expression of these newly established NPC cell lines revealed predominant latent EBV infection and expression of EBV-encoded mRNA from the BART transcripts which are characteristic of EBV infection in NPC. The establishment of new NPC cells will contribute to investigate the pathogenesis of NPC and its intimate relationship with EBV infection. Funding acknowledgement: University Grant Council (HK) grants (AoE NPC (AoE/M-06/08); TBRS (T12-401/13R); GRF); University of Hong Kong (CRCG and SRT cancer) Citation Format: WT Lin, L Xia, Dan Dan Wen, CM Tsang, KW Lo, ML Lung, George Sai-Wah Tsao. Establishment and characterization of xenografts and cell lines trom nasopharyngeal carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 632.

A Tissue Graft Model of DNA Damage Response in the Normal and Malignant Human Prostate

DNA damage responses are relevant to prostate cancer initiation, progression and treatment. Few models of the normal and malignant human prostate that maintain stromal-epithelial interactions in vivo exist in which to study DNA damage responses. We evaluated the feasibility of maintaining tissue slice grafts at subcutaneous vs subrenal capsular sites in RAG2(-/-)γC(-/-) mice to study the DNA damage responses of normal and malignant glands. We compared the take rate and histology of tissue slice grafts from fresh, precision cut surgical specimens that were maintained for 1 to 4 weeks in subcutaneous vs subrenal capsular sites. Induction of γH2AX, p53, ATM and apoptosis was evaluated as a measure of the DNA damage response after irradiation. The take rate of subcutaneous tissue slice grafts was higher than typically reported but lower than at the subrenal capsular site. Subcutaneous tissue slice grafts frequently showed basal cell hyperplasia, squamous metaplasia and cystic atrophy, and cancer did not survive. In contrast, normal and malignant histology was well maintained in subrenal capsular tissue slice grafts. Regardless of implantation site the induction of γH2AX and ATM occurred in tissue slice graft epithelium 1 hour after irradiation and decreased to basal level by 24 hours, indicating DNA damage recognition and repair. As observed previously in prostatic ex vivo models, p53 was not activated. Notably, tumor but not normal cells responded to irradiation by undergoing apoptosis. To our knowledge this is the first study of DNA damage responses in a patient derived prostate tissue graft model. The subrenal capsular site of RAG2(-/-)γC(-/-) mice optimally maintains normal and malignant histology and function, permitting novel studies of DNA damage responses in a physiological context.

High frequency of capsular knee involvement in polymyalgia rheumatica/giant cell arteritis patients studied by positron emission tomography

Peripheral arthritis has been described in up to 50% of PMR patients, with knee involvement in the majority. This study was designed to evaluate by PET/CT the knees of patients with PMR and GCA and to identify the knee structures involved by inflammation. Twenty-five consecutive patients with PMR (19) or GCA (6) were studied in comparison with 25 age- and sex-matched controls who underwent PET/CT for initial staging of cancer. Clinical features, ESR and CRP were evaluated. Simultaneous FDG-PET and CT imaging from the skull base to the knee was performed after injection of 4.8-5.2 MBq of [(18)F]FDG per kilogram body weight. The knee anatomical structures being evaluated included joints, fibrous capsule, synovial recesses and bursae. At PET/CT, 21/25 patients (84%) showed bilateral diffuse uptake at the knees. The tracer clearly outlined the contour of the fibrous capsule. In 50 knees, 90% of capsular sites were involved by inflammation in comparison with 23% of intracapsular sites and 4.7% of extracapsular sites (P < 0.0001). No correlation was found between PET/CT results and ESR or CRP. FDG uptake, with a pattern similar to that observed in 96% of PMR/GCA patients, was seen in 20% of controls (P = 0.03). Our findings suggest that bilateral capsulitis of the knee is detectable in most PMR/GCA patients if a sensitive imaging technique such as PET/CT is used.

MRI Appearance of the Pectinofoveal Fold

The pectinofoveal fold is an intraarticular structure of the hip that has had only limited study in the clinical and anatomic literature. This fold may resemble a hip plica; however, symptomatic hip plicae are now being recognized and treated at hip arthroscopy. We wished to determine the frequency and appearance of the pectinofoveal fold on hip MR arthrography. By defining the variations in its appearance, the normal pectinofoveal fold can be distinguished from pathologic hip plicae. One hundred fifty-two hip MR arthrography examinations of patients who subsequently underwent hip arthroscopy were retrospectively reviewed. Each MR examination was reviewed for the presence of a pectinofoveal fold. If present, the fold was measured in the anteroposterior, mediolateral, and superior-inferior dimensions; evaluated for smooth or irregular contour; and evaluated for a femoral or capsular site of insertion. The pectinofoveal fold was visualized on hip MR arthrograms in 144 of the 152 (95%) patients and visualized at hip arthroscopy in 150 of the 152 (99%) patients. The average thickness of the fold was 2.6 mm (range, 1-13 mm) in the mediolateral dimension and 17 mm (range, 1-32 mm) in the anteroposterior dimension. The average length of the fold in the superior-inferior dimension was 23.3 mm (range, 7-44 mm). The pectinofoveal fold had a smooth contour in 75 of the 144 (52%) patients with examinations that showed the fold and an irregular contour in 69 of 144 (48%) patients. The fold was found to insert onto the capsule in 108 of 144 (75%) patients and onto the femur in the remaining 36. The pectinofoveal fold should almost always be visualized at MR arthrography. The fold can have various appearances and attachment sites, and these normal variations should not be mistaken for fold abnormalities. These findings should be useful in distinguishing this normal structure from normal and pathologic plicae.

Organogenesis of Pancreatic Anlagen Allografted in Rats

Aims To study the possibility of revascularization, growth, and differentiation of embryonic pancreatic anlagen transplanted to adult hosts. While transplantations of pancreas and islets are the main methods to cure diabetes mellitus, the donor source is in shortage. So it’s necessary to find a new source for transplantation. Methods The pancreas from embryonic day 14.5 (E14.5) and 15.5 (E15.5) Lewis rat embryos were implanted into either intraperitoneal or subrenal capsular site of healthy Lewis rats. at 3 weeks or 6 weeks after implantation, the pancreatic anlagen in the host rats were resected for size measurements, as well as histopathologic and immunohistochemical examinations. Results Three weeks after implantation into the renal-capsular site, the size of both E14.5 and E15.5 pancreatic anlagen had enlarged 10- to 15-fold with differentiation of acinar components upon histological examination. Moreover, increasing numbers of β cells and islets stained positive for insulin, and newly generated vessels were observed around the tissues. Continued proliferation of the endocrine islets in E14.5 pancreatic anlagen grafts was observed after another 3 weeks, whereas further proliferation in the E15.5 pancreatic anlagen graft was not seen. Additionally fibrosis appeared in the exocrine component of both E14.5 and E15.5 pancreatic anlagen at this time point. When implanted into intraperitoneal site, enlarged E15.5 pancreatic anlagen with proliferatels β cells were also observed after 3 weeks. However, both the size of the pancreatic anlagen and the proliferation of the β cells were much less than that in the subrenal capsular site. Conclusions The allografted E14.5 and E15.5 pancreatic anlagen revascularised and grew into tissues that were structurally similar to normal mature rats pancreatic tissue. Adequate embryonic age for the transplantation of pancreatic anlagen is 14.5 and 15.5 days old. Subrenal capsula is a more suitable site than the peritoneal cavity for implantation of pancreatic anlagen.

Establishment and characterization of a new human renal cell carcinoma cell line (KG-2) and metastatic model in nude mice

A new cultured cell line (KG-2) derived from human renal cell carcinoma and a metastatic model in nude mice were studied. KG-2 was cultured from renal cell carcinoma (clear cell carcinoma) of the left kidney. In vitro doubling time of KG-2 was approximately 50 hours. KG-2 cells produced tumors in both the subcutaneous and renal sub-capsular space in nude mice, with tumorigenicity of 75%, showing no difference between the two sites. Histologically, tumors formed in the subcutaneous sites were hypovascular granular cell carcinoma. Moreover, each tumor was encapsulated by a thick fibrous capsule and never produced distant metastasis or invasion into the surrounding tissue. However, tumors formed in the subrenal capsular space were clear cell carcinoma. These tumors were hypervascular, and produced distant metastases. The most common metastatic site was the lung. Immunohistochemical analysis using anti-human collagenase type IV antibody on tumors formed in subcutaneous and subrenal capsular sites demonstrated that the expression of this enzyme in tumors formed in the subrenal capsular space was much higher than that in tumors formed in the subcutaneous site. Additionally, immunohistochemical study using anti-mouse collagen type IV antibody, a major components of the vascular wall, demonstrated many small densely growing vessels in tumors formed in the subrenal capsular space. In contrast, few vessels were produced in tumors formed in subcutaneous sites. These findings suggest that factors relating to the different injection sites may regulate the production of collagenase type IV secreted by KG-2 cells and neovascularity in nude mice. This metastatic model may be useful in the study of the mechanism of cancer metastases.

Emotional facial paresis with striatocapsular infarction

Emotional facial paresis (EFP) refers to hemifacial paresis of emotionally evoked or spontaneous smiling or weeping with preserved volitional movements of the face. The anatomical location for this phenomenon has been controversial. We report a 15-year-old boy with EFP. Follow-up MRI demonstrated infarction limited to the head of the caudate, putamen and anterior limb of the internal capsule on the contralateral side. This case lends support for selective damage at a striatal or capsular site in the production of EFP.

Mechanical properties of the dog renal capsule.

The renal capsule is an important determinant of whole kidney volume/pressure relationships. To gain further insights into its possible role we examined the mechanical properties of the dog renal capsule using standard materials testing procedures. From each of four locations on the kidney surface, the following mechanical properties of the renal capsule were determined: elastic modulus (force/unit of cross-sectional area theoretically required to double the length of the specimen), tensile stiffness (force/unit width theoretically required to double the length of the specimen), ultimate strength (stress at time of fracture of the specimen), and maximum strain (percent strain at time of the fracture of the specimen). We found that the elastic modulus of the renal capsule from all capsular sites was substantially greater than values previously reported for dog aorta. The stiffness of the capsule covering the anterior-posterior surface of the kidney was found to be about 50% greater than the stiffness of the capsule covering the lateral and polar surfaces of the kidney. The ultimate strength of the anterior-posterior capsule was significantly greater than that of the lateral or polar capsule. This finding may explain the clinical observation that the spontaneous rupture of the renal capsule and parenchyma associated with the acute swelling of transplant rejection is confined almost exclusively to the lateral and polar portions of the renal capsule and cortex. The mean maximum strain at each capsular site was about 35%. This degree of circumferential expansion corresponds to about a doubling of kidney volume. Thus, this observation suggests that the renal capsule is at risk to undergo spontaneous rupture when renal volume increases of this magnitude are observed.

Electrophysiological studies of some deep cerebral structures in man

In stereotactic operations for Parkinson's disease and other disturbances of voluntary movement, the electrophysiological activity of the brain was recorded along the trajectory of the coagulating electrode. The method permitted a clear distinction between white matter and gray matter. Within the thalamus and elsewhere it was possible to differentiate functionally the activity of numerous nuclei (pulvinar, lateralis posterior, ventrolateralis, ventralis posterior medialis and lateralis, globus pallidus, amygdala). Somato-topically organized potentials were recorded in the nucleus ventralis posterior upon natural peripheral stimulation. In several nuclei it was possible to observe rhythmical activities occurring at the same frequency as the muscle tremor. These activities must have been the result of the muscle tremor in some thalamic structures (ventralis posterior lateralis) but they appear to be more directly related to the ‘generation’ of the tremor in other nuclei (lateralis posterior and possibly ventralis lateralis). Stimulation of thalamic and capsular sites produced sensory or motor effects fully in agreement with the results of derivation from these points. This method made it possible to improve the accuracy of the placement of the lesion and to demonstrate again the considerable anatomical variability of the individual thalamus.