Variability in the Epithelial Expression of Ki67 and Polarization Characteristics of Collagen in Odontogenic Cysts and Tumours

ABSTRACT Objective: To assess the proliferation of epithelium (using the Ki67 index) and the polarization pattern of collagen in selected odontogenic cysts and tumours. In addition, an exploratory analysis of the effect of inflammation on the proliferation rate was done. Material and Methods: Following immunohistochemical staining, the labelling/proliferation index of Ki67 was calculated. The thickness and corresponding polarization colour of 100 juxta-epithelial picrosirius red-stained collagen fibers were assessed using linear micrometry with an eyepiece reticule under × 1000 magnification. Inflammation was graded subjectively as mild, moderate, and severe. Results: Overall Ki-67 expression was higher in the radicular cyst, Odontogenic Keratocyst, Ameloblastoma, while suprabasal Ki-67 positivity was maximum in Odontogenic Keratocyst. The stromal collagen fibers in Ameloblastoma showed predominantly green birefringence, whereas Odontogenic Keratocyst had orange birefringence. There was no significant association of inflammation with Ki-67 expression or birefringence patterns. Conclusion: The highest Ki67 expression in the radicular cyst, followed by Odontogenic Keratocyst and Ameloblastoma. Differences in the collagen maturation pattern were noted innately in five lesions studied and were further influenced by inflammatory changes. Epithelial proliferation and concomitant expression of thickness and maturity of the stromal collagen are innate features of the lesion further influenced by inflammation in various odontogenic cysts and tumours and may, in turn, guide the clinical behavior.

Análise do processo de reparo alveolar em camundongos fêmeas senescentes 129/Sv-WT e 129/Sv 5-LOKO sob terapia com bifosfonato de alta potência / Alveolar bone repair of 129/Sv WT and 129/Sv 5LOKO female senescent mice under therapy with high potency bisphosphonate

O processo de senescência acarreta uma série de modificações fisiológicas com declínio das funções das atividades celulares e sistêmicas que se manifestam de maneira mais importante na população feminina pelo evento da menopausa, como a osteoporose. A fim de se minimizar tais efeitos, há a possibilidade de se utilizar medicamentos que diminuem o processo de remodelação óssea como os bifosfonatos nitrogenados (BF). Entretanto, o uso dessas drogas está intimamente relacionado ao desenvolvimento de osteonecrose dos maxilares (OM), principalmente quando associado a outros fatores de risco como as cirurgias bucais. Sabe-se que fisiologicamente a dinâmica do tecido ósseo depende também de eicosanóides derivados do metabolismo do ácido araquidônico (AA), como as enzimas cicloxigenase (COX) e 5 lipoxigenase (5LO). Deste modo, o objetivo do presente trabalho foi analisar o efeito BF ácido zoledrônico (ZL) e sua relação com o desenvolvimento da OM em camundongos fêmeas senescentes 129/Sv com e sem modificação genética para a enzima 5LO. Para tanto, foram utilizados 40 camundongos fêmeas senescentes 129/Sv, sendo 20 WT e 20 com alteração no gene 5LO (129 Alox5tm1Fun/J) (5LOKO), divididas em grupos: WT, tratadas com 0,01 ml de solução salina 0,9% estéril (SS) via intraperitoneal (IP) e ZL, tratadas com 250µg/Kg de ácido zoledrônico (ZL) IP diluído em solução salina estéril, ambas administradas 1 vez/semana por 7 semanas. Os grupos foram compostos por 5 animais cada (WT Controle ­ 7 e 21dias, WT ZL ­ 7 e 21 dias, 5LOKO Controle ­ 7 e 21 dias, 5LOKO ZL ­ 7 e 21dias), sendo as maxilas coletadas para análises em microCT, histopatológica, birrefringência, técnica imunohistoquímica e histomorfométricas. De modo geral, a microCT revelou deficiência significativa na microarquitetura óssea nos animais WT ZL em comparação com os demais. Do mesmo modo, a partir da análise histopatológica e de birrefringência da matriz colagenosa, observou-se padrão compatível com o desenvolvimento de OM no grupo WT ZL, com presença de infiltrado inflamatório intenso, atraso na neoformação óssea, presença de fraturas patológicas, e deficiência da matriz colagenosa e de células Runx-2+, TRAP+ e F4/80+. Os animais 5LOKO ZL apresentaram alterações compatíveis com atraso no processo de reparo especialmente no período de 7 dias, com menor quantidade de células Runx-2+ em comparação com o grupo 5LOKO Controle e pela qualidade da matriz óssea colagenosa com menor quantidade de fibras do espectro vermelho neste período, se igualando, porém, aos 21 dias. Deste modo, concluiu-se que o processo de reparo em camundongos fêmeas senescentes da linhagem 129/Sv WT e 5LOKO associados ao uso do BF ZL ocorreu de modo distinto, levando a quadro de OM nos animais WT e atraso nos animais 5LOKO, sem sinais histopatológicos que caracterizassem a doença. Deste modo, a inibição da enzima 5LO parece influenciar de maneira positiva o processo de reparo ósseo intramembranoso alveolar, mesmo na presença de fenótipo esqueletal osteopetrótico, sugerindo outros fatores relacionados à droga que favoreçam o desenvolvimento da OM no presente modelo animal(AU)

Senescence brings a number of physiological modifications with the decrease of cell and systemic activities and function that manifest in an important way in female population due to the event of menopause, as osteoporosis. In order to diminish these effects, there is the possibility of taking medication that decrease bone remodeling process, as the bisphosphonates containing nitrogen (BF). However, the use of these drugs is intimate related with the development of the osteonecrosis of the jaws (ON), especially when associated to other risk factors as oral surgery. It is known that physiologically, the dynamics of bone tissue also depends on the eicosanoids derivate from the arachidonic acid metabolism (AA), such as cyclooxygenase (COX) and 5 lipoxygenase (5LO) enzymes. In this way, the aim of the present study was to analyze the effects of the BF zoledrônico acid (ZL) and its relation with de development of ON in 129/SV old female mice with or without genetic modification for 5LO. Forty animals, 20 WT and 20 with 5LO gene alteration (129 Alox5tm1Fun/J) (5LOKO) were divided in groups: WT, treated with 0.01 ml of sterile 0.9% saline solution (SS) intraperitoneal (IP), and ZL, treated with 250µg/Kg of ZL IP diluted in SS, both administered once a week for 7 weeks. Groups contained 5 animals each (WT Control ­ 7 and 21 days, WT ZL ,7 and 21 days, 5LOKO Control, 7 and 21 days, and 5LOKO ZL, 7 and 21 days), and the maxillae removed for microCT, histopathology, birefringence, immunohistochemistry, and histomorphometric analysis. In general, microCT revealed significant deficiency in bone microarchitecture in WT ZL group in comparison to the other groups. In the same way, histopathological and birefringence analysis revealed histological pattern compatible with ON development in WT ZL group, presenting intense inflammatory infiltrate, late new bone formation, presence of pathological fractures, and deficiency in collagenous matrix, and also in Runx-2+, TRAP+, and F4/80. 5LOKO ZL animals presented alterations compatible with a late bone repair, especially at day 7, with decreased number of Runx-2+ cells in comparison to 5LOKO Control, and by the quality of collagenous bone matrix with decreased number of red spectra fibers in this period, however, being similar at day 21. From this, it could be concluded that alveolar bone repair of 129/SV WT and 5LOKO old female mice associated with the administration of ZL occurred in different ways, leading to a picture of ON in the WT animals, and late bone repair in the 5LOKO animals, without histopathological signs that could characterize the disease. In this way, inhibition of 5LO seems to influence intramembranous alveolar bone repair in a positive way, even in the presence of osteopetrotic skeletal phenotype, suggesting other factors related to the drug that favors the development of the ON in the present animal model(AU)

Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

Abstract: The special picrosirius red staining highlights the natural birefringence of collagen fibers when exposed to polarized light. The results from birefringence allow to evaluate the organization of the collagen fibers in the tissues. The authors intend to elucidate all steps to obtain and capture images of histological sections stained with picrosirius red and evaluated under polarized light microscopy, as well as possible artefacts that may occur.

An Immunohistochemical and Polarizing Microscopic Study of the Tumor Microenvironment in Varying Grades of Oral Squamous Cell Carcinoma

BACKGROUND: Invasion of epithelial cells into the connective tissue brings about massive morphological and architectural changes in the underlying stroma. Myofibroblasts reorganize the stroma to facilitate the movement of tumor cells leading to metastasis. The aim of this study was to determine the number and pattern of distribution of myofibroblasts and the qualitative and quantitative change that they cause in the collagen present in the stroma in various grades of oral squamous cell carcinoma (OSCC). METHODS: The study was divided into two groups with group I (test group, 65 cases) consisting of 29 cases of well-differentiated squamous cell carcinoma, 25 moderately differentiated SCC, and 11 poorly differentiated SCC, and group II (control group) consisting of 11 cases of normal mucosa. Sections from each sample were stained with anti-α-smooth muscle actin (α-SMA) antibodies, hematoxylin and eosin, and Picrosirius red. Several additional sections from each grade of OSCC were stained with Masson's trichrome to observe the changes in collagen. For the statistical analysis, Fisher's exact test, Tukey's post hoc honest significant difference test, ANOVA, and the chi-square test were used, and p < .05 was considered statistically significant. RESULTS: As the tumor stage progressed, an increase in the intensity α-SMA expression was seen, and the network pattern dominated in more dedifferentiated carcinomas. The collagen fibers became thin, loosely packed, and haphazardly aligned with progressing cancer. Additionally, the mean area fraction decreased, and the fibers attained a greenish yellow hue and a weak birefringence when observed using polarizing light microscopy. CONCLUSIONS: Myofibroblasts bring about numerous changes in collagen. As cancer progresses, there isincrease in pathological collagen,which enhances the movement of cells within the stroma.

Asymptomatic Localized Gastric Amyloidosis with Two Separate Lesions

Amyloidosis is a disease in which amyloid is abnormally accumulated in the tissue. The kidney and heart are the most commonly involved organs. Gastric involvement is relatively common in systemic disease; however, localized gastric amyloidosis is rare. Here, we report a case of localized gastric amyloidosis with two separate lesions. A 56-year-old woman underwent a health surveillance checkup without any noticeable symptoms. She was under medication for diabetes and dyslipidemia, and was otherwise healthy. On surveillance upper endoscopy, an irregularly shaped hyperemic elevated erosion at the gastric fundus and a 1.5-cm, yellowish subepithelial tumor-like lesion with intact overlying mucosa at the lesser curvature of the lower body of the stomach were detected. Endoscopic biopsy revealed submucosal eosinophilic material deposition in both lesions. Congo-red staining showed amyloid deposit appearing as a yellow-green birefringence under polarizing microscopy. Echocardiography, abdominal sonography, and colonoscopy revealed no abnormality. The patient was diagnosed as having localized gastric amyloidosis and is now undergoing regular follow-up without any treatment. Localized gastric amyloidosis is a rare disease that may mimic nonspecific gastritis or subepithelial tumor. However, endoscopic biopsy with appropriate staining may be diagnostic and thorough evaluation for systemic involvement is important.

Localized Gastric Amyloidosis with Kappa and Lambda Light Chain Co-Expression

Esophagogastroduodenoscopy for cancer screening was performed in a 55-year-old woman as part of a health screening program, and revealed a depressed lesion approximately 20 mm in diameter in the lesser curvature of the mid-gastric body. Several biopsy specimens were collected as the lesion resembled early gastric cancer; however, histopathologic evaluation revealed chronic active gastritis with an ulcer and amorphous eosinophilic material deposition. Congo red staining identified amyloid proteins, and apple-green birefringence was shown using polarized light microscopy. Immunohistochemical staining revealed the presence of kappa and lambda chain-positive plasma cells. There was no evidence of underlying plasma cell dyscrasia or amyloid deposition in other segments of the gastrointestinal tract. Echocardiography and computed tomography of the chest, abdomen, and pelvis did not show any significant findings. Thus, the patient was diagnosed with localized gastric amyloidosis with kappa and lambda light chain coexpression.

Relationship between meiotic spindles visualization and intracytoplasmic sperm injection outcomes in human oocytes / 대한해부학회지

In assisted reproductive techniques, the operator attempts to select morphologically best embryos to predict embryo viability. Development of polarized light microscope, which evaluates the oocytes' spindles according to birefringence of living cells, had been helpful in oocyte selection. The aim of this study is evaluating the relationship between meiotic spindles visualization and intracytoplasmic sperm injection (ICSI) outcomes in human oocytes. In this study, 264 oocytes from 24 patients with an average age of 30.5±7.5 years with infertility duration of 1 to 10 years were collected. The oocytes were randomly allocated to the control injection group (n=126) and the oocyte imaging group (spindle-aligned group) (n=138). In the spindle-aligned group, the meiotic spindle was identified by means of polarized light microscope to align the spindle at 6 or 12 o'clock. Then the spindle-aligned group was divided into three sub-groups based on spindle morphology: fine, average, and (poor). After ICSI, embryos were checked every 24 hours and scored; 72 hours later, high-grade embryos were transferred intravaginally to uterus. This study showed that the fertilization rate in the spindle-aligned group was higher than the control group (P<0.05). After cleavage, a positive correlation was observed between spindle morphology and embryo morphology. Among the sub-groups of spindle-aligned group, the embryos' morphology of the fine group was better than the other subgroups and embryos of the poor group had lower quality and more fragmentation. The results revealed that the selection of oocytes based on meiotic spindle imaging can significantly improve the rate of fertilization and embryo cleavage and certainly increase the rate of implantation.

Secondary Cutaneous Amyloidosis in a Patient with Mycosis Fungoides

Secondary cutaneous amyloidosis refers to clinically unapparent amyloid deposits within the skin in association with a pre-existing skin condition or skin tumors, such as basal cell carcinoma, porokeratosis, solar elastosis, Bowen's disease, and mycosis fungoides. A 70-year-old woman presented with a 6-month history of asymptomatic multiple yellowish plaques on both legs. She had been diagnosed with mycosis fungoides 7 years ago and was treated with psoralen and ultraviolet A radiation (PUVA) therapy, narrow-band ultraviolet B (UVB) therapy, and acitretin for 5 years. Finally, she reached complete remission of mycosis fungoides. However, new yellowish lesions started to appear 1 year after discontinuing the phototherapy. A physical examination revealed multiple yellowish plaques on both extremities. The plaques were well circumscribed and slightly elevated. All laboratory tests were normal. A biopsy specimen showed multiple nodular deposits of eosinophilic amorphous material in papillary dermis and upper reticular dermis. The deposits represented apple green birefringence on Congo red stain viewed under polarized light. Acellular small nodules in the upper dermis consisted of randomly oriented, non-branching, 6.67~12.7 nm thick amyloid fibrils on electron microscopy. We report an interesting and rare case of secondary cutaneous amyloidosis after narrow-band UVB therapy and PUVA therapy in a patient with mycosis fungoides.

Secondary Cutaneous Amyloidosis in a Patient with Mycosis Fungoides

Secondary cutaneous amyloidosis refers to clinically unapparent amyloid deposits within the skin in association with a pre-existing skin condition or skin tumors, such as basal cell carcinoma, porokeratosis, solar elastosis, Bowen's disease, and mycosis fungoides. A 70-year-old woman presented with a 6-month history of asymptomatic multiple yellowish plaques on both legs. She had been diagnosed with mycosis fungoides 7 years ago and was treated with psoralen and ultraviolet A radiation (PUVA) therapy, narrow-band ultraviolet B (UVB) therapy, and acitretin for 5 years. Finally, she reached complete remission of mycosis fungoides. However, new yellowish lesions started to appear 1 year after discontinuing the phototherapy. A physical examination revealed multiple yellowish plaques on both extremities. The plaques were well circumscribed and slightly elevated. All laboratory tests were normal. A biopsy specimen showed multiple nodular deposits of eosinophilic amorphous material in papillary dermis and upper reticular dermis. The deposits represented apple green birefringence on Congo red stain viewed under polarized light. Acellular small nodules in the upper dermis consisted of randomly oriented, non-branching, 6.67~12.7 nm thick amyloid fibrils on electron microscopy. We report an interesting and rare case of secondary cutaneous amyloidosis after narrow-band UVB therapy and PUVA therapy in a patient with mycosis fungoides.

Differential diagnosis of periapical cyst using collagen birefringence pattern of the cyst wall

OBJECTIVES: Periapical lesions, including periapical cyst (PC), periapical granuloma (PG), and periapical abscess (PA), are frequently affected by chemical/physical damage during root canal treatment or severe bacterial infection, and thus, the differential diagnosis of periapical lesions may be difficult due to the presence of severe inflammatory reaction. The aim of this study was to make differential diagnosis among PC, PG, and PA under polarizing microscope. MATERIALS AND METHODS: The collagen birefringence patterns of 319 cases of PC (n = 122), PG (n = 158), and PA (n = 39) obtained using a polarizing microscope were compared. In addition, 6 cases of periodontal fibroma (PF) were used as positive controls. RESULTS: Collagen birefringence was condensed with a thick, linear band-like pattern in PC, but was short and irregularly scattered in PG, and scarce or absent in PA. PF showed intense collagen birefringence with a short, palisading pattern but no continuous band-like pattern. The linear band-like birefringence in PC was ascribed to pre-existing expansile tensile stress of the cyst wall. CONCLUSIONS: In this study all PCs (n = 122) were distinguishable from PGs and PAs by their characteristic birefringence, despite the absence of lining epithelium (n = 20). Therefore, the authors suggest that the presence of linear band-like collagen birefringence of the cyst wall aids the diagnostic differentiation of PC from PG and PA.

Host inflammatory response to polypropylene implants: insights from a quantitative immunohistochemical and birefringence analysis in a rat subcutaneous model

ABSTRACT Objectives To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Materials and Methods Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. Results A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). Conclusions The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization.

Efeito da aplicação do ultrassom terapêutico durante 4 e 5 minutos por área do transdutor no processo de reparação de tendão de ratos / Effect of application times 3, 4 and 5 minutes ERA of therapeutic ultrasound in tendon injury of calcaneal rats

O objetivo deste estudo foi avaliar o efeito dos tempos de aplicação 3, 4 e 5 minutos por ERA do ultrassom terapêutico (UST) na organização das fibras de colágeno em lesão do tendão do calcâneo de ratos. Foram utilizados quarenta ratos machos Wistar, dos quais 32 sofreram tenotomia total do tendão do calcâneo e foram divididos em 5 grupos: GC, sem tenotomia e tratamento; GT, com tenotomia e sem tratamento; UST3, UST4 e UST5 submetidos à tenotomia e tratados com UST nos tempos de 3, 4 e 5 minutos por área de radiação efetiva respectivamente. Os animais foram submetidos à primeira aplicação do UST foi 24 horas após a cirurgia de tenotomia. A irradiação ultrassônica foi aplicada com os seguintes parâmetros: 1 MHz, modo pulsado com 20% do ciclo de trabalho (2 ms de emissão / 8 ms de intervalo), frequência de 100 Hz, 0,5 W / cm² de intensidade e ERA de 0,5 cm². A aplicação foi realizada 1x/dia. Os animais foram sacrificados após a 10ª sessão de tratamento, no 12º dia pós-operatório. Os tendões foram retirados cirurgicamente para análise da organização das fibras colágenas através do método de birrefringência (retardo óptico - OR). As fibras colágenas mostraram melhor agregação e organização no grupo UST3, UST4 e UST5 quando comparado ao GT (p<0.05) e o UST5 apresentou melhor resposta na comparação intergrupos. Conclui-se que o UST, aplicado no tempo de 5 minutos por área de radiação efetiva, apresentou a melhor dose-resposta quanto à organização das fibras colágenas no reparo tecidual de tendões de ratos...

The aim of this study was to evaluate the effect of application times 3, 4 and 5 minutes ERA of therapeutic ultrasound in the organization of the collagen fibers in rat calcaneal tendon injury. Forty male Wistar rats were used, of which 32 underwent complete tenotomy of the calcaneal tendon and were divided into 5 groups: GC without tenotomy and treatment; GT tenotomy with and without treatment; UST3, UST4 UST5 and submitted to tenotomy treated with therapeutic ultrasound at times 3, 4 and 5 minutes per effective radiating area respectively. The animals were submitted to the first application of therapy US tenotomy 24 hours after surgery. Ultrasonic irradiation was applied with the following parameters: 1 MHz, pulsed mode at 20% duty cycle (2ms transmission / 8 ms interval), frequency 100 Hz, 0.5 W / cm² intensity and ERA 0.5 cm². The application was performed 1x / day. The animals were sacrificed after the 10th treatment session, on the 12th postoperative day. The tendons were surgically removed for analysis of the organization of the collagen fibers through birefringence method (optical delay - OR). The collagen fibers showed better aggregation and organization in group UST3, UST4 and UST5 when compared to the GT (p <0.05) and UST5 showed better response in the intergroup comparison. We conclude that the UST, applied in time of 5 minutes for effective radiation area, presented the best dose-response as the organization of the collagen fibers in tissue repair of rat tendons...

Evaluation and Comparison of the Biopathology of Collagen and Inflammation in the Extracellular Matrix of Oral Epithelial Dysplasias and Inflammatory Fibrous Hyperplasia Using Picrosirius Red Stain and Polarising Microscopy: A Preliminary Study

BACKGROUND: The role of tumour inflammation and the dysplastic epithelial-stromal interactions on the nature of collagen fibres in the extracellular matrix of dysplastic epithelium is not fully understood. The present study was aimed to evaluate and compare the inflammation and pathological stromal collagen (loosely packed thin disorganized collagen) present in mild, moderate and severe epithelial dysplasias with that of inflammatory fibrous hyperplasias. The basement membrane intactness of epithelial dysplasias was also evaluated to determine if dysplastic epithelial mesenchymal interaction has any role in the integrity of stromal collagen in epithelial dysplasia. METHODS: Oral epithelial dysplasias, inflammatory fibrous hyperplasia and normal oral mucosal samples were used for the study. Packing, thickness and orientation of collagen fibres in mild, moderate and severe grades of oral epithelial dysplasias (n = 24), inflammatory fibrous hyperplasia (n = 8) and normal oral mucosal samples (n = 8) were analysed based on the polarisation of collagen fibres in picrosirius red polarising stain under polarising microscope. RESULTS: All the grades of epithelial dysplasias showed greenish yellow birefringence confirming the presence of loosely arranged pathological collagen in the presence of moderate inflammation. All the cases of inflammatory fibrous hyperplasia showed red polarisation hue and moderate inflammation. A statistically significant difference was found in the packing and orientation of collagen when epithelial dysplasias and inflammatory fibrous hyperplasia were compared (P < 0.01). When the intactness of basement membrane integrity was compared in all the groups of epithelial dysplasia, a statistically significant result was obtained (P < 0.05). CONCLUSIONS: Presence of significant amount of loosely packed thin disoriented collagen even in mild epithelial dysplasia suggests that tumourigenic factors are released to connective tissue stroma much earlier than expected. Hence we suggest considering the integrity of extracellular matrix collagen, intactness of basement membrane and inflammation associated with dysplasia along with the anaplasia of epithelial cells in the microscopic assessment of dysplastic epithelium.

Effects of M II stage oocytes zona pellucida birefringence on pregnancy outcome

OBJECTIVE@#To explore the effects of different M II stage oocytes zona pellucida birefringence on pregnancy outcome.@*METHODS@#A total of 46 couples with infertile which induced by single cause received in-vitro fertilization treatment were analyzed retrospectively, and randomly divided into the high zona birefringence (HZB)/HZB group, HZB/low zona birefringence (LZB) group and LZB/LZB group according to different oocytes zona pellucida birefringence. Intracytoplasmic sperm injection outcome was analyzed and compared.@*RESULTS@#The proportion of HZB oocytes, implantation rate and the pregnancy rate were decreased in three groups (HZB/HZB group>HZB/LZB group>LZB/LZB group) (P0.05). Logistic regression analysis showed that factors affect M II stage oocytes zona pellucida birefringence were age, basal FSH level and the LH level on the day of HCG injection. Age and FSH levels were negatively correlated with the single oocyte zona pellucida birefringence; While the LH level on the day of hCG injection was positively correlated with the single oocyte zona pellucida birefringence.@*CONCLUSIONS@#The primary influence factors on M II stage oocytes zona pellucida are age, basal FSH level and the LH level on the day of hCG injection. The birefringence value of zona pellucida can affect the pregnancy outcome.

Does women's age influence zona pellucida birefringence of metaphase II oocytes in in-vitro maturation program?

In vitro maturation [IVM] is a promising treatment option for certain infertile women. Nowadays, with the aid of PolScope, it has become possible to evaluate zona pellucida [ZP] characteristics as a parameter of oocyte quality. Moreover, quality of oocytes can be influenced by many factors, such as patient's age. The PolScope system is a non-invasive technique to assess birefringent structures such as the meiotic spindle and ZP in living oocytes. The aim was to determine the influence of the woman's age on ZP birefringence, a sign of oocyte quality, and morphology of in-vitro matured human oocytes using non-invasive polarized light [PolScope] microscopy. ZP birefringence and morphology were determined in 105 retrieved oocytes from 58 women undergoing ICSI in two age groups [>/= 30 years and <30 years]. The immature oocytes were selected and after IVM, the quality of metaphase II [MII] oocytes was assessed. The oocytes abnormalities were classified as intracytoplasmic and extracytoplasmic abnormalities. Oocyte maturation rates were significantly reduced in >/= 30 year's women [56%] in comparison with other age group [80.7%]. In addition, the ZP birefringence was significantly higher in MII oocytes in the younger group compared with the older group [76.2% vs.38.1%; p=0.00]. Following morphologic assessment, the rates of oocytes with extracytoplasmic [p=0.02] and both abnormalities [extra- and intracytoplasmic] [p=0.01] were higher in aged versus the younger women. There was a positive relationship between advanced maternal age with decreased ZP birefringence and oocyte morphological quality in in-vitro matured human oocytes

Identificación de colágeno I y III con picrosirius red/ polarización en el estroma de tumores salivales / Identification of type I and III collagen by picrosirius red/polarization of tumoral salivary stroma

El estroma juega un rol importante en los procesos tumorales de invasión y metástasis. Las fibras de colágeno tipo I son el principal componente estructural del estroma en distintos tumores. Sin embargo, hay muy pocos estudios en los tumores de glándulas salivales. Basándonos en estos antecedentes el objetivo de la presente comunicación fue estudiar las características del colágeno con Picrosirius red/polarización en tumores benignos y malignos de glándulas salivales para evaluar su posible rol en los mecanismos de progresión tumoral. Cortes histológicos de adenoma pleomórfico, carcinoma adenoide quístico y carcinoma epitelial mioepitelial se colorearon con H/E y Picrosirius red y se examinaron con microscopio de polarización. La birrefringencia del colágeno con Picrosirius/polarización resultó diferente en el estroma de los tumores malignos (carcinoma adenoide quístico y carcinoma epitelial mioepitelial), con predominio de colágeno I, en comparación con el tumor benigno (adenoma pleomórfico), con predominio de colágeno III. El diferente perfil de coloración en las fibras colágenas producidas en el estroma de los tumores analizados podría relacionarse con diferentes mecanismos de expansión tumoral, los que fueron poco estudiados en los tumores de glándulas salivales. Más estudios son necesarios para obtener resultados más concluyentes que contribuyan al diagnóstico, pronóstico y tratamiento.

The stroma plays an important rol in tumoral invasion and metastasis. Type I collagen is the main structural component of the stroma in several tumors. However, there are few studies on salivary gland tumors. Based on this background the objective of the present communication was to study collagen characteristics with picrosirius red/polarization on malignant and benign tumors of salivary glands to evaluate its posible rol in the tumoral progression mechanism. Histological sections of pleomorphic adenoma, adenoid cystic carcinoma and epithelial/myoepithelial carcinoma were stained with H/E and picrosirius red and were studied with polarization microscope. Collagen birefringence with Picrosirius/polarization was different in the malignant tumor stroma (adenoid cystic carcinoma and epithelialmyoepithelial carcinoma), with predominance of type I collagen, compared with a benign tumor (pleomorphic adenoma), with predominance of type III collagen. The different staining profile in collagen fibers produced in the benign and malignant stroma tumors analized could be related with different tumoral expansion mechanism, which were scarce studied on the salivary glands tumors. More studies are needed to obtain more conclusive results to contribute to diagnosis, prognosis and treatment.

Histopathologic Features of Triamcinolone Deposits in Refractory Steroid-Induced Glaucoma after Subtenon Triamcinolone Injection

PURPOSE: To report the removal of subtenon triamcinolone precipitates in patients with refractory steroid-induced glaucoma following subtenon triamcinolone injection. CASE SUMMARY: A 72-year-old male patient with diabetic retinopathy had cystoid macular edema in the right eye. The patient received a posterior subtenon injection of triamcinolone acetonide and developed intractable glaucoma one month after the injection in the right eye. Corticosteroid-deposit was excised three month after the injection. The intraocular pressure decreased to normal within one month after surgery and remained normal for seven months after surgery. A 42-year-old man with bilateral chronic recurrent anterior uveitis received a posterior subtenon triamcinolone acetonide injection in both eyes. The patient developed refractory steroid-induced glaucoma one month after the injection in the right eye. Corticosteroid-deposit was excised six months after the injection in the right eye. The patient's intraocular pressure decreased to normal within two weeks after surgery and remained normal. Light microscopy showed a fibrous capsule encapsulating an amorphous whitish material. The excised specimen with polarized light showed birefringence of triamcinolone crystals within an encapsulated cyst. CONCLUSIONS: Removal of subtenon triamcinolone acetonide precipitate may facilitate the management of patients developing increased intraocular pressure unresponsive to maximum tolerable medical therapy and should be considered before performing glaucoma filtration surgery.

Effect of cataract surgery on retinal nerve fiber layer thickness parameters using scanning laser polarimetry (GDxVCC).

Purpose: To study the effect of cataract extraction on the retinal nerve fiber layer (RNFL) thickness, and assessment by scanning laser polarimetry (SLP), with variable corneal compensation (GDx VCC), at the glaucoma service of a tertiary care center in North India. Materials and Methods: Thirty-two eyes of 32 subjects were enrolled in the study. The subjects underwent RNFL analysis by SLP (GDx VCC) before undergoing phacoemulsification cataract extraction with intraocular lens (IOL) implantation (Acrysof SA 60 AT) four weeks following cataract surgery. The RNFL thickness parameters evaluated both before and after surgery included temporal, superior, nasal, inferior, temporal (TSNIT) average, superior average, inferior average, and nerve fiber index (NFI). Results: The mean age of subjects was 57.6 ± 11.7 years (18 males, 14 females). Mean TSNIT average thickness (μm) pre- and post-cataract surgery was 49.2 ± 14.1 and 56.5 ± 7.6 (P = 0.001). There was a statistically significant increase in RNFL thickness parameters (TSNIT average, superior average, and inferior average) and decrease in NFI post-cataract surgery as compared to the baseline values. Mean NFI pre- and post-cataract surgery was 41.3 ± 15.3 and 21.6 ± 11.8 (P = 0.001). Conclusions: Measurement of RNFL thickness parameters by scanning laser polarimetry is significantly altered following cataract surgery. Post the cataract surgery, a new baseline needs to be established for assessing the longitudinal follow-up of a glaucoma patient. The presence of cataract may lead to an underestimation of the RNFL thickness, and this should be taken into account when analyzing progression in a glaucoma patient.

A Case of AA Amyloidosis Treated with Infliximab / 대한신장학회지

AA amyloidosis is one of the most significant complications of rheumatoid arthritis characterized by the deposition of amyloid A (AA) in multiple organs and tissues in the body. This disorder displays variable clinical symptoms depending upon the involved organ and a diagnosis is rendered through a biopsy of the affected organ, followed by staining using Congo-red which reveals an apple-green birefringence. Fundamental disease control is critical in the treatment of AA amyloidosis. Anti-tumor necrosis factor alpha (anti TNF-alpha) agents are promising in inducing clinical remission by suppressing systemic inflammation in AA amyloidosis. We report a case of AA amyloidosis in a 71 year old woman with rheumatoid arthritis that responded well to infliximab therapy.

Generalized Primary Amyloid Lymphadenopathy / 대한혈액학회지

Systemic amyloidosis is a disease that displays deposition of insoluble polymeric protein fibrils in tissues and organs. We report here on a case of a 64-year-old woman who initially presented with multiple enlarged lymph nodes. Computed tomography showed multiple enlarged lymph nodes in the mediastinal, lower cervical, supraclavicular, axillary and abdominal areas. Excision biopsy of the cervical lymph nodes and the subsequent histopathology showed amorphous eosinophilic material deposits, and these revealed apple-green birefringence on a polarizing microscopic examination on the Congo-red stained slide. The patient was diagnosed with amyloidosis and she received chemotherapy consisting of melphalan and dexamethasone. During chemotherapy, she was diagnosed with breast cancer. After modified unilateral radical mastectomy, the dexamethasone was restarted and this therapy resulted in stable disease.