Expression of Cell Cycle Markers and Proliferation Factors during Human Eye Embryogenesis and Tumorigenesis.

The expression pattern of the markers p19, Ki-67, MSX1, MSX2, PDL1, pRB, and CYCLINA2 was quantitatively and semiquantitatively analyzed in histologic sections of the developing and postnatal human eye at week 8, in retinoblastoma, and in various uveal melanomas post hoc studies by double immunofluorescence. The p19 immunoreactivity characterized retinal and/or choroidal cells in healthy and tumor tissues: expression was lower in the postnatal retina than in the developing retina and retinoblastoma, whereas it was high in epithelioid melanomas. Ki67 expression was high in the developing eye, retinoblastoma, and choroidal melanomas. MSX1 and MSX2 expression was similar in the developing eye and retinoblastoma, whereas it was absent in the postnatal eye. Their different expression was evident between epithelioid and myxoid melanomas. Similarly, PDL1 was absent in epithelioid melanomas, whereas it was highly expressed in developing and tumor tissues. Expression of pRB and CYCA2 was characteristic of developing and tumorous eye samples but not of the healthy postnatal eye. The observed expression differences of the analyzed markers correlate with the origin and stage of cell differentiation of the tissue samples. The fine balance of expression could play a role in both human eye development and ocular tumorigenesis. Therefore, understanding their relationship and interplay could open new avenues for potential therapeutic interventions and a better understanding of the mechanisms underlying the developmental plasticity of the eye and the development of neoplasms.

The Expression of Connexin 37, 40, 43, 45 and Pannexin 1 in the Early Human Retina and Choroid Development and Tumorigenesis.

The expression pattern of Connexins (Cx) 37, 40, 43, 45 and Pannexin 1 (Pnx1) was analyzed immunohistochemically, as well as semi-quantitatively and quantitatively in histological sections of developing 8th- to 12th-week human eyes and postnatal healthy eye, in retinoblastoma and different uveal melanomas. Expressions of both Cx37 and Cx43 increased during development but diminished in the postnatal period, being higher in the retina than in the choroid. Cx37 was highly expressed in the choroid of retinoblastoma, and Cx43 in epitheloid melanoma, while they were both increasingly expressed in mixoid melanoma. In contrast, mild retinal Cx40 expression during development increased to strong in postnatal period, while it was significantly higher in the choroid of mixoid melanoma. Cx45 showed significantly higher expression in the developing retina compared to other samples, while it became low postnatally and in all types of melanoma. Pnx1 was increasingly expressed in developing choroid but became lower in the postnatal eye. It was strongly expressed in epithelial and spindle melanoma, and particularly in retinoblastoma. Our results indicate importance of Cx37 and Cx40 expression in normal and pathological vascularization, and Cx43 expression in inflammatory response. Whereas Cx45 is involved in early stages of eye development, Pnx1might influence cell metabolism. Additionally, Cx43 might be a potential biomarker of tumor prognosis.

Papillary thyroid carcinoma in a hyper-functional thyroid nodule.

The authors reported the case of 69 years old woman presented with subclinical hyperthyroidism. 99m-Tc pertechnetate scan showed the abnormal focus of hot uptake in the left lobe, suggestive of a hyperfunctioning toxic thyroid nodule. Surgical treatment was advised because of the size of the nodule as a more applicable solution. Histological findings showed papillary thyroid carcinoma.

Helicobacter pylori in Otorhinolaryngology: Cause or Bystander.

The bacteria Helicobacter pylori (H. pylori) have been identified in the extragastric tissues in the head and neck. The origin and pathogenicity of these bacteria in the head and neck are not known. Gastric reflux and nasal or oral routes are the possible modes of spread. In many sinonasal, pharyngeal, laryngeal, and middle ear disorders, laryngopharyngeal reflux has been identified as a contributing or causative factor. One possible mode by which laryngopharyngeal reflux may contribute is by seeding of the extragastric mucosa with H. pylori. The clinical significance of the discovery of H. pylori in extragastric tissues in the head and neck is unclear. There is no evidence of a pathologic or active role of H. pylori in otorhinolaryngological disorders. The suggestion that the sinonasal cavities and pharynx may serve as a reservoir for H. pylori and that reinfection of the stomach occurs after eradication therapy awaits further studies for confirmation. No connection was observed between H. pylori found in the stomach and H. pylori found in the head and neck. Also, these bacteria, found in the head and neck tissues, may be accidental or innocent bystanders that do not affect the pathways of otolaryngological and gastroduodenal diseases. This review examines the evidence for a possible relationship of H. pylori with otorhinolaryngological diseases.

Immunohistochemical expression pattern of RIP5, FGFR1, FGFR2 and HIP2 in the normal human kidney development.

AIM: Present study analyses the co-localisation of RIP5 with FGFR1, FGFR2 and HIP2 in the developing kidney, as RIP5 is a major determinant of urinary tract development, downstream of FGF-signaling. METHODS: Paraffin embedded human kidney tissues of 16 conceptuses between the 6th-22th developmental week were analysed using double-immunofluorescence method with RIP5/FGFR1/FGFR2 and HIP2 markers. Quantification of positive cells were performed using Kruskal-Wallis test. RESULTS: In the 6th week of kidney development RIP5 (89.6%) and HIP2 (39.6%) are strongly expressed in the metanephric mesenchyme. FGFR1 shows moderate/strong expression in the developing nephrons (87.3%) and collecting ducts (70.5%) (p < 0.05). RIP5/FGFR1 co-localized at the marginal zone and the ureteric bud with predominant FGFR1 expression. FGFR2 (26.1%) shows similar expression pattern as FGFR1 (70.5%) in the same kidney structures. RIP5/FGFR2 co-localized at the marginal zone and the collecting ducts (predominant expression of FGFR2). HIP2 is strongly expressed in collecting ducts (96.7%), and co-localized with RIP5. In 10th week, RIP5 expression decrease (74.2%), while the pattern of expression of RIP5 and FGFR1 in collecting ducts (33.4% and 91.9%) and developing nephrons (21.9% and 32.4%) (p < 0.05) is similar to that in the 6th developmental week. Ureter is moderately expressing RIP5 while FGFR1 is strongly expressed in the ureteric wall. FGFR2 is strongly expressed in the collecting ducts (84.3%) and ureter. HIP2 have 81.1% positive cells in the collecting duct. RIP5/FGFR1 co-localize in collecting ducts and Henley's loop. CONCLUSIONS: The expression pattern of RIP5, FGFR1, FGFR2 and HIP2 in the human kidney development might indicate their important roles in metanephric development and ureteric muscle layer differentiation through FGF signaling pathways.

Expression and localization of DAB1 and Reelin during normal human kidney development.

AIM: To explore the spatial and temporal expression patterns of DAB1 and Reelin in the developing and postnatal healthy human kidneys as potential determinants of kidney development. METHODS: Paraffin-embedded fetal kidney tissue between the 13/14th and 38th developmental weeks (dw) and postnatal tissue at 1.5 and 7 years were stained with DAB1 and Reelin antibodies by double immunofluorescence. RESULTS: During the fetal kidney development and postnatal period, DAB1 and Reelin showed specific spatial expression pattern and diverse fluorescence intensity. During the fetal period, DAB1 was strongly expressed in the distal convoluted tubules (DCT), with strong reactivity, and diversely in the proximal convoluted tubules (PCT) and glomeruli. In the postnatal period, DAB1 expression decreased. The strongest Reelin expression in early fetal stages was observed in the PCT. In the postnatal period, Reelin expression decreased dramatically in all observed structures. These two markers were colocalized during early developmental stages, mostly in PCT, DCT, and podocytes. CONCLUSION: The appearance of DAB1 and Reelin during fetal kidney development confirms their potential significant role in the formation of kidney structure or function. High DAB1 expression in the DCT implies its regulatory role in tubular formation or function maintenance during development. Reelin was highly expressed in human kidneys at early fetal stages, mostly in the PCT, while at later fetal stages and postnatal period its expression decreased.

Immunohystochemical Expression of Vitamin D Receptor in Development Stages of Colorectal Carcinoma.

BACKGROUND: Chemo preventive and antitumor role of vitamin D is manifested through genetic and non genetic ways with a powerful antproliferatory and proapopoptic effect, which is proven by numerous epidemiologic studies. The genetic activity of vitamin D is determined through vitamin D receptors (VDR), a member of stero-thyreoidal family of nuclear receptors, which with vitamin D form a cell nucleus complex responsible for the chemo preventive and antitumor effect. VDR in tissue cells is present in the cytoplasm and the nucleus and manifests its genetic activity after transfer from the cytoplasm to the nucleus. The mechanisms for the transport and genetic control of the transport of VDR from cytoplasm to the nucleus in not yet completely understood. SUBJECTS AND METHODS: By using immunohystochemistry we are evaluating the correlation of cytoplasmic and nuclear expression of VDR during different stages of colorectal carcinoma: normal colorectal mucosa, hyperplasic polyp, low grade adenoma (LGD), high grade adenoma (HGD) and colorectal cancer. RESULTS: Our results confirm that the nuclear VDR expression is strongest in normal colorectal mucosa and in hyper plastic polyps, is gradually weakened in low and high grade adenoma while it is extremely weak or absent in colorectal carcinoma. At the same time the expression of cytoplasm VDR is weakest in normal colorectal mucosa and hyper plastic polyps while it grows during the adenoma stage and is most expressed during colorectal carcinoma. CONCLUSION: We conclude that vitamin D has a strong chemo preventive and antitumor effect in normal colorectal mucosa and hyper plastic polyps, while its antitumor and chemopreventive effect is progressively weakened and ultimately absent in colorectal carcinoma.

The influence of dreinage in the treatment of chronic pilonidal sinus disease using midline closure.

Unsatisfactory postsurgical end results in the treatment of chronic pilonidal sinus disease caused by long term healing, pain, inconvenience and recurrences of the lesion still remain problem after various surgical techniques have been described and used. This study was designed to show results of several aspects of midline closure technique with respect for surgical complications, hospitalization length and recurrence. This study included 90 patients with pilonidal sinus treated by surgery with midline closure at the department of General surgery of Clinical Hospital Mostar between January 2004 and January 2009. The patients were divided in three groups considering the type of drainage used. Data collected retrospectively included demographics, positive diagnosis of chronic pilonidal sinus, operative technique, type of drainage, complications, hospital stay and recurrence of the disease. Mean hospital stay after excision and midline closure technique was 4.68 days Complication rate was 18/90 (20%). Infection occurred in 18 patients (20%), dehiscence occurred in 10 patients (11.1%). Mean recurrence rate during follow up period was 12/90 (13.3%); range, 24-84 months. There was no statistically significant difference among three groups in hospital stay length (p>0.05), in complication rate (χ(2)=1.66, p>0.05), nor in recurrence rate (χ(2)=1.91, p>0.05). Statistically significant difference was shown between complication rate among non drained and actively drained patients (χ(2)=1.11, p<0.05). Primary midline closure is not a satisfactory method in the treatment of chronic pilonidal sinus, especially without drainage, leading to numerous complications and high rate of recurrence.

The Epidemiological Characteristics of Hepatitis B in Croatia: The Results of the Prevention.

This study shows epidemiological characteristics and preventive measures implemented for the prevention and control of hepatitis B infections in Croatia. We analyzed the data from obligatory infectious disease reports and notifications of death due to infectious diseases, data on the hepatitis B infections in Croatia, and data collected by survey of the population. The average prevalence of the disease is 3.67 per 100,000 annually. All age groups are affected, but still a higher rate of the disease is found in the age groups from 15-19 and 20-29 years of age. Hepatitis B disease is 1.4 times more likely in men than in women. For the past 18 years, the average rate of mortality was 0.2%. The incidence of HbsAg-positive donors of blood is within the range of 0.65% in 1992 to 0.012% in 2011. The largest part of preventive measures implemented in Croatia against hepatitis B is predicted and required by legislation. The registrations of acute and chronic carriers of the virus are obligatory. High-risk groups have started being vaccinated since 1992. The obligatory vaccination of infants was introduced in the mandatory vaccination program in 2007. Routine testing of blood exclusively from voluntary donors for HbsAg presence is obligatory. The non-governmental organization "Help" created for intravenous drug users, along with the "Harm reduction" program implemented hepatitis B, C, and HIV/AIDS prevention program in 1995. In order to gain a better understanding of epidemiological characteristics of hepatitis B in Croatia, the specifics of its dynamics in small communities are required since the research of Croatian public health officials and researchers have shown that hepatitis B is spread in different ways.

Expression of Ki-67, Oct-4, γ-tubulin and α-tubulin in human tooth development.

AIMS: To analyze factors controlling cell proliferation and differentiation, and appearance of primary cilia during the cap and bell stages of incisor or/and canine human enamel organs. MATERIALS AND METHODS: Qualitative and quantitative analysis of proliferating Ki-67 positive cells and expression of γ-tubulin, α-tubulin and Oct-4 was immunohistochemically analyzed in the cap an bell stages of 10 developing human incisor and canine germs, 8-21 weeks old. RESULTS: During the analyzed period, ratio of Ki-67 positive cells changed in outer enamel epithelium from 48.86% to 24.52%, in inner enamel epithelium increased from 56.11% to 60.06% and then dropped to 44.24%. While in dental papilla proliferation first increased from 46.26% to 55.45%, and then dropped to 22.08%, a constant decrease of proliferation characterized enamel reticulum (from 46.26% to 15.49%). Strong cytoplasmic Oct-4 expression characterized epithelial parts of enamel organ, particularly the differentiating ameloblasts. During further development, Oct-4 expression shifted to both nuclear and cytoplasmic expression in mesenchymal tooth components. Primary cilia characterized most of the cells in developing enamel organ. While non-ciliated (proliferating) cells mainly contained two centrioles (γ-tubulin), the primary cilia (α-tubulin) were arising from basal bodies (γ-tubulin) of non-proliferating cells. CONCLUSIONS: We suggest that increase in cell proliferation enables growth of enamel organ, while its selective decrease leads to disintegration of some tooth parts. Drop of proliferation coincided with initiation of ameloblast and odontoblast differentiation. Additionally, cell differentiation was accompanied by increased expression of Oct-4 and probably by signalling via primary cilia, both regulating processes of cell proliferation and differentiation.

Polypoid angiomyofibroblastoma tumor of nasal cavity: case report.

We report an extremely rare case and localization of polypoid angiomiofibroblastoma tumour, a case report and review of the world literature concerning angiomiofibroblastoma tumour. We present the case of a 74-year-old man who underwent left anterior and posterior ethmoidectomy with extirpation of tumour mass from left nasal cavity, epipharinx and left sphenoid sinus. The prognosis for this group of tumour is good and patient didn't receive any kind of therapy except surgical treatment. To our knowledge, this is a rare report in the world literature of polypoid angiomiofibroblastoma tumour of nasal cavity. This case indicates that angiomiofibroblastoma tumour of nasal cavity and paranasal sinuses is a rare disease including its localisation which otolaryngologists should be aware of and one which should be included in the differential diagnosis of tumours involving sinonasal tract.

Immunolocalization of nestin, mesothelin and epithelial membrane antigen (EMA) in developing and adult serous membranes and mesotheliomas.

The spatial and temporal distribution of epithelial membrane antigen (EMA), mesothelin and nestin was immunohistochemically analyzed in developing and adult human serous membranes and mesotheliomas in order to detect possible differences in the course of mesenchymal to epithelial transformation, which is associated with differentiation of mesothelial cells during normal development and tumorigenesis. Pleura and pericardium developing from the visceral mesoderm gradually transform into mesothelial cells and connective tissue. EMA appeared in mesothelium of both serous membranes during the early fetal period, whereas during further development, EMA expression was retained only in the pericardial mesothelium. It increased in both pleural mesothelium and connective tissue. Mesothelin appeared first in pericardial submesothelial cells and later in surface mesothelium, while in pleura it was immediately localized in mesothelium. In adult serous membranes, EMA and mesothelin were predominantly expressed in mesothelium. Nestin never appeared in mesothelium, but in connective tissues and myocardial cells and subsequently decreased during development, apart from in the walls of blood vessels. Mesothelial cells in the two serous membranes developed in two separate developmental pathways. We speculate that submesothelial pericardial and mesothelial pleural cells might belong to a population of stem cells. In epithelioid mesotheliomas, 13% of cells expressed nestin, 39% EMA and 7% mesothelin.

Expression of nestin, mesothelin and epithelial membrane antigen (EMA) in developing and adult human meninges and meningiomas.

The spatial and temporal pattern of appearance of nestin, epithelial membrane antigen (EMA) and mesothelin proteins was immunohistochemically determined in the cells of normal developing and adult human meninges and meningiomas. Human meninges developed as two mesenchymal condensations in the head region. The simple squamous epithelium on the surface of leptomeninges developed during mesenchymal to epithelial transformation. Nestin appeared for the first time in week 7, EMA in week 8, while mesothelin appeared in week 22 of development. In the late fetal period and after birth, nestin expression decreased, whereas expression of EMA and mesothelin increased. EMA appeared in all surface epithelial cells and nodules, while mesothelin was found only in some of them. In adult meninges, all three proteins were predominantly localized in the surface epithelium and meningeal nodules. In meningothelial meningiomas (WHO grade I), EMA was detected in all tumor cells except in the endothelial cells, mesothelin characterized nests of tumor cells, while nestin was found predominantly in the walls of blood vessels. The distribution pattern of those proteins in normal meningeal and tumor cells indicates that nestin might characterize immature cells, while EMA and mesothelin appeared in maturing epithelial cells. Neoplastic transformation of these specific cell lineages contributes to the cell population in meningiomas.

Expression of estrogen and progesterone receptors in human ductal invasive breast carcinoma not otherwise specified: is there any difference between premenopausal and postmenopausal women?

Determination of hormone receptors is of utmost importance in planning therapy in patients with breast cancer. The aim of the study was to assess the expression of estrogen (ER) and progesterone (PR) receptors in ductal invasive breast carcinoma not otherwise specified (NOS) according to patient menopausal state and tumor histopathology. The study included 549 patients treated at University Department of Surgery, Rijeka University Hospital Center, between January 1, 2000 andJanuary 1, 2005. The patients were diagnosed with breast cancer and underwent mastectomy. ER and PR status was determined by immunohistochemistry. Study results showed no statistically significant differences in the expression of ER and PR, tumor size and grade of histologic differentiation between premenopausal and postmenopausal women. However, tumor size and grade of histologic differentiation differed significantly according to the expression of hormone receptors. Tumors greater than 5 cm in size were mostly ER- in premenopausal (P = 0.012) and PR- in postmenopausal (P = 0.044) patients. Poorly differentiated cancers were associated with ER-PR-status in both premenopausal and postmenopausal patients (P < 0.001). Hormone dependent tumors (ER+PR+) were of smaller diameter and lower histologic grade, while hormone independent tumors (ER-PR-) had greater diameter and higher histologic grade, the difference being statistically significant (P = 0.004 and P < 0.001, respectively). Study results on the characteristics of ductal invasive carcinoma according to hormone status were consistent with those described in the literature. Considering controversies about the role of steroid receptors in endocrine therapy response, our future objective is assessment of the 5-year prognosis in our patients.

Apoptotic and anti-apoptotic factors in early human mandible development.

The regulators of apoptosis Bcl-2, Bax, caspase-3, p53, and Hsp70 were analyzed immunohistochemically in the developing human mandible of eight human conceptuses from weeks 5 to 10 of gestation. During this period, all proteins displayed an increased pattern of expression in the mandible ectomesenchyme and in newly formed bone, except for caspase-3, which showed decreased expression in the ectomesenchyme, but appeared first in the ossification zone at the 7th wk of development. Simultaneously, the oral epithelium showed weak (p53) to strong (hsp70) expression of all proteins investigated, while in Meckel's cartilage cells, bcl-2 was expressed weakly and hsp70 was expressed moderately. Cells on the surface of the forming bone were predominantly bax positive, and only occasionally bcl-2 positive. Only a few cells on the surface and inside the bony spicules co-expressed bax and bcl-2. Terminal deoxynucleotidyl transferase (TdT)-mediated biotin-dUTP nick-end labelling (TUNEL)-positive cells were found to be apoptotic osteoblasts. The expression of all proteins investigated changed dynamically during early mandible development and the subsequent differentiation of Meckel's cartilage and bone. While interactions between those factors might be associated with the survival of Meckel's cartilage, in the ossification zone they might participate in the control of cell numbers, mineralization, and bone remodelling. Among many other factors, precise orchestration of pro- and anti-apoptotic factors contributes to normal mandible development.

Q fever in Croatia: war-induced changes in epidemiological characteristics.

The article describes the epidemiological characteristics of Q fever in Croatia, during the period before and after the Homeland War. In the ten years prior to the Homeland War (1983-1992) 1053 cases of Q fever were recorded, 16.2% (171) of which on islands and in coastal areas. In the period after the Homeland War (1995-2008), a total of 654 cases was recorded, 59.9% (392) of which on islands and in coastal areas. In addition to reduced incidence, geographic distribution of the disease also changed. Before the war, the highest morbidity rate was recorded in the Sisak-Moslavina County. After the war, the Split-Dalmatia County recorded the highest morbidity rate because in the post-war period sheep from Bosnia and Herzegovina went to this county for winter grazing. The disease might be relevant to the Croatian Army and other armed forces that stay in Croatia as part of NATO forces.