First trimester maternal serum biochemical markers of aneuploidy in pregnancies with abnormally invasive placentation.

OBJECTIVE: To examine whether differences exist in routine first trimester maternal serum screening analyte measurements between normal pregnancies, placenta praevia and abnormally invasive placentation (AIP). DESIGN: Multidisciplinary audit. SETTING: Associated university teaching hospital with 9000 annual deliveries. POPULATION: Five hundred and sixteen pregnancies in total, including 344 normal controls, 17 with AIP and 155 placenta praevia cases. METHODS: Comparison of maternal serum free ßhCG and PAPP-A MoMs distribution in pregnancies with abnormally invasive placentation, placenta praevia and normal controls, after correcting for known confounding factors between October 2005 and September 2013. Data from a previously published first trimester AIP and biochemistry study were combined with our study data and compared in the above way to complete the analysis. MAIN OUTCOME MEASURES: Differences in first trimester maternal serum PAPP-A and free ßhCG in AIP, placenta praevia, and normal pregnancies. RESULTS: Median free ßhCG MoM in the control group was 1.04, and 1.08 (P = 0.859) in the placenta praevia group compared with 0.81 in the AIP group (P = 0.06). Median PAPP-A MoM was 1.01 in the control group and 1.05 (P = 0.83) in praevia, compared with 1.22 in AIP cases (0.16). The combined AIP dataset gave an overall PAPP-A median MoM of 1.40, and free ßhCG of 0.85. Both markers showed a significantly different distribution from controls (PAPP-A P = 0.002 and free ßhCG P = 0.031). CONCLUSIONS: There may be differences between first trimester maternal serum biochemical markers between normal pregnancies and those complicated by abnormally invasive placentation. If upheld, this may provide useful information for the early identification of abnormally invasive placentation. More studies are required.

Squamous cell carcinoma growth in mice and in culture is regulated by c-Jun and its control of matrix metalloproteinase-2 and -9 expression.

Squamous cell carcinoma (SCC) is an invasive malignancy of epidermal keratinocytes. Surgical excision is currently the main treatment; however, this can cause scarring and disfigurement. There is accordingly, an acute need for alternative strategies to treat SCC. The transcription factor c-Jun is expressed in human SCC and another common form of invasive skin cancer, basal cell carcinoma together with the mitogenic marker-proliferating cell nuclear antigen. Here, we have employed DNAzymes (catalytic DNA molecules) targeting c-Jun (Dz13) to inhibit c-Jun expression in SCC cells. Dz13 inhibits SCC proliferation and suppresses solid SCC tumor growth and tumor angiogenesis in severe combined immunodeficient mice. We further demonstrate that Dz13 inhibits c-Jun, together with matrix metalloproteinase (MMP)-2 and MMP-9 expression in the tumors, consistent with DNAzyme inhibition of MMP-2 and MMP-9 gelatinolytic activity by zymography. Dz13 also suppressed the expression of vascular endothelial growth factor and fibroblast growth factor-2 in the tumors. These findings demonstrate that c-Jun regulates SCC growth and suggest that DNAzymes targeting this transcription factor may potentially be useful as inhibitors of cutaneous carcinoma.

Histologic assessment of equivocal cytologic smears.

OBJECTIVE: To determine if the conversion of equivocal cytologic smears into histologic sections would provide additional diagnostic information in those cases in which it was difficult to obtain additional cytologic samples. STUDY DESIGN: Over a year, eight equivocal cytologic smears were converted to histologic sections by removing the coverslips, rehydrating the smears, scraping off the smears into centrifuge tubes, making cell blocks, and sectioning and staining the blocks. RESULTS: The histologic sections enabled cell patterns to be studied and special staining (including immunoperoxidase studies) to be performed. In the eight cases studied, the additional information provided by histology led to a definitive diagnosis in six (75%). CONCLUSION: When cytologic findings are equivocal and it is difficult to obtain additional cytologic samples, conversion of the smears into histologic sections may provide additional information for diagnosis.

Problems in the histological assessment of hydatidiform moles: a study on consensus diagnosis and ploidy status by fluorescent in situ hybridisation.

Hydropic villi in products of conception continue to pose a diagnostic problem for the anatomical pathologist. It is important to distinguish between complete hydatidiform mole (CM), partial hydatidiform mole (PM) and hydropic degeneration (HD), as hydatidiform moles (especially CM) have a tendency to develop persistent trophoblastic disease. Several studies have demonstrated interobserver variability in the diagnosis of the three conditions, but there have been no studies testing the accuracy of the consensus diagnosis of pathologists experienced in the field. In this study four anatomical pathologists with experience in diagnosing hydatidiform moles selected five cases of HD, seven cases of PM and ten cases of CM on the basis of consensus diagnosis using established criteria. Ploidy studies were done on these 22 cases using fluorescent in situ hybridisation. The 15 cases of HD and CM were diploid, confirming the histological diagnosis. However only five of the seven cases of PM were triploid, the other two being diploid. Review of these two diploid cases showed a mixture of small and large villi with moderate to marked trophoblastic proliferation. On the basis of the significant trophoblastic proliferation and the DNA information, the two cases were reclassified as early complete moles. This study demonstrates that even pathologists experienced in the field have difficulty separating PM from CM. The findings suggest that, in the absence of DNA information, a lesion with hydropic villi showing moderate to marked trophoblastic proliferation should be classified as a complete mole, even if there is a mixture of small and large villi. Ploidy studies are an important adjunct to histological diagnosis, especially when there is an overlap of features.

Combined cytological and histological diagnosis of hepatocellular carcinoma in ultrasonically guided fine needle biopsy specimens.

This study evaluates the usefulness of a combined cytological and histological approach to the diagnosis of hepatocellular carcinoma (HCC) when applied to fine needle biopsy specimens obtained under ultrasonic guidance. The material, aspirated from 51 focal liver lesions, was handled in such a way that there was sufficient material for both cytological and histological (cell block) assessment. Of the 29 cases of HCC studied, a confident cytological diagnosis was made in 23 (79%). In the remaining six cases, the cytological features were considered to be suspicious but not diagnostic of HCC. Examination of cell blocks in the six cases enabled a confident diagnosis of HCC to be made in all cases. This was due to the supplementary visual information provided by the histological features, particularly the pattern of arrangement of the tumour cells.

Concomitant mucinous tumors of appendix and ovary. Result of a neoplastic field change?

BACKGROUND: Mucinous tumors of the appendix and ovary are known to occur together in association with pseudomyxoma peritonei. It has been postulated that this association may be attributable to the development of independent tumors or to metastasis from one site to another. METHODS AND RESULTS: This article reports two patients with concomitant mucinous ovarian and appendiceal tumors in the absence of pseudomyxoma peritonei. CONCLUSIONS: The evidence suggests that these tumors are independent primary neoplasms that develop as a result of neoplastic field change that affects colonic-type epithelium.

An assessment of the value of frozen sections in gynecological surgery.

In 203 consecutive gynecological operations where frozen sections were performed, 35.6% were from conditions of the ovary, 22.7% from the cervix, 18.2% from the endometrium, and 11.4% from the vulva. There were 0.5% false-positive, 1.0% false-negative and 2.0% deferred diagnosis. Incorrect interpretation was the cause of the single false-positive diagnosis, while the false-negative diagnoses were due to errors in block selection. The deferred diagnoses mainly occurred in gynecological conditions where diagnosis was difficult, required extensive sampling or a formal mitotic count. As in other surgical fields, gynecological frozen sections were used principally to guide the extent of surgery. The most valuable frozen sections were in those instances where the operation was affected most. These were on lymph nodes in cases of carcinoma of the vulva and cervix, myometrial lesions in young women where myomectomy was being considered, and ovarian tumours to distinguish primary from secondary tumours. Occasionally, frozen sections were also found useful to establish margins of vulval and cervical tumours, in hysterectomy specimens of endometrial carcinomas to determine prognostic factors, and in suspected recurrences and metastases of tumours to determine the adequacy of the biopsy material. Frozen sections in obviously benign conditions, e.g., ovarian cysts without papillary or solid areas, were found to be unnecessary. Frozen sections are contraindicated when only a small amount of crucial material is available, as the paraffin diagnosis may be compromised. Pathologists should have a clear idea of the role of frozen sections in gynecological surgery and work closely with the surgeon in the management of gynecological oncology patients.

Morphologic differences in the pattern of liver infiltration between systemic AL and AA amyloidosis.

Biopsy and necropsy tissue from 31 unselected patients with systemic amyloidosis, in which there was histologic evidence of liver involvement, were reviewed with reference to the location and pattern of amyloid deposition in the liver. Amyloidosis was classified into AA and AL types on the basis of immunohistochemistry and permanganate reaction of the amyloid deposits. Nineteen were categorized as AA (secondary) and 12 as AL (primary) amyloidosis. Deposition of AA amyloid was limited to the walls of vessels in the portal tract, constituting a "vascular" pattern. In AL amyloidosis, the deposits exhibited a "sinusoidal" pattern in that they were seen along hepatic sinusoids as well as in vessel walls. This difference was statistically significant (P less than .001). The histologic pattern of liver infiltration offers a valuable clue in the classification of systemic amyloidosis and provides information that may be useful in the selection of patients for therapy.

The prognostic significance of grading in borderline mucinous tumors of the ovary.

Fifty-three stage I borderline mucinous tumors of the ovary were placed into four histologic grades according to a simple system of grading based on degree of cell layering, nuclear characteristics, and mitotic count. Three patients died of recurrence and spread of their tumors. All three patients had grade 4 tumors, suggesting that there may be some prognostic value in grading borderline mucinous ovarian tumors.

Sarcomatous change in granulosa cell tumor.

Granulosa cell tumors have a tendency for indolent growth and late recurrence. The present case of granulosa cell tumor in a 67-year-old woman is the first, to the authors' knowledge, to be reported as showing sarcomatous transformation with a rapid fatal course. Both the granulosa cell and fibrothecomatous elements appear to have undergone sarcomatous change with some evidence to suggest rhabdomyosarcomatous differentiation. The sarcomatous component was present in metastatic deposits.

Race-related morphologic variations in hepatocellular carcinoma.

In West Malaysia, hepatocellular carcinoma (HCC) is common in the Chinese and in the members of the Senoi aboriginal tribe, two racial groups with diametrically opposite life-styles. Certain fundamental differences exist between the liver tumors in the two races. In the Senoi, the tumor occurs in a younger age group and there is a greater male preponderance than in the Chinese. There is also a very close relationship between hepatitis B virus infection, chronic active hepatitis, cirrhosis, liver cell dysplasia, and HCC in the Senoi and the tumors generally present as multiple nodules studding both lobes of the liver. In the Chinese, although a relationship between hepatitis B virus infection, HCC, and cirrhosis exists, this association is not as strong as in the Senoi and the tumors are generally large and solitary. The data suggest that, although the hepatitis B virus is probably an important oncogenic agent in both racial groups, there may be a difference in the pathogenesis of HCC in the two races.

Copper-binding protein in liver cells.

The patterns and incidences of orcein-positive granules of copper-binding protein (CBP) in 2,531 liver biopsy specimens from children and adults with a large variety of liver diseases are reported. Fetal and neonatal livers have high physiologic levels of copper and CBP, which fall to within the adult range by the third to the sixth month of life. Therefore, in liver specimens from children less than 6 months of age, it was not possible to determine whether the orcein-positive granules present represented physiologic or pathologic deposits of CBP. In adults and in children older than 6 months of age, CBP granules were found almost exclusively in association with four main groups of liver diseases: Wilson's disease, chronic biliary disorders, cirrhosis/extensive fibrosis, and primary liver tumors. Orcein-positive granules were never found in patients with acute liver disease. The granules were extremely helpful in distinguishing chronic biliary diseases from acute cholestatic and hepatic disorders, primary biliary cirrhosis from chronic active hepatitis, and primary liver tumors from metastatic tumor deposits.

In vitro diagnosis of cow's milk protein sensitive enteropathy by organ culture method.

The criteria that are used at present to diagnose cow's milk protein sensitive enteropathy (CMPSE) are based on an in vivo milk challenge which can be hazardous and life threatening. We have used an organ culture model to determine the usefulness of this technique in establishing the diagnosis of CMPSE on the basis of a single biopsy with in vitro milk challenge. Fourteen infants with diarrhoea clinically suspected to have CMPSE were studied prospectively. On the basis of milk challenge studies seven infants had CMPSE. They had clinical reaction to cow's milk with associated histological changes and depression of alkaline phosphatase levels in the jejunal mucosa. In all seven cases parallel changes in alkaline phosphatase levels were noted in the organ culture specimens of initial biopsy subjected to in vitro challenge. The seven control infants tolerated cow's milk and did not have histological changes. The alkaline phosphatase levels were moderately increased in the jejunal mucosa in five of the seven infants. The alkaline phosphatase levels in the organ culture specimens of initial biopsy were increased after in vitro challenge in all seven infants. This study suggests that organ culture methods may be useful in the vitro diagnosis of CMPSE, and also obviate the need for in vivo oral milk challenges and repeated biopsies.

An appraisal of the relationship between primary hepatocellular carcinoma and hepatitis B virus.

The association between hepatitis B virus infection and primary hepatocellular carcinoma is reviewed. On the basis of serological and tissue examination there is a close link between virus infection and the tumour. While there is evidence to favour an oncogenic role for virus this is not conclusive, and other possible explanations for the relationship are discussed.

Is jejunal biopsy really necessary in cow's milk protein intolerance?

Thirty-nine infants suspected of having cow's milk protein intolerance (C.M.P.I.) were investigated, and jejunal biopsies were performed before and after challenge with cow's milk. Thirty patients had significant jejunal mucosal damage after milk challenge, but symptoms of diarrhoea and vomiting developed in only twenty-two. The patients with symptoms were subsequently managed on a diet free from cow's milk until tolerance developed. However, the eight infants without symptoms (but with jejunal mucosal damage) made satisfactory clinical progress, with adequate weight-gain, on a diet of cow's milk. Repeat jejunal biopsy specimens from two of these patients showed that there had been a definite improvement since the immediate post-challenge biopsy specimens were taken. Most patients with C.M.P.I. who need to be treated with a diet from which cow's milk has been eliminated may be detected by clinical means alone, and the remainder may continue on a cow's milk diet unless or until symptoms develop. There seems to be no clinical justification for routine jejunal biopsy in infants in whom C.M.P.I. is suspected.

HBAg-positive chronic liver disease associated with cirrhosis and hepatocellular carcinoma in the Senoi.

Necropsy and clinical data show that primary hepatocellular carcinoma (PHC) is the commonest cancer among the Senoi (a Malaysian aboringine group). The other aboringine tribes do not appear to have this high predilection for liver cancer. In the necropsy series, PHS was present in 10 out of 22 Senoi patients with cirrhosis. All the 22 livers contained hepatocytes that stained with Shikata's orcein stain and specific immunoperoxidase and immunofluorescent stains for hepatitis B antigen (HBAg). This observation raises the strong possibility that hepatitis B may be an important etiologic factor in the development of cirrhosis and PHC in the Senoi. The reason for the high susceptibility of the Senoi for HB virus infection is not clear, and the role of aflatoxin in the pathogenesis of PHC in the Senoi has yet to be determined. That the Senoi are a numerically small community, maintaining their own unique dietary and social customs and living in readily accessible areas in the Malaysian jungle, makes them an ideal population for the study of factors in the etiology of liver cancer.

Methods for detection of hepatitis B surface antigen in paraffin sections of liver: a guideline for their use.

Methods for the localisation of hepatitis B surface antigen (HBsAg) in paraffin sections of the liver include the detection of ground-glass hepatocytes and the use of Shikata's orcein stain, and of immunoperoxidase and immunofluorescent techniques. A comparative study of the different methods on 20 livers shows the orcein stain to be the method of choice for routine use. The Shikata stain is not only specific but is relatively inexpensive, easily performed, and stains out distinct cytoplasmic inclusions even in stored formalin-fixed livers, old paraffin blocks, and autolysed livers. Since HBsAg is irregularly distributed in the liver, adequate sampling is necessary to prevent false negative; when sufficient tissue is available at least five blocks should be examined before a case is labelled as HBsAg-negative.