Squamous cell carcinoma: pain as a clue to increased tumour diameter, increased invasion depth, the grade of differentiation, acantholysis and perineural invasion.

BACKGROUND: Pain may be associated with actinic keratosis (AK), intraepidermal carcinoma (IEC) and invasive squamous cell carcinoma (SCC), which may all display high-risk features. AIM: To examine variation in pain frequency associated with these three conditions, and assess their invasive SCC surface diameter, invasion depth, grade of differentiation, presence of acantholysis and perineural invasion (PNI). METHODS: Pain was prospectively recorded for consecutive cases of AK, IEC and SCC from three institutions in Australia during the period 2016-2018. RESULTS: Pain with palpation was recorded with 15.8% of AK (n = 30/190), 15.1% of IEC (n = 345/299) and 29.0% invasive SCC (n = 247/853). Pain without palpation was respectively 1.1% (2/190), 4.0% (12/299) and 6.7% (57/853). Invasive SCC with increased surface diameters and deeper invasion recorded increased pain frequency. Pain did not vary significantly by the grade of differentiation in males. In females, well-differentiated SCC recorded more pain (45.4%; n = 473) than poorly differentiated SCC (9.1%; n = 11). Acantholytic SCC recorded more pain 48.7% (n = 29) than nonacantholytic SCC 35.2% (n = 824). Three out of five cases of PNI recorded pain. Pain intensity was not recorded, which was a limitation. CONCLUSION: Pain presence increases from AK to invasive SCC. Pain was more frequent in invasive SCC with increased surface diameter, deeper invasion, acantholysis and PNI. Pain frequency did not vary between the grades of differentiation in males. In females, pain was less frequent in poorly differentiated than in well-differentiated SCC.

Invasive squamous cell carcinoma: comparison of differentiation grade and tumour depth by anatomical site in 1666 tumours.

BACKGROUND: Invasive squamous cell carcinomas (SCCs) presents with different grades of differentiation and depths of invasion. AIM: To compare the grade of differentiation, tumour diameter and tumour depth by anatomical site in invasive SCC. METHODS: Retrospective clinical and histopathological data on consecutive cases of SCC came from a clinic in Sydney, Australia were assessed. A multinomial logistic regression model was applied to compare grades of differentiation by age, sex, anatomical sites, and histological tumour maximum diameter and depth. RESULTS: In total, 1666 SCCs were identified, including 82.1% (n = 1367) well-differentiated, 13.3% (n = 222), moderately differentiated and 4.6% (n = 77) poorly differentiated SCCs. Patients with poorly differentiated tumours were more likely to be older and male (both P < 0.001). The most common site for poor differentiation was the scalp in men (n = 12; 15.6%) and the cheek or chin in women (n = 7; 9.1%). In the multivariate model, compared with well-differentiated SCC, older age was significantly associated with poorly and moderately differentiated SCC (P < 0.01 and P = 0.02, respectively). Larger tumour diameters were related to poor differentiation (P = 0.03). Ear, forehead and chest sites had increased tumour depth and poor differentiation. CONCLUSIONS: This study found increased rates of poorly differentiated SCC on the forehead and cheek for both sexes, while men displayed increased rates of poorly differentiated SCC on the bald scalp and the ears. Tumour diameter and depth increased as tumours varied from well-differentiated to moderately differentiated and from moderately differentiated to poorly differentiated. An increase in depth and increased prevalence of poorly differentiated tumours were found on the ears for men and on various facial sites for both sexes.

Acantholytic invasive squamous cell carcinoma: tumor diameter, invasion depth, grade of differentiation, surgical margins, perineural invasion, recurrence and death rate.

BACKGROUND: Squamous cell carcinoma (SCC) may present with or without the feature of acantholysis. METHODS: Investigate invasive acantholytic SCC by microscopic maximum tumor surface diameter, depth of invasion, grade of differentiation, perineural invasion (PNI) and percentage of acantholysis. Assess recurrence following excision. RESULTS: A total of 1658 consecutive invasive SCC cases were examined, comprising 4.9% acantholytic SCC. Median tumor microscopic maximum diameter was 8 mm for acantholytic SCC and 7.3 mm for non-acantholytic SCC. Median tumor invasion depth was 1.0 mm for acantholytic SCC and 1.5 mm for non-acantholytic SCC. Well, moderate and poor differentiation were not significantly different between acantholytic SCC and non-acantholytic SCC. One PNI case was found in 82 acantholytic SCC cases. A total of 77 acantholytic SCC cases were followed up over a median 25 months finding histologic proven recurrence at three acantholytic SCC excision sites. CONCLUSIONS: Acantholytic SCC were more likely to be located on head sites with less median depth than non-acantholytic SCC. Increasing percentage of acantholysis within acantholytic SCC was not associated with a shift towards poor differentiation. Histologic margins of 1.2 mm may adequately excise small acantholytic SCC. No recorded deaths, low PNI and low recurrence rates suggests acantholytic SCC is low-risk.

Pine oil effects on chemical and thermal injury in mice and cultured mouse dorsal root ganglion neurons.

A commercial resin-based pine oil (PO) derived from Pinus palustris and Pinus elliottii was the major focus of this investigation. Extracts of pine resins, needles, and bark are folk medicines commonly used to treat skin ailments, including burns. The American Burn Association estimates that 500,000 people with burn injuries receive medical treatment each year; one-half of US burn victims are children, most with scald burns. This systematic study was initiated as follow-up to personal anecdotal evidence acquired over more than 10 years by MH Bhattacharyya regarding PO's efficacy for treating burns. The results demonstrate that PO counteracted dermal inflammation in both a mouse ear model of contact irritant-induced dermal inflammation and a second degree scald burn to the mouse paw. Furthermore, PO significantly counteracted the tactile allodynia and soft tissue injury caused by the scald burn. In mouse dorsal root ganglion neuronal cultures, PO added to the medium blocked adenosine triphosphate-activated, but not capsaicin-activated, pain pathways, demonstrating specificity. These results together support the hypothesis that a pine-oil-based treatment can be developed to provide effective in-home care for second degree burns.

Acetate and propionate impact on the methanogenesis of landfill leachate and the reduction of clogging components.

Synthetic leachate with different initial concentrations of acetate (500-2500 mg HAc/L) and propionate (500-3500 mg HPr/L) was treated with active biomass acclimated to landfill leachate under anaerobic mesophilic conditions for 72 h. Methanogenesis was observed within all samples during the first 48 h. The greatest removal of acetate (80-100%) and propionate (15-35%) was achieved in tests with initial concentrations ranging from 1500 to 190 0mg HAc/L and from 1000 to 1800mgHPr/L. Concurrent with the removal of acids, pH increased between 0.3 and 0.45 units, to above the threshold pH for precipitation of CaCO(3). Therefore, some 50-70% of dissolved Ca(2+) was removed from solution. This study suggests that by using an equalization tank (prior to the anaerobic digester) to maintain the acetate and propionate concentrations to within their optimum range would help to alleviate clogging through the removal of dissolved Ca(2+) and maximizing CH(4) production.

Characterization of quaternary ammonium oligomers by paired-ion reversed-phase liquid chromatography-mass spectrometry.

The separation of novel permanently charged oligomers was studied using paired-ion reversed-phase liquid chromatography. The polyionene studied is less than 5 kDa in size, but contains three oligomer series with different end-group chemistries. The complexity of this polyionene makes development of a single-dimension separation quite challenging. Separation under critical conditions was employed to fractionate the end-group conformations and then the chain length of the oligomers in each series was confirmed by LC-MS. The oligomers were then used to optimize a single-dimension HPLC separation. Precise modulation of the hydrophobicity of the ion-pair reagent and the stationary-phase chemistry yielded very high resolution one-dimensional separations.

Effect of airway disease on blood gas exchange in racehorses.

Inflammatory Airway Disease (IAD), exercise-induced pulmonary hemorrhage (EIPH), and upper airway obstruction (UAO) are common respiratory tract diseases that can decrease performance. The purpose of this retrospective study was to compare bronchoalveolar lavage fluid cytology and arterial blood gas analysis during a treadmill test by poorly performing racehorses presented to Purdue University. One hundred thirty-two horses with a history of poor performance were included in this study. Ten horses with no history or diagnosis of EIPH, IAD, or UAO served as controls. Horses were evaluated by rhinolaryngoscopy for upper airway abnormalities and underwent a standardized treadmill test, and samples were collected for blood gas analysis. Horses with IAD or EIPH had a more severe exercise-induced hypoxemia, (mean+/-SD; 84.8+/-1.5 and 86.0+/-1.7 mm Hg average Pao2, respectively), than horses in the control group (92.8+/-2.1 mm Hg). The average Pao2 of horses with only UAO (88.3+/-3.3 mm Hg) was not significantly different from control horses. Gas exchanges were the most severely impaired in horses affected with both EIPH and UAO because they exhibited the lowest Pao2 and highest Paco2 values (66.5+/-15.2 and 52.2+/-6.3 mm Hg, respectively).

An STS content map of human chromosome 11: localization of 910 YAC clones and 109 islands.

Physical mapping of human chromosomes at a resolution of 100 kb to 1 Mb will provide important reagents for gene identification and framework templates for ultimately determining the complete DNA sequence. Sequence-tagged site (STS) content mapping, coupled with large fragment cloning in yeast artificial chromosomes, provides an efficient mechanism for producing first-generation, low-resolution maps of human chromosomes. Previously, we produced a set of standardized STSs for human chromosome 11 regionally localized by fluorescence in situ hybridization or somatic cell hybrid analysis. In this paper, we used these as well as other STS content, and identify 109 islands spanning an estimated 218 Mb on the 126-Mb chromosome. Since about 62% of the islands contain markers ordered on chromosome 11 by genetic or radiation hybrid analysis, this data set represents a first-order approximation of a physical map of human chromosome 11. This set of clones, contigs, and associated STSs will provide the material for the production of a continuous overlapping set of YACs as well for high-resolution physical mapping based upon sampled and complete DNA sequencing.

Parathyroid hormone-related protein antigen localization distinguishes between mesothelioma and adenocarcinoma of the lung.

The distinction between pleural malignant mesothelioma and pulmonary adenocarcinoma remains a problem in diagnostic histopathology. Parathyroid hormone-related protein (PTHrP) has been demonstrated in the neoplastic cells of malignant mesotheliomata, using a polyclonal antiserum raised to synthetic PTHrP(1-16). In a series of 44 malignant mesotheliomata and 44 cases of pleural adenocarcinomata, PTHrP was localized immunohistochemically in 84 per cent of the mesotheliomata and in 11 per cent of the pleural adenocarcinomata. Normal and reactive mesothelium did not contain detectable PTHrP. The presence of PTHrP in a very high percentage of malignant mesotheliomata indicates the value of including it in the panel of antibodies utilized in the differential diagnosis of mesothelioma.

In situ hybridization of parathyroid hormone-related protein in normal skin, skin tumors, and gynecological cancers using digoxigenin-labeled probes and antibody enhancement.

We describe a novel procedure for in situ hybridization that combines the use of digoxigenin-labeled oligonucleotide probes with an antibody enhancement step that can be performed on formalin-fixed, paraffin-embedded tissues. Addition of a second antibody enhances the visibility of parathyroid hormone-related protein (PTHrP) mRNA expression from barely to highly discernible and interpretable, with virtually no nonspecific background expression. This technique has allowed visualization of PTHrP mRNA in normal human skin and epithelium-derived tumors. PTHrP mRNA expression was confined to the basal and spinous keratinocyte layers of skin. There was strong hybridization in the spinous keratinocyte layer and a low level of hybridization in the basal layer. An extensive panel of positive and negative controls included poly d(T) probe to indicate total mRNA present in the sections. Squamous cell carcinomas and basal cell carcinomas of the skin, from pathology archives, were examined for the presence of PTHrP mRNA. The results reflected previous immunohistochemical studies, with every squamous cell carcinoma hybridizing strongly with the PTHrP probes. The basal cell carcinomas showed no expression of PTHrP mRNA, although the total mRNA signal was very strong. The localization of PTHrP mRNA in the tumors of the gynecological tract also reflected the immunohistochemical findings, with expression found in the squamous cell carcinomas but not in the adenocarcinomas. In situ hybridization with digoxigenin-labeled oligonucleotide probes and antibody enhancement has provided a sensitive, highly specific procedure for detection of PTHrP mRNA in tumors and normal tissue.

Comparison of human and mouse T-cell receptor variable gene segment subfamilies.

Like the immunoglobulin Igh-V and Igk-V gene families, the human or mouse TCRV gene families may be grouped into subfamilies displaying > 75% nucleic acid sequence similarity among their members. Systematic interspecies sequence comparisons reveal that most mouse Tcr-V subfamilies exhibit clear homology to human TCRV subfamilies (> 60% amino acid sequence similarity). Homologous pairs of TCRV genes in mice and humans show higher sequence similarity than TCRV genes from different subfamilies within either species, indicating transspecies evolution of TCRV genes. Mouse and human homologues show conservation of their relative map order, particularly in the 3' region and a similar sequential and developmentally programmed expression. When the V regions from both species were analyzed together, local length differences and conserved residues in the loop regions were revealed, characteristic of each of the four TCRV families.

Human T-cell receptor variable gene segment families.

Multiple DNA and protein sequence alignments have been constructed for the human T-cell receptor alpha/delta, beta, and gamma (TCRA/D, B, and G) variable (V) gene segments. The traditional classification into subfamilies was confirmed using a much larger pool of sequences. For each sequence, a name was derived which complies with the standard nomenclature. The traditional numbering of V gene segments in the order of their discovery was continued and changed when in conflict with names of other segments. By discriminating between alleles at the same locus versus genes from different loci, we were able to reduce the number of more than 150 different TCRBV sequences in the database to a repertoire of only 47 functional TCRBV gene segments. An extension of this analysis to the over 100 TCRAV sequences results in a predicted repertoire of 42 functional TCRAV gene segments. Our alignment revealed two residues that distinguish between the highly homologous V delta and V alpha, one at a site that in VH contacts the constant region, the other at the interface between immunoglobulin VH and VL. This site may be responsible for restricted pairing between certain V delta and V gamma chains. On the other hand, V beta and V gamma appear to be related by the fact that their CDR2 length is increased by four residues as compared with that of V alpha/delta peptides.

Mouse T-cell receptor variable gene segment families.

All mouse T-cell receptor alpha/delta, beta, and gamma variable (Tcra/d-, b-, and g-V) gene segments were aligned to compare the sequences with one another, to group them into subfamilies, and to derive a name which complies with the standard nomenclature. It was necessary to change the names of some V gene segments because they conflicted with those of other segments. The traditional classification into subfamilies was re-evaluated using a much larger pool of sequences. In the mouse, most V gene segments can be grouped into subfamilies of closely related genes with significantly less similarity between different subfamilies.

Cholecystitis, cholelithiasis, and ganglioneuromatosis of the gall bladder: an unusual presentation of MEN type 2b.

A 40 year old man with multiple endocrine neoplasia type 2b (MEN 2b) presented with cholecystitis caused by gall stones. Twenty four years earlier, he had had a partial thyroidectomy for a cold nodule. At his initial presentation MEN 2b with medullary carcinoma of the thyroid had not been made. This was diagnosed while investigating his gall bladder symptoms and he was found to have asymptomatic residual medullary thyroid carcinoma and bilateral adrenal phaeochromocytomas. The cholecystectomy specimen contained several mixed calculi and extensive ganglioneuromatosis with large, prominent nerves containing ganglion cells in the gall bladder wall.

A sequence-tagged site map of human chromosome 11.

We report the construction of 370 sequence-tagged sites (STSs) that are detectable by PCR amplification under sets of standardized conditions and that have been regionally mapped to human chromosome 11. DNA sequences were determined by sequencing directly from cosmid templates using primers complementary to T3 and T7 promoters present in the cloning vector. Oligonucleotide PCR primers were predicted by computer and tested using a battery of genomic DNAs. Cosmids were regionally localized on chromosome 11 by using fluorescence in situ hybridization or by analyzing a somatic cell hybrid panel. Additional STSs corresponding to known genes and markers on chromosome 11 were also produced under the same series of standardized conditions. The resulting STSs provide uniform coverage of chromosome 11 with an average spacing of 340 kb. The DNA sequence determined for use in STS production corresponds to about 0.1% (116 kb) of chromosome 11 and has been analyzed for the presence of repetitive sequences, similarities to known genes and motifs, and possible exons. Computer analysis of this sequence has identified and therefore mapped at least eight new genes on chromosome 11.

Pseudocysts of the adrenal gland.

Three cases of pseudocysts or hemorrhagic cysts of the adrenal gland were encountered in female patients who presented with loin pain. The adrenalectomy specimens contained cystic structures; 2 had fibrous walls containing organizing blood clot, the remaining lesion was thin walled and contained blood stained fluid. Fat, normal adrenal cortical tissue and calcification were found within the cysts. These lesions (arising from vascular anomalies) require separation from hemorrhagic adrenal neoplasms. Awareness of adrenal pseudocysts and careful attention to the histological features aids this distinction.

Primary small cell carcinoma of the pancreas.

A 59 yr old male presented with abdominal pain and jaundice. CT scan showed a tumor in the pancreas with secondaries in the liver, peri-pancreatic lymph nodes, vertebral bone marrow and subcutaneous tissue of the neck together with a cervical lymph node. The patient died after 2 wks and autopsy revealed complete replacement of the pancreas by tumor. Careful dissection of both lungs failed to show any tumor. A diagnosis of primary small cell carcinoma of the pancreas was made. Review of the literature indicates that this type of carcinoma accounts for about 1% of all pancreatic cancers and is commoner in elderly males who have a history of cigarette smoking. There is usually a rapid progression of disease and widespread dissemination, especially if untreated. Some success has been obtained with chemotherapy.

Pigmented pheochromocytomas of the adrenal medulla.

Three primary pigmented pheochromocytomas of the adrenal gland are presented. The pigment in all cases proved to be melanin. Two of the pheochromocytomas were sporadic and histologically typical, except for a focal spindle cell configuration in one. It is believed that the morphologic appearance of these tumors represents divergent differentiation from neural crest, expressing typical pheochromocytoma (polygonal cells) and melanocytic features (melanin pigment).

Leiomyosarcoma of the thyroid: immunohistochemical and ultrastructural study.

A primary leiomyosarcoma of the thyroid was seen in a 54 yr old female. The patient had a solitary, cold nodule with no abnormalities of thyroid function tests. Histologically it was composed of characteristic interlacing fascicles of spindle cells which, electron microscopically, showed thin myofilaments with focal dense bodies. Immunohistochemistry showed a strong reaction with vimentin, smooth muscle actin and desmin. No epithelial differentiation was noted immunohistochemically or ultrastructurally. No tumor recurrence was detected 15 mths after surgery. It is postulated that the smooth muscle differentiation had arisen by metaplastic change or de-differentiation with re-differentiation of anaplastic thyroid carcinoma.