Surgical outcomes of T4b oral cancers: assessment of prognostic factors and a need to re-evaluate the current staging system.

T4b oral cancer is a broad umbrella term for all advanced oral cancers, the prognosis of which varies drastically for disease of the same stage, according to the extent of the masticator space involvement. This was a retrospective observational study including all consecutive T4b oral squamous cell carcinoma patients treated surgically between January 2015 and January 2016 and followed up until January 2020. The disease was classified as upper disease or lower disease based on the anatomical location in relation to an imaginary plane passing through the base of the retromolar trigone. The prime objective was to evaluate overall survival and prognostic factors affecting overall survival. The projected 5-year overall and disease-free survival rates were 40.7% and 35.6%, respectively. The assessment of prognostic factors revealed that lower disease (lower anatomical subsites), bone invasion, and lymph nodal spread significantly affected survival. Patients with disease in an upper anatomical location without bone and nodal involvement can achieve fairly good survival (projected 5-year overall survival of 64.2%) when compared to the other subsets of patients. We propose a re-evaluation of the current staging system based on the prognostic features, so that all patients are not considered under a single stage, since their survival differs significantly.

Salvage surgery for recurrent carcinoma of the oral cavity: assessment of prognostic factors.

Salvage surgery is the most acceptable therapeutic option for disease control of loco-regional recurrences in oral cancers. Prognostic factors need to be assessed to select patients for salvage surgery who would benefit the most. This was a single-centre retrospective observational study conducted between 2015 and 2018. A total of 168 patients with recurrent oral cavity carcinoma who underwent salvage surgery were included for analysis. The primary endpoints of the study were to evaluate overall survival (rOS) after salvage surgery and prognostic factors affecting survival. In this study, the median rOS was 18 months and the median disease-free survival (rDFS) was 14 months. Advanced stage (hazard ratio (HR) 2.387, 95% confidence interval (CI) 1.496-3.808; P = 0.001) and multimodality treatment (HR 1.642, 95% CI 1.139-2.367; P = 0.008) in the initial disease, as well as nodal spread (HR 3.794, 95% CI 1.580-9.111; P = 0.008) and perineural invasion (HR 2.167, 95% CI 1.358-3.455; P = 0.001) in the recurrent disease, were found to adversely affect survival after salvage surgery. With thorough assessment of the prognostic factors and appropriate patient selection, survival may be favourable after salvage surgery for recurrent oral cavity carcinoma.

A proposed "Radiological Evaluation Score for Bone Tumors" (REST): An objective system for assessment of a radiograph in patients with suspected bone tumor.

BACKGROUND: Although radiographs have been widely used in the evaluation of patients with suspected bone tumors, the lack of an objective radiological assessment method leads to a challenge in reaching correct diagnosis. The study aimed to propose a Radiological Evaluation Score for Bone Tumors (REST) which includes eight radiological factors [characteristics, content, cortical breach, distinctiveness, distribution, periosteal reaction, fracture, and soft tissue swelling] to form a single score along with its validation by multidisciplinary observers. METHODS: We reviewed the radiographs of 100 patients with a primary bone tumor which were selected at random from the database between January 2017 and January 2019 of a tertiary cancer center. Four reviewers (two orthopedic oncologists and two surgical oncologists) independently assessed the radiographs, based on the reporting system of REST. We constituted two groups according to the probable diagnosis of bone tumor (suspected benign tumor and suspected malignant tumor). RESULTS: The mean score in the suspected benign tumor group was 1.1 (range 0-3, 95% CI 0.8-1.3) and in malignant tumor group was 6.1 (range 2-8, 95% CI 5.8-6.4). A receiver operator characteristic (ROC) curve for REST was with a cutoff of 3.5, with the most diagnostic value area under curve (AUC) of 0.99. The sensitivity was 98% and specificity was 100% with a positive predictive value of 100% and a negative predictive value of 98%. The inter-observer correlation coefficient was 0.985 (p value < 0.05), and Fleiss kappa value for the prediction of the benign or malignant lesion was 0.97 (p value < 0.05). The characteristics and content of tumor, cortical erosion, distinctiveness, distribution, periosteal reaction, and soft tissue mass had a significant correlation with the aggressiveness of bone lesion p value < 0.05. CONCLUSIONS: The Radiological Evaluation Score for Bone Tumors (REST) is a structured reporting and objective method for the assessment of radiographs in patients with suspected bone tumors. This method is a reliable and helpful tool for clinicians in their outdoor patient department to differentiate a radiograph of a suspected benign tumor from a malignant bone tumor.

A proposed ''A to Z RAM (Radiograph Assessment Method)'' for triage of patients with a suspected bone tumour.

INTRODUCTION: We propose a ''A to Z RAM (Radiograph Assessment Method)'' for evaluation of Radiograph of patients with a suspected bone tumour. METHODS: In the current study, ten radiological features with letters 'A, B, C, D, E, F and Z' were used and which included the age of the patient, involved part of the bone, characteristics, content, distinctiveness, the exterior of the bone, fracture, and zone of transition. Four independent observers (orthopaedic oncologists and surgical oncologists) evaluated a set of 30 radiographs of bone tumour selected at random from our hospital database based on A to Z RAM. We classified the lesions into two groups according to the traffic signal system; Green (suspected benign lesion) and Red (suspected malignant lesion). RESULTS: There were 18 (60%) benign bone lesions and 12 (40%) malignant lesions in the current study. 91.6% of malignant tumours and 88.8% of the benign tumours were identified correctly by the four observers. The inter-observer variability with Fleiss kappa was 0.884 (95% CI 0.7-1.03 p-value < 0.05), suggestive of agreement not by chance. These radiographs were again reassessed by the four observers after 3 months. The interobserver variability by Fleiss kappa was 1.0 (95% CI 0.8-1.1) suggesting complete agreement amongst the observers. Both orthopaedic oncologists had intra-observer kappa as 1.0 each and both surgical oncologists had 0.795 and 0.930 respectively. CONCLUSION: The proposed A to Z RAM is an easy to use and reproducible method for reviewing radiographs in the out-patient department along with clinical findings for better management of patients with suspected bone lesions. The A to Z RAM can be a medical triage tool and subdivide bone lesions into two subgroups i.e. suspected benign lesion with a suggestion of further investigations with MRI and biopsy and suspected malignant lesion with a suggestion of MRI or early referral to a tertiary cancer center with expertise in orthopaedic oncology. IMPLICATIONS FOR PRACTICE: The A to Z RAM (Radiologic Assessment Method) is a reproducible method for reviewing radiographs in the out-patient department and can be an aid for better management of patients. A to Z RAM is useful as a medical triage system, subdividing patients according to the probable diagnosis into a suspected benign lesion and suspected malignant lesion.

A high-throughput system for segmenting nuclei using multiscale techniques.

Automatic segmentation of cell nuclei is critical in several high-throughput cytometry applications whereas manual segmentation is laborious and irreproducible. One such emerging application is measuring the spatial organization (radial and relative distances) of fluorescence in situ hybridization (FISH) DNA sequences, where recent investigations strongly suggest a correlation between nonrandom arrangement of genes to carcinogenesis. Current automatic segmentation methods have varying performance in the presence of nonuniform illumination and clustering, and boundary accuracy is seldom assessed, which makes them suboptimal for this application. The authors propose a modular and model-based algorithm for extracting individual nuclei. It uses multiscale edge reconstruction for contrast stretching and edge enhancement as well as a multiscale entropy-based thresholding for handling nonuniform intensity variations. Nuclei are initially oversegmented and then merged based on area followed by automatic multistage classification into single nuclei and clustered nuclei. Estimation of input parameters and training of the classifiers is automatic. The algorithm was tested on 4,181 lymphoblast nuclei with varying degree of background nonuniformity and clustering. It extracted 3,515 individual nuclei and identified single nuclei and individual nuclei in clusters with 99.8 +/- 0.3% and 95.5 +/- 5.1% accuracy, respectively. Segmented boundaries of the individual nuclei were accurate when compared with manual segmentation with an average RMS deviation of 0.26 microm (approximately 2 pixels). The proposed segmentation method is efficient, robust, and accurate for segmenting individual nuclei from fluorescence images containing clustered and isolated nuclei. The algorithm allows complete automation and facilitates reproducible and unbiased spatial analysis of DNA sequences.

Acute effects of centrophenoxine short term administration on counting behavior in rats.

This experiment assessed the effects of Centrophenoxine on counting behavior in rats. Intraperitoneal Centrophenoxine injections were given to rats before training on a 20-fixed-consecutive-number schedule of reinforcement. The primary measure was the number of consecutive lever presses. After Centrophenoxine injections, the number of consecutive lever presses was lower relative to baseline measures. The measures of variability indicated that although the consecutive number of responses was lower this could not be accounted for in terms of run length variability. The behavioral pattern produced by the injections was discussed in terms of possible shortterm physiological effects that affect performance.

Dietary restriction: effects on radial maze learning and lipofuscin pigment deposition in the hippocampus and frontal cortex.

Previous studies have shown increased life span and decreased lipofuscin deposition in brain structures when both premature and young adult animals are calorically restricted of an otherwise nutritionally adequate diet. Three-months-old C57BL/6 mice were subjected to 12 months of caloric restriction (2 g/day), and subsequently exposed to a radial maze learning paradigm. Mice in the diet restriction group showed faster learning and higher asymptotic performance on the radial maze task, as well as lower lipofuscin deposition in the neurons of hippocampus and frontal cortex relative to control mice fed ad libitum. The results suggest that dietary restriction has effects on radial maze learning, and this improved behavioral performance was associated with significant reduction in lipofuscin pigment deposition in the neurons of hippocampus and frontal cortex.

Learning deficits occur in young mice following transfer of immunity from senescent mice.

The extent to which immune processes contribute to senescence-related neurological/behavioral impairment was examined using an adoptive transfer procedure. C57BL/6 mice aged 22 to 24 months showed impaired ability for acquisition of an active avoidance response when compared with younger mice aged 3 months. An immunofluorescence assay of the sera of these mice indicated that only sera from the senescent mice reacted with brain antigen. When tested three months following irradiation and receipt of bone marrow/spleen cell suspensions from senescent mice, young mice showed senescence-like serum-brain reactivity and declines in their abilities to acquire learning. Young control mice receiving cell suspensions from age-matched donors showed no evidence of serum-brain reactivity or learning deficits, suggesting that impaired learning was related to acquisition of aged immunity and not a nonspecific effect of the transfer procedure. These findings indicate that immune processes may be involved in the etiology of senescence-related neurological/behavioral dysfunctions.

Correlation between a learning disorder and elevated brain-reactive antibodies in aged C57BL/6 and young NZB mice.

Previous studies have indicated an increase in brain-reactive antibodies (BRA) in sera of aging mammals and an autoimmune disorder underlying senescence has been suggested. Since New Zealand Black (NZB) mice have a shorter lifespan and greater propensity for autoimmune diseases than C57BL/6 mice, various age groups from both strains of mice were investigated for simultaneous occurrence of BRA serum titer and deficits in learning. NZB mice exhibited a marked learning deficit as well as higher BRA levels at all ages. C57BL/6 mice showed increased BRA and a learning deficit only at advanced ages. The findings of "precocious" BRA titers along with marked learning deficits, both occurring at young ages in NZB mice and both similar to defects seen in the normal mice at senescence and in patients with senile-dementia, suggest that NZB mice may serve as a useful animal model of pre-senile dementia.

Immune manipulations and brain-reactive antibody formation in aging mice.

Previous studies have demonstrated an increase in brain-reactive antibodies (BRA) in sera of various mammals as a function of age. The present study attempts to obtain a better understanding of the mode of formation of BRA by using various immunologic manipulations in young and old C57BL/6 mice. The mean BRA levels were significantly higher in older mice than in the young ones. Mixtures of bone marrow cells and spleen cells from old donors were able to adoptively transfer high BRA levels in young irradiated mice and this was not altered by pretreatment of the cells with anti-Thy-1.2 or anti-Ig serum plus complement. Transfer of young cells to old irradiated hosts did not lower the high BRA levels in the hosts. However, when the old hosts were also thymectomized and grafted with neonatal thymuses, BRA levels were significantly lowered. The studies tend to indicate that thymic involution or loss of thymic function in aging plays an important role in BRA formation in sera of aged mice.

Effects of centrophenoxine on lipofuscin in the retinal pigment epithelium of old mice.

The effects of centrophenoxine on the retinal pigment epithelium (RPE) of 17 month old female mice have been studied. Animals were injected subcutaneously for 3 months (60 injections) with the drug (0.1 mg/g of body wt) daily in 0.1 M phosphate buffered saline at pH 7.0. The morphological changes in the pigment layers of the retina of both eyes were studied by light and electron microscopy and the lipofuscin pigment was demonstrated by its autofluorescence and ultrastructural characteristics. There was a significant reduction of the lipofuscin pigment in the treated animals, but the melanin pigment remained unchanged. The lipofuscin granules also appeared less osmiophilic and showed a greater preponderance of membranes and vacuoles. Although the precise mechanism of action of the drug is not clear, an increased protective function of the pigment epithelium by the drug has been suggested.

Effects of controlled dietary restriction on brain-reactive antibodies in sera of aging mice.

Previous studies have demonstrated that underfeeding in both premature and young mature animals may extend the life span as well as preserve the functions of the immune system. The effects of caloric restriction for a period of 12 months on different organs as well as the formation of brain-reactive antibodies in young mature mice (3 months) were tested. These animals showed a significantly lower weight of the total body and various organs including the brain, spleen, adrenals and kidneys. The brain weight/body weight ratio, on the other hand, was significantly higher in these mice. Sera in the dietary animals were mostly negative while those of control animals of the same age and sex had high levels. The present study supports the earlier observations that controlled dietary restriction is able to slow down the age-related deterioration of the immune system. The inhibition of brain-reactive antibody formation in these animals might also be related to a delayed onset of autoimmune disorders.

Senescence related changes in brain diazepam binding and motor performance.

Senescent mice showed attenuation of habituation in locomotor activity and marked deficit in the motor performance requiring limb coordination. Also, the binding of 3H-diazepam to the crude synaptosomal fraction prepared from cerebral cortex of the senescent mice was significantly altered. The binding maximum (Bmax) was significantly greater in the aged (23-26 months) mice, but no change was found in the apparent dissociation constant (Kd). Senescent related changes appear to include neurochemical alteration of the endogenous benzodiazepine system (increased receptor sites without any change in receptor affinity) and, perhaps, related physiological deficits in the limb coordination and/or strength.

Brain-reactive antibodies in sera of aging non-human primates.

Previous studies in our laboratory demonstrated an age-related increase in the brain-reactive antibodies in sera of aging mice. The present paper deals with the study of brain-reactive antibodies in sera of non-human primates (Macaca nemestrina) of three age groups (4, 10 and 20 years) by an indirect immunofluorescence method. The results indicated a progressive increase in the serum levels of these antibodies with advancing age. Since all three age groups examined demonstrated brain-reactive antibodies in sera, the age of onset of these antibodies could not be determined in these animals. It has been suggested that the formation of these antibodies could be evidence of autoimmune reactions which might play a significant role in neuronal degeneration during aging.

Morphological changes in the cerebellar cortex of aging Macaca nemestrina.

The cerebellar cortices in 4, 10 and 20 year of Macaca nemestrina have been examined for the number of Purkinje (P) and granule cells and the deposition of lipofuscin in P cells in relation to aging. Lipofuscin distribution significantly increased with in the P cells in these animals. The number of P cells was significantly reduced, while there were no changes in the number of granule cells. It appears from this and other studies that the Purkinje cells are more prone to aging changes than the granule cells of the cerebellum both in lipofuscin formation and cell loss. Although the precise functional significance of these changes in P cells is not clear, their vulnerability may be related to changes in motor function in old age.

Lipofuscinogenesis in mice early treated with centrophenoxine.

Previous studies in our and other laboratories indicated that there is a reduction in the neuronal lipofuscin in old rodents after several weeks of treatment with centrophenoxine. The present study investigates whether this chemical can prevent pigment formation if given early in life before the onset of pigmentogenesis. The study shows that the drug did not stop lipofuscin formation in 1 month old mice. But there was a consistent decrease in the pigment in the neurons of cerebral cortex and hippocampus of the treated animals compared to the age-matched controls. The degree of reduction was largely dependent on the duration of the treatment and a significant diminution was noted after treatment for five months or more.

Centrophenoxine: effects on aging mammalian brain.

A study was made of the effects of centrophenoxine on the learning and memory of old mice. The results were correlated with changes in neuronal lipofuscin in the cerebral cortex and hippocampus. Old female mice (11-12 months) were treated with centropheoxine for three months and their learning and memory were tested in a T-maze. The number of trials required to attain the criterion in the 20 treated old mice were compared with those for 20 untreated mice of the same age and for 20 younger untreated mice. The treated animals learned the task with significantly fewer trials, and also exhibited a reduction of neuronal lipofuscin pigment in both the cerebral cortex and the hippocampus. The changes in lipofuscin were demonstrated by study of the characteristic autofluorescence, and by histolchemical and ultrastructural (electron microscope) observations.

Effects of dihydroergotoxine mesylate on aging neurons in vitro.

C1300 mouse neuroblastoma cells gradually accumulate lipofuscin-like pigment when they are maintained in culture. Pigment was demonstrated by positive straining for acid phosphatase and with periodic acid-Schiff stain. Pigment was formation in cells was reduced by exposure of the cells to lower doses of dihydroergotoxine mesylate which also induced neurite formation and increased protein synthesis. Since lipofuscin appears to originate as a result of wear and tear within the cells, the drug probably exerts its beneficial effects by reducing the rate of intracellular wear and tear associated with aging.