Effectiveness of radiation therapy on brain invasion by human papillomavirus-related multiphenotypic sinonasal carcinoma: A case report.

Human papillomavirus (HPV)-related multiphenotypic sinonasal carcinoma (HMSC) is newly suggested and characterized by HPV-related tumors. HMSC has a relatively good prognosis. No cases of brain invasion have been reported to date. We encountered a case of brain invasion by HMSC, in which we assessed the effectiveness of radiotherapy in comparison with biopsy and autopsy. A 69-year-old man was referred to a hospital three months after intracerebral hemorrhage (ICH). Contrast magnetic resonance imaging revealed a tumor in the ethmoid sinus involving the brain. We performed transnasal biopsy and intensity-modulated radiotherapy for sinonasal and intracranial lesions. Despite radiotherapy, the patient died on day 41 after radiation. Biopsy specimens displayed mixed findings of epithelial and mesenchymal components. The tumor was immunoreactive for p16, and the RNA in situ hybridization for HPV was positive. Finally, we diagnosed the patient as having HMSC. Autopsy of the sinonasal tissue revealed a reduction in the number of tumor cells. There was a marked reduction in the number of tumor cells in the sinonasal tissue compared to that in the invaded brain tissue. The effectiveness of radiotherapy could depend on the histopathological components and location of the lesion, even in the same patient.

Association between cancer immunity and treatment results in uterine cervical cancer patients treated with radiotherapy.

OBJECTIVE: To evaluate proteins related to tumor immune response and treatment outcome from radiotherapy for uterine cervical cancer patients. METHODS: We performed a retrospective immunohistochemical staining of 81 patients with uterine cervical cancer who underwent definitive radiotherapy. We examined the expression of programmed death ligand 1, human leukocyte antigen class I, tumor-infiltrating CD8+, and forkhead box P3+ (FoxP3+) T cells in tumor tissues. RESULTS: In biopsy specimen, patients with a higher number of CD8+ T cells and FoxP3+ T cells had a better disease-specific survival than patients with a lower number of CD8+ T cells and FoxP3+ cells (P = 0.018 and P = 0.009). Multivariate analysis showed that equivalent dose in 2 Gy fractions (EQD2) of the minimum dose to 90% of the high-risk clinical target volume, FoxP3+ T cells and expression of human leukocyte antigen class I were significant prognostic factors. When the EQD2 is 70 Gy or more, a higher local control rate is obtained regardless of the number of CD8- or FoxP3-positive cells. When EQD2 is <70 Gy, the number of CD8-positive cells has a significant impact on treatment outcome: the recurrence rate (local recurrence rate + distant metastasis rate) was 46.2% in the group with a CD8 value of 230 or higher, whereas the recurrence rate was 75.7% in the group with a CD8 value of less than 230. CONCLUSION: The combination of CD8 or FoxP3 with EQD2 can be potentially useful to predict the treatment results of radiotherapy for cervical cancer, leading to individualized optimal selection of treatment for cervical cancer.

Microscopic observation of chemical modification in sections using scanning acoustic microscopy.

A scanning acoustic microscope (SAM) calculates the speed of sound (SOS) through tissues and plots the data on the screen to form images. Hard tissues result in greater SOS; based on these differences in tissue properties regarding SOS, SAM can provide data on tissue elasticity. The present study evaluated whether tissue modifications, such as formalin fixation, periodic acid-Schiff (PAS) reactions and protein degradation, changed the acoustic properties of the tissues and whether SAM could be a useful tool for following chemical changes in sections. The fixation process was observable by the increased SOS. During the PAS reaction, the glycosylation of tissues was characterized by an increased SOS. Mucous or glycogen distribution was visualized and was found to be statistically comparable among lesions and states. Protease digestion by pepsin led to a decreased SOS. Tissue sensitivity to proteases varied due to the stage, cause and duration of inflammation or ageing. Changes in acoustic properties were more sensitive than those in optical histology. SAM facilitates the visualisation of the time course or distribution of chemical modifications in tissue sections, thus aiding their comparison among tissues. SAM may be an effective tool for studying changes such as protein cross-linkage, tissue repair and ageing.

Observations of amyloid breakdown by proteases over time using scanning acoustic microscopy.

Amyloid consists of insoluble beta-fibrillar proteins with stable structures. The Congo red staining method for histologically detecting amyloid is unsuitable for quantitatively assessing amyloid fibers. Scanning acoustic microscopy (SAM) detects the attenuation of sound (AOS) through sections. This study aimed to clarify whether AOS values reflected the amount of amyloid fibril degradation in tissues. Formalin-fixed paraffin-embedded unstained sections of various types of amyloidosis were digested with different endopeptidases. The AOS images after digestion were observed over time via SAM. The corresponding Congo red-stained images were followed to identify the amyloid. The amyloid and nonamyloid portions were statistically examined over time to determine the changes in the AOS values. Most of the amyloid areas showed significantly different AOS values from nonamyloid portions before digestion and significantly decreased after digestion; these findings corresponded with the disappearance and waning of the Congo red staining in the light microscopic images. Some nonamyloid areas with high AOS masked the reduction in AOS in the amyloid areas. The method used in this study may help detect the amyloid quantity and determine the appropriate treatment method for removing amyloid deposits from tissues.

Pulmonary imaging with a scanning acoustic microscope discriminates speed-of-sound and shows structural characteristics of disease.

Tissue elasticity can be detected using a scanning acoustic microscope (SAM), whereby acoustic images are created from the speed of sound through tissues. This system discriminated pulmonary tissue components and demonstrated distinct acoustic images of the lung; these results corresponded well to those obtained using the conventional microscope. SAM provides the following benefits: (1) images are acquired in only few minutes without requirement for staining, (2) basic data is obtained for low-frequency ultrasonic examination, and (3) speed of sound from each lesion is digital and comparable among diseases. Comparative analysis of cancer invasion, post-inflammatory fibrosis, and deposition disease was possible using the data obtained with the system, and the results showed good correlation with those using the conventional microscope and by clinical diagnosis. The SAM system is applicable not only to pulmonary diseases but also to various diseases in other organs.

Double neuroendocrine ductal carcinomas in situ coexisting with a background of diffuse idiopathic neuroendocrine cell hyperplasia of breast: a case report and hypothesis of neuroendocrine tumor development.

This article reports the case of a 72-year-old woman with two nodules of neuroendocrine (NE) ductal carcinoma in situ coexistent with a background of NE cell hyperplasia. Both tumors, 15 and 3 mm in size, were incidentally revealed on computed tomography without any apparent clinical symptoms. The tumors showed similar histological features, and more than 50% of the tumor cells patchily expressed NE markers, such as chromogranin A, synaptophysin, CD56, and somatostatin receptor type 2. The surrounding nontumor ductal cells also showed spotty or linear positivity for NE markers in contrast to the cells of normal atrophic breasts, which rarely present with NE cells. Moreover, focal mucin production was also observed in the peripheral ducts. It is hypothesized that idiopathic breast NE cell hyperplasia with multiple small nests of NE cells may extend to form a true mass of NE neoplasms.

Autopsy cases of fulminant bacterial infection in adults: clinical onset depends on the virulence of bacteria and patient immune status.

To assist physicians in recognizing the potentially fatal onset of symptoms in cases of fulminant bacterial infection, we analyzed 11 autopsy cases of such infection (four caused by Streptococcus pneumoniae, four by S. pyogenes, one by S. dysgalactiae subsp. equisimilis, one by Staphylococcus aureus, and one by Vibrio vulnificus). Clinicohistopathologic features were evaluated. All patients experienced sudden onset of hypotension and multiple organ failure, leading to unexpected death. Blood culture confirmed bacteremia. The main chief complaints were gastrointestinal symptoms (45%) and limb pain (36%). All had an underlying chronic illness (82%), e.g., a hematologic disorder (36.3%) or liver cirrhosis (27.2%). Necrotizing fasciitis occurred in only 55% of cases, with none involving pneumococcal infection. Laboratory tests typically showed C-reactive protein elevation but without leukocytosis, indicating a high-level inflammatory state. In ten cases, death was attributed to circulatory collapse due to sepsis; severe pulmonary congestion and hemorrhage were present in these cases. The onset of fulminant bacterial infection depends on both virulence of the bacterium and status of the host defense system.

Stiffness reduction and collagenase resistance of aging lungs measured using scanning acoustic microscopy.

Lung tissue stiffness is altered with aging. Quantitatively evaluating lung function is difficult using a light microscope (LM) alone. Scanning acoustic microscope (SAM) calculates the speed-of-sound (SOS) using sections to obtain histological images by plotting SOS values on the screen. As SOS is positively correlated with stiffness, SAM has a superior characteristic of simultaneously evaluating tissue stiffness and structure. SOS images of healthy bronchioles, arterioles, and alveoli were compared among young, middle-aged, and old lung sections. Formalin-fixed, paraffin-embedded (FFPE) sections consistently exhibited relatively higher SOS values than fresh-frozen sections, indicating that FFPE became stiffer but retained the relative stiffness reflecting fresh samples. All lung components exhibited gradually declining SOS values with aging and were associated with structural alterations such as loss of smooth muscles, collagen, and elastic fibers. Moreover, reaction to collagenase digestion resulted in decreased SOS values. SOS values of all components were significantly reduced in young and middle-aged groups, whereas no significant reduction was observed in the old group. Protease damage in the absence of regeneration or loss of elastic components was present in old lungs, which exbited dilated bronchioles and alveoli. Aging lungs gradually lose stiffness with decreasing structural components without exposure to specific insults such as inflammation.

Scanning acoustic microscopy imaging of cellular structural and mechanical alterations from external stimuli.

Cells incur structural and functional damage from external stimuli. Under scanning acoustic microscopy (SAM), speed of sound (SOS), attenuation, and thickness values are plotted to visualize cellular stiffness, viscosity, and size. The obtained digital data are then compared using statistical analysis. In the present study, we aimed to investigate the alterations in the mechanical and structural characteristics of cancer cells in response to anticancer drugs, acidic fluids, and microwave burdens using SAM. We found that active untreated cells showed increased thickness and reduced SOS and attenuation, whereas dying treated cells displayed reduced thickness and increased SOS. Tannic and acetic acid treatments and microwave irradiation all increased SOS and attenuation and reduced thickness, which meant that these treatments made cells thinner, stiffer, and more viscous. Furthermore, the different anticancer drugs interacted with cancer cells to induce characteristic changes in SAM values. These structural and mechanical alterations induced in cells were difficult to observe under light microscopy. However, under SAM, cancer cell activity and function corresponded consistently with changes in SAM values. Cellular damage parameters were statistically compared between the different treatments, and time-dependent cellular changes were established. SAM observation can therefore reliably evaluate cancer cell damage and recovery after chemotherapy and physical therapy. These results may help evaluate the therapeutic efficacy of various treatments.

A case of intravenous leiomyomatosis with high levels of hyaluronan.

Intravenous leiomyomatosis (IVL) is a rare benign tumor. The clinical behavior can be life-threatening due to extension through the pelvic veins. A 70-year-old woman was referred to our hospital with IVL originating from a uterine leiomyoma and extending to the inferior vena cava. The patient was diagnosed on the basis of the results of various studies, and the tumor was resected completely through a single-stage approach. The intravascular tumor was 20 cm long, multinodular and rubbery. Microscopic findings showed benign smooth muscle that was partly hyalinized and fibrous. Immunohistochemical studies revealed that hyaluronan was expressed more prominently in IVL than in uterine leiomyomas. IVL has viscoelastic properties and contains a large amount of hyaluronan, which may promote invasion during pathogenesis.

Tunica intima compensation for reduced stiffness of the tunica media in aging renal arteries as measured with scanning acoustic microscopy.

OBJECTIVES: Aging causes stiffness and decreased function of the renal artery (RA). Histological study with light microscopy can reveal microscopic structural remodeling but no functional changes. The present study aimed to clarify the association between structural and functional aging of the RA through the use of scanning acoustic microscopy. METHODS: Formalin-fixed, paraffin-embedded cross-sections of renal arteries from 64 autopsy cases were examined. Speed-of-sound (SOS) values of three layers, which correspond to the stiffness, were compared among different age groups. SOS of the tunica media was examined in terms of blood pressure (BP) and SOS of the ascending aorta. Vulnerability to proteases was assessed by SOS reduction after collagenase treatment. RESULTS: The tunica intima presented inward hypertrophy with luminal narrowing, and the tunica media showed outward hypertrophic remodeling with aging. SOS of the tunica media and internal and external elastic laminae showed a reverse correlation with age. SOS of the tunica media was negatively correlated with BP and strongly associated with that of the aorta. The tunica media of young RAs were more sensitive to collagenase compared with the old ones. CONCLUSIONS: Scanning acoustic microscopy is useful for observing the aging process of the RA. This technique simultaneously shows structural and mechanical information from each portion of the RA. In the process of aging, the RA loses contractile function and elasticity as a result of protease digestion. The tunica media and the internal and external elastic laminae exhibit reduced stiffness, but the tunica intima stiffens with atherosclerosis. As a consequence, the RA's outer shape changes from round to oval with inward and outward hypertrophy. This indicates that the inner resistant intima supports the mechanical weakness of the tunica media to compensate for an increase in BP with aging.

Retrospective DVH analysis of point A based intracavitary brachytherapy for uterine cervical cancer.

Combining external beam radiotherapy (EBRT) with intracavitary brachytherapy (ICBT) is important for definitive treatment of cervical cancer. In cervical cancer patients receiving radiotherapy, we evaluated treatment outcomes in relation to dose-volume histogram parameters, including the computed tomography (CT)-based high-risk clinical target volume (HR-CTV) for ICBT. Between 2010 and 2015, 89 consecutive cervical cancer patients were mostly treated with 40 Gy of EBRT in 20 fractions and 18 Gy of ICBT prescribed to point A in 3 fractions. CT scans were obtained during ICBT. The HR-CTV D90 was calculated and the total doses of ICBT and EBRT were converted to the equivalent dose in 2 Gy fractions (EQD2). When the patients were divided into four groups according to EQD2 of the HR-CTV D90, the 3-year local recurrence-free survival rates were 95.2, 78.4, 52.7 and 42.9% for patients receiving >80 , 70-80 , 60-70 and <60 Gy, respectively. There was a significant negative correlation between EQD2 of the HR-CTV D90 and the HR-CTV volume at first ICBT (r = -0.713). Local recurrence was more frequent when the HR-CTV volume was ≥22 cc and EQD2 of the HR-CTV D90 was <70 Gy. Multivariate analysis showed that EQD2 of the HR-CTV D90 and concurrent chemotherapy (≥4 cycles) were significant determinants of overall survival. HR-CTV D90 was an important prognostic indicator for local recurrence. HR-CTV D90 >70 Gy is required for the better local control, especially in patients with a larger HR-CTV (≥22 cc at initial ICBT).

Tuberous sclerosis patient with neuroendocrine carcinoma of the esophagogastric junction: A case report.

BACKGROUND: Tuberous sclerosis complex (TSC) is a rare inherited disease with non-cancerous tumor growths in the skin, brain, kidneys, heart, and lungs. The co-occurrence of neuroendocrine neoplasm (NEN) with TSC is even rarer. There have been few reports on the relationship between TSC and neuroendocrine tumors (NETs), and fewer on the relationship between TSC and neuroendocrine carcinoma (NEC), a subtype of NEN. This is the first reported case of NEC occurring at the esophagogastric junction in a patient with TSC. CASE SUMMARY: A 46-year-old woman visiting our hospital for the treatment of TSC was admitted to the emergency department with tarry stools and dizziness. Computed tomography scans revealed thickness of the gastric cardia, multiple metastatic lesions of the liver, and enlarged lymph nodes near the lesser curvature of the stomach. Esophagogastroduodenoscopy revealed a type 3 tumor located from the esophagogastric junction to the fundus, and the pathological diagnosis by biopsy was NEC. The patient was treated with seven courses of cisplatin + irinotecan, followed by eight courses of ramucirumab + nab-paclitaxel, one course of nivolumab, and two courses of S-1 + oxaliplatin. Twenty-three months after the first treatment, the patient died because of disease progression and deterioration of the general condition. CONCLUSION: This case of NEC occurring in a patient with TSC indicates a difference in the occurrence of NETs and NECs.

BK virus nephropathy: clinical experience in a university hospital in Japan.

OBJECTIVES: To review the medical records of patients with BK virus nephropathy (BKVN) following kidney transplantation in our institution. METHODS: We screened patients for decoy cells using urine cytology, assessed serum creatinine levels, and conducted a graft biopsy, as well as assessed the presence of plasma BK virus DNA by quantitative real-time polymerase chain reaction. The treatment of BKVN was based on the decreased use of immunosuppressants. RESULTS: Overall, six male patients were studied (mean age 40.8 years, range 18-58; mean donor age 45.2 years, range 15-67). A positive urine cytology screen led to the subsequent detection of plasma BK virus DNA in the five patients with urine cytology results positive for decoy cells. In the four patients in whom plasma BK virus DNA was detected, a maximum value of DNA of > or = 10 000 copies/mL was observed. Time elapsed from transplantation to BKVN diagnosis ranged from 3 to 62 months. Although the two cadaver grafts were lost, the loss was not due to any effects directly associated with BKVN. The other four grafts are still functioning with a mean creatinine level of 1.8 mg/dL. Most of the patients with BKVN were regarded as being in a state of heightened immunosuppression. BK virus transition to blood was prevented in one patient. CONCLUSIONS: Early diagnosis of BKV infection with reduction of immunosuppression may potentially counter BK viremia and retard progression of BKV nephropathy. Decoy cell screening by urine cytology as well as plasma BK virus DNA screening should be considered in addition to the required graft biopsy in kidney transplant recipients, particularly in those with impaired graft function.

Patellar impingement by synovial cord following anterior cruciate ligament reconstruction.

We found a cordlike structure (synovial cord) in the patellofemoral joint causing patellar impingement in a 45-year-old man after anterior cruciate ligament (ACL) reconstruction. Arthroscopic examination revealed that a cylindrical cordlike soft tissue with a smooth surface extended between the medial and lateral capsule of the patellofemoral joint. The tissue histologically consisted of dense collagen fibers in parallel bundles covered with thin layers of synovial tissue; in addition, there was a proliferation of small vessel patches surrounded by hemosiderin-laden macrophages, which indicated bleeding episodes due to impingement in the patellofemoral joint. To our knowledge, this is the first case report of a synovial cord in the patellofemoral joint causing patellar impingement following ACL reconstruction.

Acoustic Histology with Specific Dyes and Antibodies.

The present study aims to identify specific staining methods for acoustic histology. We compared attenuation-of-sound (AOS) images from scanning acoustic microscopy (SAM) with light microscopy (LM) images. Ethanol-fixed tissue or cytology samples and formalin-fixed surgical or autopsy specimens were examined. Nuclei, collagen, elastic fibers and polysaccharides and various antigens, including cell surface, cytoplasmic, nuclear and stromal substances, were observed. Samples with various fixation methods were used. Hematoxylin staining had significantly higher AOS values in accordance with staining duration. Specific staining for collagen, elastic fibers and polysaccharides increased the AOS values of the specific substance. Using diaminobenzidine tetrahydrochloride in NiCl2 solution as a substrate for horseradish peroxidase increased the AOS values to those suitable for acoustic immunostaining. Collagenase digestion after collagen staining decreased AOS values, reflecting collagen density and distribution. Staining with specific dyes or acoustic immunostaining enabled the histologic localization of specific substances by SAM, similar to LM.

Mechanical Weakness of Thoracic Aorta Related to Aging or Dissection Predicted by Speed of Sound with Collagenase.

Scanning acoustic microscopy reveals information on histology and speed of sound (SOS) through tissues. Slower SOS corresponds to lower stiffness. The aim of the present study was to investigate whether SOS values reflect the degree of degeneration with aging or dissection and whether enzymatic digestion susceptibility is distinct. The SOSs of media other than the atheromatous areas of normal and surgical dissections were measured and compared using medial degeneration grade (MDG) scores. To evaluate the damage rate, SOS was assessed after collagenase digestion. SOS scores negatively correlated with aging and MDG scores. Dissected aortas had higher SOS and MDG scores without age correlation. Collagenase digestion was present in all aortas, but older aortas were more injured than younger aortas. Dissected aortas were more vulnerable to collagenase. Older and dissected aortas expressed specific extracellular matrix components to compensate for mechanical weakness. The present method can evaluate mechanical weakness corresponding to histology to investigate the cause of rupture.

Phase 2 Study of Neoadjuvant Treatment of Sequential S-1-Based Concurrent Chemoradiation Therapy Followed by Systemic Chemotherapy with Gemcitabine for Borderline Resectable Pancreatic Adenocarcinoma (HOPS-BR 01).

PURPOSE: Preoperative treatment is recommended for borderline resectable pancreatic ductal adenocarcinoma. However, the standard treatment has not yet been determined. We conducted a multicenter phase 2 study to investigate the efficacy of neoadjuvant treatment of sequential chemoradiation followed by chemotherapy. METHODS AND MATERIALS: All enrolled patients were treated by preoperative chemoradiation (a total dose of 50.4 Gy in 28 fractions and orally administered S-1 at 80 mg/m2 on the day of irradiation) followed by chemotherapy (administration of gemcitabine at 1000 mg/m2/dose on days 1, 8, and 15 in 3 cycles of 4 weeks) and attempted curative resection. The primary outcome was an R0 resection rate among patients who completed preoperative treatment and pancreatectomy. The threshold of the R0 resection rate was defined as 74% based on a previous study of up-front surgery. RESULTS: Forty-five patients were included. Twenty-one patients could not undergo pancreatectomy because of progressive diseases (n = 14), adverse events (n = 5), or consent withdrawal (n = 2), and 4 patients underwent additional resection after dropping out. The resection rates were 53.3% and 62.2% in the per-protocol set (PPS) and full analysis set (FAS) populations, respectively. The R0 resection rates were 95.8% (95% confidence interval, 78.9%-99.9%) and 96.4% (81.7%-99.9%) in the PPS and FAS populations, respectively. The median overall survival and progression-free survival of all the included patients were 17.3 and 10.5 months, respectively. The median survival time of the patients with pancreatectomy was significantly longer than that of the patients without pancreatectomy in the PPS (27.9 vs 12.3 months; P = .001) and FAS populations (32.2 vs 11.8 months; P < .001). CONCLUSIONS: This study revealed that a long duration of preoperative treatment of sequential chemoradiation followed by systemic chemotherapy provides a high rate of R0 resection and sufficient survival time in patients undergoing pancreatectomy.