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Fluorescence microscopic studies on hemal lymph nodes in rats: a new immunobiological concept.

Hemal lymph nodes are characterized by a high content of blood cells most of them in different stages of erythrophagocytosis. These peculiar structures are not well understood up to now regarding their functional morphology. Above all, their biological relevance, especially to the phenomenon of disintegration of one's own blood cells, has eluded conclusive explanation so far. In the present study, hemal lymph nodes of 45 rats of the perirenal group were investigated by means of confocal laser fluorescence microscopy (CLSM) in combination with three fluorescent markers: latex standard particles, liposomes and autologous erythrocytes. Each marker briefly entered the hemal lymph nodes when injected into the kidney, whereas no notable migration took place after intravenous injection. Besides direct connections between hemal lymph nodes and the homolateral kidney, the study also revealed lymphatic communications with the contralateral kidney. Each marker was ingested by nodal macrophages, most of them surrounded by red blood cells (rosette formation) and laden with the by-products of cellular disintegration (erythrophagocytosis). Intimate contact of lymphocytes with macrophages as an expression of special interaction (emperipolesis) between both types of cells was frequently observed. A new concept is proposed, which ascribes to the hemal lymph nodes an important immunobiological role for the recognition of antigenic properties of one's own red blood cells permanently released by the kidney. The information macrophages obtain from these cells is presented to lymphocytes, which, in turn, initiates suppresser immune reactions. Under normal conditions, this mechanism of cellular identification and surveillance serves to preserve self-tolerance of the defense system against permanent renewal of one's own red blood cell population during a life time. In this way, an auto-aggressive immune anemia is circumvented.

Animals↗

[Lymphatic mapping and sentinel lymph node biopsy in gastric cancer].

INTRODUCTION: Lymphatic mapping and the sentinel lymph node (SLN) concept has been validated in malignant melanoma and breast cancer.However, the application for other solid tumors is still controversial. One of the most promising approaches is selective lymph node staging in gastric cancer.The presented pilot study evaluated the feasibility of the radiocolloid technique in gastric cancer patients and its value in predicting a positive nodal status. PATIENTS AND METHODS: Fifteen patients with gastric cancer (u T(1-3)) underwent endoscopic submucosal injection of 0.4 ml 60 MBq (99m)Tc-Nanocis around the tumor 17 (+/-3) h prior to surgery. After laparotomy the activity of all 16 (JGCA) lymph node stations was measured by a handheld probe. All patients underwent standard gastrectomy with systematic D2 lymphadenectomy. After resection the site was scanned for residual activity. All sentinel lymph nodes (SLN's) were removed ex vivo from the resected specimen and processed for intensified histopathologic assessment including serial sections and immunohistochemistry. RESULTS: In 14 of 15 patients at least one or more SLN's were obtained (93%), the median number of SLN's was 3 (1-5). Of the 14 patients, 9 revealed lymph node metastases. In eight of the nine patients the sentinel node(s) correctly predicted metastatic lymph node invasion. In five cases the lymph node station with positive sentinel node(s) was the only positive node station resulting in a sensitivity of 8/9 (89%). In one case immunohistochemical staining revealed micrometastases leading to an upstaging in 1/6 of the initially nodal-negative patients. CONCLUSION: Lymphatic mapping and sentinel node biopsy using the radiocolloid technique is feasible in gastric cancer. Limited results indicate a correct prediction of the nodal status and the potential of upstaging.Further studies seem to be justified to evaluate the clinical impact of the method.

Aged↗

Relationship between lymph node sinuses with blood and lymphatic metastasis of gastric cancer.

AIM: To elucidate the relationship between lymph node sinuses with blood and lymphatic metastasis of gastric cancer. METHODS: Routine autopsy was carried out in the randomly selected 102 patients (among them 100 patients died of various diseases, and 2 patients died of non-diseased reasons), their superficial lymph nodes locating in bilateral necks (include supraclavicle), axilla, inguina, thorax, and abdomen were sampled. Haematoxylin-Eosin staining was performed on 10 % formalin-fixed and paraffin-embedded lymph node tissue sections (5 um). The histological patterns of the lymph sinuses containing blood were observed under light microscope. The expression of CD31, a marker for endothelial cell, was detected both in blood and non-blood containing lymph node sinuses with the method of immunohistochemistry. RESULTS: Among the 1 322 lymph nodes sampled from the autopsies of 100 diseased cases, lymph node sinuses containing blood were found in 809 lymph nodes sampled from 91 cases, but couldn't be seen in the lymph nodes sampled from the non-diseased cases. According to histology, we divided the blood containing lymph node sinuses into five categories: vascular-opening sinus, blood-deficient sinus, erythrophago-sinus, blood-abundant sinus, vascular-formative sinus. Immunohistochemical findings showed that the expression of CD31 was strongly positive in vascular-formative sinuses and some vascular-opening sinuses while it was faint in blood-deficient sinuses, erythrophago-sinuses and some vascular-opening sinuses. It was almost negative in blood-abundant sinus and non-blood containing sinus. CONCLUSION: In the state of disease, the phenomenon of blood present in the lymph sinus is not uncommon. Blood could possibly enter into the lymph sinuses through the lymphaticovenous communications between the veins and the sinuses in the node. Lymph circulation and the blood circulation could communicate with each other in the lymph node sinuses. The skipping and distal lymphatic metastasis of gastric cancer may have some connection with the blood containing lymph node sinuses.

Adolescent↗

Procedure for lymph node dissection around splenic artery in proximal gastric cancer.

BACKGROUND/AIMS: The effect of lymph node metastasis around the splenic artery on the prognosis of proximal gastric cancer patients is not confirmed. The aim of this study is to clarify the optimal procedure for lymph node dissection along the splenic artery in proximal gastric cancer. METHODOLOGY: Proximal gastric cancer patients who underwent total gastrectomy with pancreaticosplenectomy were examined. The anatomical location of lymph nodes and the metastases around the pancreas were also studied in pancreatic cancer patients who underwent total pancreatectomy. RESULTS: Multivariate analysis of lymph node metastasis around the splenic artery showed that No. 11 lymph node metastasis was affected by No.10 lymph node that was predicted by depth of invasion. Multivariate analysis of prognostic variables by Cox's proportional hazard regression revealed that No. 10 lymph node metastasis was the significant factor affecting prognosis. No lymph node metastasis infiltrating the pancreatic parenchyma was observed in the pancreatic body or the tail. CONCLUSIONS: Total gastrectomy preserving the pancreas and spleen is the optimal procedure in proximal T2 gastric cancer. Total gastrectomy with splenectomy is appropriate in T3 cases, and distal pancreatectomy should be additionally done only in cases of direct invasion by the lymph node and/or the tumor to the pancreas.

Gastrectomy↗

Sentinel lymph nodes with technetium-99m colloidal rhenium sulfide in patients with esophageal carcinoma.

BACKGROUND: The authors assessed the detection of sentinel lymph nodes in patients with esophageal squamous cell carcinoma (SCC) using technetium-99m colloidal rhenium sulfide. They studied whether an analysis of sentinel lymph nodes using cytokeratin (CK) immunohistochemistry increased the accuracy of staging. METHODS: The authors observed 25 patients with thoracic esophageal carcinomas who underwent radical esophagectomy. The day before surgery, technetium-99m colloidal rhenium sulfide was injected into the submucosa at four sites around the primary tumor. Lymphoscintigraphy was performed. Esophagectomy and regional lymph node dissection were performed 17 hours after the technetium-99m injection. After surgery, the resected lymph nodes were evaluated by CK staining. RESULTS: Lymphoscintigraphy detected sentinel lymph nodes in 92% of the patients (23 of 25 patients). The accuracy of sentinel lymph node was 91.3% (21 of 23 patients), the sensitivity was 86.7% (13 of 15 patients), and the false-negative rate was 8.7% (2 of 23 patients). A comparison of the number of sentinel lymph nodes and clinicopathologic factors showed that there was a significant association between the number of sentinel lymph nodes and lymph node status (P < 0.01), pathologic stage (P < 0.05), and the number of metastatic lymph nodes (P < 0.05). Occult metastasis was detected by CK staining in 14 (56%) of the 25 patients and in 23 (1.7%) of 1406 lymph nodes. Because the 2 false-negative (sentinel lymph node-negative and nonsentinel lymph node-positive) patients who had occult metastases in the sentinel lymph nodes, the accuracy of sentinel lymph node evaluation using CK staining was 100% (23 of 23). CONCLUSIONS: Lymphatic mapping with technetium-99m colloidal rhenium sulfide was used to identify the lymphatic basin and was feasible in patients with esophageal SCC. An analysis of sentinel lymph nodes using CK immunohistochemistry increased the accuracy of sentinel lymph node.

Aged↗

Clinical evaluation of cutaneous malignant melanoma with histologically involved lymph node metastases.

Thirty-six cases of cutaneous malignant melanoma with histologically involved lymph nodes were studied during a 2-10 year study period, and the characteristics of the metastasized nodes and the associated prognosis were compared. The results were as follows: 1) the survival rate in cases with 3 or fewer (n = 16) metastasized nodes was significantly higher than in cases with 4 or more positive nodes (n = 20); 2) the survival rate in cases in which metastasized nodes were limited to one regional lymph node section (n = 12) was significantly higher than in cases where metastasis extended to two or more intra-regional sections of nodes (n = 24); 3) cases which had nodes measuring 3.00 cm or less (n = 25) had significantly higher survival rates than those with nodes of 3.01 cm or more (n = 11). Therefore, the results indicate that the number of metastasized lymph nodes, the extension into regional lymph node sections, and the size of the metastasized lymph node(s) can be considered as important prognostic factors for melanoma patients.

Follow-Up Studies↗

[Diagnosis and therapy of lymph node tuberculosis].

Due to an increase of lymph node tuberculosis and associated diagnostic problems the question for the easiest and most accurate diagnostic measures was asked. Furthermore, the success of surgical and pharmacologic treatment was evaluated. A total of 16 patients (nine females, seven males) treated at the medical outpatient clinic between 1977 and 1995 because of lymph node tuberculosis were evaluated retrospectively. Diagnosis had been established in all cases either by fine needle aspiration or surgical excision. The mean age was 31 years (range 17-68). In 10 patients cervical lymphomas were found. Other localisations were supraclavicular, inguinal, axillary and mediastinal. In 94% of the patients a fine needle aspiration was performed to ascertain diagnosis. Acid-fast bacilli were found in 36% and mycobacteria were cultured in 92%. Five patients (31%) had a surgical excision with acid-fast bacilli in two and positive cultures from all specimens. All patients had chemotherapy, in most cases started by a combination of three and followed by a combination of two drugs. According to our results a fine needle aspiration is justified as a first diagnostic step if lymph node tuberculosis is suspected. A positive result can be regarded as conclusive and spares surgical intervention. Since negative results do not exclude tuberculosis they justify surgical excision for final diagnosis.

Adolescent↗

Immunolocalization of peripheral lymph node addressins in normal and neoplastic human thymuses.

Peripheral lymph node addressin is a specific L-selectin ligand of the high endothelial venules that plays an important role in lymphocyte homing to lymph nodes. Tissue selective migration of lymphocytes through this pathway to the thymus has also been proposed. In this work, peripheral lymph node addressin expression was investigated immunohistochemically with a monoclonal antibody, clone MECA-79, in formaldehyde-fixed, paraffin-embedded tissue sections of 5 normal neonatal thymuses, 25 thymomas, 3 thymic carcinomas, and 2 thymic lymphoid hyperplasias. In normal thymuses, peripheral lymph node addressin expression was found in the endothelium of corticomedullary and medullary vessels surrounded by perivascular space. In type B thymomas and thymic lymphoid hyperplasias, peripheral lymph node addressin was detected in the vessels with perivascular spaces, at the medullary differentiation areas, and in paralymphoid follicles, respectively. However, in type A thymomas and thymic carcinomas, MECA-79-positive vessels were restricted to the remnants of pre-existing thymic tissue, and they were absent from the neoplastic areas. These findings suggest that in normal and most neoplastic thymuses, peripheral lymph node addressin is expressed by regions of vascular endothelium corresponding to postcapillary venules that may serve as a pathway for homing of recirculating lymphocytes to the thymus.

Adult↗

[Transbronchial needle aspiration under CT guide for enlarged mediastinal lymph node biopsy].

To ascertain the nature of enlarged mediastinal lymph nodes transbronchial needle aspiration (TBNA) was performed under CT guide through bronchoscopy. Forty-five enlarged mediastinal lymph nodes from 33 lung cancer patients were biopsied. The results indicated that 25 enlarged lymph nodes from 20 patients were due to cancer metastasis while 15 enlarged lymph nodes from 8 cases were due to inflammation, and those from the remaining 5 cases were normal. Therefore, about 33.3% of the enlarged mediastinal lymph nodes in lung cancer patients result from noncancerous inflammatory changes. TBNA under CT guide has the advantage of accurate localization, is easy to perform and safe.

Adenocarcinoma↗

Comparative activation requirements of human peripheral blood, spleen, and lymph node B cells.

Human peripheral blood, spleen, and lymph node B cells were stimulated with Cowan I Staphylococcus aureus (SA) or F(ab')2 fragments of anti-mu antibody (anti-mu) and various lymphokines and were analyzed for proliferation and generation of Ig-secreting cells (ISC). SA alone but not anti-mu stimulated minimal proliferation of each population. Recombinant IL 2 (r-IL 2) effectively promoted proliferation of SA-stimulated blood and spleen B cells, but supported less vigorous responses of lymph node B cells. By contrast, r-IL 2 enhanced DNA synthesis of all anti-mu-stimulated B cells early in culture, but did not promote sustained proliferation of anti-mu-stimulated lymph node B cells and only promoted ongoing DNA synthesis of some anti-mu-activated blood (eight out of 17) and spleen (five out of 14) B cell preparations. Recombinant interferon-gamma (r-IFN-gamma) and a commercial preparation of B cell growth factor (BCGF) also augmented DNA synthesis of all three B cell populations stimulated with SA or anti-mu early in culture, but neither alone was able to sustain maximal proliferation. Markedly enhanced sustained proliferation of all three anti-mu- and SA-stimulated B cell populations was noted when cultures were supported by the combination of r-IL 2 and BCGF, or to a lesser extent by r-IL 2 and r-IFN-gamma. The generation of ISC from SA-stimulated blood or spleen but not lymph node B cells was effectively supported by r-IL 2 alone. Differentiation of lymph node B cells required the combination of r-IL 2 and BCGF. These studies emphasize the importance of both the activation stimulus and the origin of the B cells in determining the lymphokine requirements of human B cell responsiveness.

B-Lymphocytes↗

Siderosis of lymph nodes patients with Hodgkin's disease.

Lymph nodes removed from 28 untreated patients with Hodgkin's disease all contained markedly increased amounts of hemosiderine, whether or not they were histologically involved in the disease. This finding was particularly striking in patients with the nodular sclerosis type of disease. Abnormal deposits of iron were also noted frequently in lymph nodes containing metastatic carcinoma, lymphoma of non-Hodgkin's type, and reactive hyperplasia, but in each case, with the exception of metastatic squamous cell carcinoma, the amount was significantly less than seen in Hodgkin's disease. The findings suggest that in patients with Hodgkin's disease and perhaps in those with other disorders in which abnormal tissue retention of iron underlies sideropenic anemia, lymph nodes are an important site of iron retention.

Adenocarcinoma↗

Common reactive erythrophagocytosis in axillary lymph nodes.

Erythrophagocytosis by histiocytes in the sinuses of axillary lymph nodes is a common yet little-known phenomenon. The axillary lymph node dissections of 23 patients were studied by light microscopy and graded for the amount of erythrophagocytosis. None of the patients had evidence of a systemic hemolytic process. Nineteen of them exhibited some degree of erythrophagocytosis, and this was present even in four of the six patients who never had a prior breast biopsy. Breast biopsy was associated with massive degrees of erythrophagocytosis in 8 of 17 patients, and after a postbiopsy interval of two weeks 11 of 13 patients had hemosiderin deposition in the lymph nodes, evidence of red blood cell breakdown. This study serves to substantiate statements, made by others without confirmatory data, that large degrees of erythrophagocytosis may be seen in axillary lymph nodes after breast biopsies and that small amounts of erythrophagocytosis may be identified in lymph nodes of patients with no prior trauma history.

Adult↗

Sentinel lymph node detection in canine oncological patients.

BACKGROUND: Sentinel lymph node detection was investigated in dogs with spontaneously occurring tumours. MATERIAL AND METHODS: In this pilot study, 24 client-owned spontaneously tumorous dogs presented for sentinel lymph node detection. A multiple method was used with a nuclear medicine technique (injection of 99mTc human serum albumin colloid) with scintigraphy and intraoperative guidance, and blue dye injection. RESULTS: Of the 35 lymph nodes histologically demonstrated to contain metastases, 34 (97%) were found by radioguided surgery, which means that one would have been missed in the intraoperative localisation process; 31 nodes (89%) were clearly visualised in the gamma camera images; only 27 (77%) were blue-stained by vital dye; a mere 8 lymph nodes (23%) were enlarged and therefore easily detectable by palpation. CONCLUSIONS: Data obtained from the harmless application of the sentinel node concept are useful for the radiopharmaceutist. The sentinel lymph node concept is well applicable in the veterinary clinic.

Journal Article↗

Sentinel lymph node detection by lymphoscintigraphy in malignant melanoma.

AIMS AND BACKGROUND: Sentinel lymph node (SLN) detection is currently employed in patients with malignant melanoma (MM) to spare them unnecessary lymph node dissection. METHODS AND STUDY DESIGN: We investigated 241 patients (130 men and 111 women, median age, 50 years (range, 14-92) with MM (192 before and 51 after surgical biopsy); two of them had more than one melanoma lesion. In each patient approx. 10 MBq of 99mTc Nanocoll in 0.1 mL (Nycomed Amersham Sorin; particle size range, 3-80 nm) was injected intradermally around the MM lesion or surgical scar. Dynamic acquisition was performed for 20 minutes (20 frames/min) and the study was concluded within four hours of injection. Using an external radioactive marker, the skin over the SLN was marked with China ink. RESULTS: 294 SLNs were scintigraphically identified: 117 in the inguinal region, 147 in the axillae, four in the submandibular region, three in the laterocervical region and 23 at other sites. In two patients no drainage was detected. In 43 patients more than one sentinel node was identified. In 13 patients with lesions located in the trunk the tracer drained towards multiple lymph node stations or unexpected lymph nodes (nine cases). Histology and immunohistochemistry diagnosed MM in 25 SLNs; 19 were positive for metastasis with hematoxylin-eosin staining, five with Hmb45 and one with CD68 immunostaining. All 25 detected lymphatic basins were excised. In nine of these basins there was metastatic involvement of at least one other lymph node besides the SLN. During follow-up which ranged from six to 86 months, metastatic disease was found in only one patient with a histologically negative SLN six months after surgery. CONCLUSIONS: This study confirms the utility of scintigraphic SLN detection in patients with MM. In most of the cases the procedure led the surgeon to evaluate the drainage area, which is unpredictable for lesions in the trunk and may be difficult to delineate using only patent blue dye. Furthermore, in approximately 10% of cases we observed dual drainage from individual lesions, mainly those located on the trunk. We will proceed to compare the results obtained during follow-up with those of an investigational group of patients with melanoma who were not subjected to lymphoscintigraphy for SLN detection in order to obtain well-founded information on the prognostic value of this technique.

Adolescent↗

Acute necrotizing lepromatous lymphadenitis: an erythema-nodosum-leprosum-like reaction in lymph nodes.

Histological examination of lymph-node biopsy specimens in 12 patients with erythema nodosum leprosum showed almost complete replacement of the node by lepromatous granuloma, together with considerable polymorph infiltration. Ziehl-Neelsen staining demonstrated numerous Mycobacterium leprae present in the nodes. The majority of these patients were very ill, and responded to prednisolone or corticotrophin.It is suggested that the histological appearances may represent an intensive inflammatory response in the lymph nodes followed by avascular aseptic necrosis.

Adrenocorticotropic Hormone↗

The suitability of hexyl cinnamic aldehyde as a calibrant for the murine local lymph node assay.

The murine local lymph node assay (LLNA) for the prospective identification of contact allergens assesses skin sensitization potential as a function of proliferative activity induced in lymph nodes draining the site of topical exposure to test chemical. This method has been endorsed recently as a stand alone test for the identification of contact allergens. We have now examined the suitability of hexyl cinnamic aldehyde (HCA), a recommended positive control for skin sensitization testing, as a calibrant for comparing the consistency of LLNA responses with time, and between laboratories, and thus for the routine assessment of assay reliability. Standard LLNAs were performed with CBA strain mice in 3 independent laboratories over a period of 8 years. Dose-response curves were used to derive mathematically the EC3 value (the estimated concentration of chemical necessary to cause a stimulation index (SI) of 3 compared with proliferation induced by concurrent vehicle controls). In each laboratory, 6 separate experiments were conducted using a single concentration of HCA (25%). Very similar stimulation indices were achieved, with mean values of 9.0, 6.5 and 6.6 recorded. A total of 10 dose-response experiments were performed independently in the 3 laboratories and these revealed that there was very little inter-laboratory, or temporal, variation in EC3 values. These data confirm that HCA responses in the LLNA are very stable and demonstrate that HCA provides a suitable calibrant for determining assay sensitivity and performance.

Acrolein↗

Molecular quantification and mapping of lymph-node micrometastases in cervical cancer.

BACKGROUND: A proportion of patients with cancer and lymph nodes negative on histology will develop recurrence. Reverse-transcriptase PCR (RT-PCR) is a highly sensitive method for detection of lymph-node micrometastases, but accurate quantitative assessment has been difficult. METHODS: We studied primary tumours and 156 lymph nodes from 32 patients with cervical cancer (stage IA2, IB1, and IB2) and 32 lymph nodes from nine patients with benign disease. A fully quantitative, real-time RT-PCR assay was used to document absolute copy numbers of the epithelial marker cytokeratin 19. Primers and probe were designed not to amplify either of the two cytokeratin 19 pseudogenes. FINDINGS: All primary tumours and histologically involved lymph nodes (six) had more than 106 copies of cytokeratin 19 mRNA per microg total RNA. Expression of cytokeratin 19 (up to 1.1 x 10(5) copies per microg RNA) was detected in 66 (44%) of 150 histologically uninvolved lymph nodes, and in nodes from 16 of 32 patients with cervical cancer. 15 of these 16 patients with evidence of micrometastases had the highest cytokeratin 19 transcription level in a first lymph-node drainage station (three obturator, six internal, and six external iliac node). Transcription of cytokeratin 19 was found at a low level in just one of 32 lymph nodes obtained from nine patients with benign disease. Median copy number of cytokeratin 19 transcription was significantly higher (>10(3) copies) in association with adverse prognostic features. INTERPRETATION: The results suggest that about 50% of early-stage cervical cancers shed tumour cells to the pelvic lymph nodes. The amount of cytokeratin 19 expression was related to clinicopathological features. Further studies are required to document the clinical implications of molecular micrometastases.

Adult↗

Lymph node metastases in carcinoma of the head of the pancreas region.

Histopathological examination of lymph node metastatic involvement in 139 specimens obtained from patients who underwent pancreatoduodenectomy or total pancreatectomy combined with wide resection of lymph nodes was performed, to clarify the critical areas of lymph node dissection in patients with carcinoma of the head of the pancreas region. Perigastric lymph node involvement in patients with carcinoma of the head of the pancreas was 14 per cent, in those with carcinoma of the distal bile duct 0 per cent and in those with carcinoma of the papilla of Vater 4 per cent. Para-aortic lymph node involvement in patients with carcinoma of the head of the pancreas, the distal bile duct and the papilla of Vater was 26, 9 and 0 per cent, respectively. On the basis of these results, pylorus-preserving pancreatoduodenectomy is indicated in almost all patients with carcinoma of the distal bile duct and the papilla of Vater. In patients with carcinoma of the head of the pancreas, however, wide dissection of lymph nodes, including para-aortic lymph nodes, should be carried out because of the relatively high incidence of para-aortic lymph node involvement.

Carcinoma, Ductal, Breast↗