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Modification of lymph by lymph nodes. III. Effect of increased lymph hydrostatic pressure.

Previous studies have shown that lymph nodes function as fluid exchange chambers in which the protein concentration of lymph is changed in the direction required to establish equilibrium of the Starling forces acting across the nodal blood-lymph barrier. We examined the effect of increased lymph hydrostatic pressure on efferent lymph by use of an isolated dog popliteal node preparation in which lymph having a protein concentration averaging 27.6 +/- 1.2% (SD) of that of plasma was infused into the node at a flow rate averaging 45.6 +/- 0.2 (SD) microliter/min. We compared steady-state values of prenodal and postnodal lymph flow and protein concentration following step increases in efferent lymph pressure from 0 to over 15 mmHg. Increasing efferent lymph pressure to values less than about 8 mmHg caused the efferent lymph protein concentration to increase; however, further increases in lymph pressure caused the lymph protein concentration to decrease to values approaching those attained at very low lymph pressures. We suggest that the failure of high lymph pressure to increase lymph protein concentration might be caused by blood vessel collapse within the node, a condition believed to increase nodal blood capillary pressure and to decrease blood-lymph barrier filtration coefficient. An important finding was that increasing efferent lymph pressure caused significant amounts of lymph proteins to be lost during nodal transit. Therefore, it appears that increasing efferent lymph pressure to very high values has little effect on lymph protein concentration but has great effect on postnodal lymph protein flux.

Animals↗

Modification of lymph by lymph nodes. II. Effect of increased lymph node venous blood pressure.

A previous study from this laboratory demonstrated that lymph nodes can change the protein concentration and colloid osmotic pressure of lymph by transfer of protein-free fluid between the blood and lymph compartments. In that study a Starling force disequilibrium across the blood-lymph barrier caused fluid to transfer through the barrier in the direction required to establish equilibrium of Starling forces. In the present study we examined the effect of increased lymph node venous blood pressure on efferent lymph protein concentration and efferent lymph flow. We utilized an isolated dog popliteal lymph node preparation in which afferent lymph having various protein concentrations was perfused into the node at an average flow rate of 19.1 +/- 0.3 (SD) microliter/min. We compared steady-state values of prenodal and postnodal lymph flows and protein concentrations during various steady-state levels of lymph node venous blood pressure. When venous pressure was increased, the protein concentration of the efferent lymph invariably decreased and the efferent flow rate invariably increased. Measurements showed that an average of 96% of the change in lymph protein concentration was caused by transfer of protein-free fluid through the lymph node blood-lymph barrier. The results of this study indicate again that the lymph node functions as a fluid exchange chamber in which fluid is transferred between the blood and lymph compartments in the direction required to establish equilibrium of the Starling forces across the blood-lymph barrier.

Animals↗

Number and distribution of pelvic lymph nodes and effect of surgical pathologic factors on pelvic lymph node status in patients with early-stage cervical carcinoma treated with radical hysterectomy and pelvic lymph node dissection.

PURPOSE OF INVESTIGATION: To report the number and distribution of pelvic lymph nodes and to identify surgical pathologic factors that best predict positive pelvic lymph nodes in patients with early-stage cervical carcinoma treated with radical hysterectomy and pelvic lymph node dissection (RHND). METHODS: Data from the files of 126 patients with cervical carcinoma treated by RHND at the Soroka Medical Center from 1962 through 2005 were analyzed. RESULTS: The status of pelvic lymph nodes was known in 114 patients. The exact number of lymph nodes removed from the pelvis of each patient was known in 111 patients. The mean number of lymph nodes removed from the pelvis per patient was 26.6 (median 23; range 1-62). Positive pelvic lymph nodes were found in 35 (30.7%) of the patients with a mean of 3.4 (median 2; range, 1-15) positive pelvic lymph nodes per patient. In a univariate analysis, positive lymph vascular space invasion and positive parametrial and/or paracervical involvement were significant predictors of positive pelvic lymph nodes, whereas penetration > or = 50% of the thickness of the cervical wall and grade 2+3 were of borderline significance. In a multivariate analysis, positive lymph vascular space invasion was the strongest and the only significant predictor of positive pelvic lymph nodes, whereas positive parametrial and/or paracervical involvement was of borderline significance. CONCLUSIONS: In patients with early-stage cervical carcinoma treated with RHND, positive lymph vascular space invasion emerged to be the strongest and most significant predictor of positive pelvic lymph nodes.

Analysis of Variance↗

Thoracic duct lymph and PEEP studies in anaesthetized dogs. I. Lymph formation and the effect of a thoracic duct fistula on lymph flow.

The effect of positive end-expiratory pressure ventilation (PEEP, 11-12 mmHg, 60-90 min without, 19 h with circulatory support) on fractional escape rate of plasma proteins (FER), and on thoracic duct lymph flow draining against jugular venous (LFJVP) or atmospheric pressure (LFAP) was studied in anaesthetized dogs. FER was 10.8%/h, 15.3%/h, and 8.5%/h before, during, and after PEEP, respectively, indicating augmented lymph formation probably due to the increase in venous pressure from 4.8 to 10.8 mmHg during PEEP. LFJVP was 39 microliter/min per kg before PEEP, decreased transiently during PEEP but the steady state value (up to 19 h) was not different from control, and increased transiently after PEEP. LFAP was 37, 80, and 38 microliter/min per kg before, during, and after PEEP, respectively. Long-term PEEP increased LFAP fourfold. Changing the drainage mode during PEEP yielded an immediate increase from LFJVP = 34 to LFAP = 79 microliter/min per kg and an instantaneous reduction from LFAP = 95 to LFJVP = 35 microliter/min per kg. Lymph protein concentration and protein lymph/plasma ratio increased concomitantly with LFAP during PEEP suggesting augmented hepatic contribution to LFAP, augmented intestinal contribution was revealed by labelling intestinal lymph using olive oil orally, muscular lymph flow was not increased as shown by i.m. Evans blue. In conclusion, the augmentation of venous pressure by PEEP promotes capillary filtration but obstructs lymph drainage from the thoracic duct into the jugular vein. PEEP imbalances formation and return of lymph and affects the development and removal of oedema.

Animals↗

Minimal risk of macrometastases in the non-sentinel axillary lymph nodes in breast cancer patients with micrometastatic sentinel lymph nodes and preoperatively ultrasonically uninvolved axillary lymph nodes.

Micrometastases in the sentinel lymph node (SLN) carry a considerable risk of macrometastases in the non-sentinel lymph nodes (NSLN), resulting in axillary lymph node dissection (ALND). Preoperative ultrasound (US) examination of the axillary lymph nodes combined with a fine-needle aspiration biopsy (FNAB) has been proved to discover metastases in the axillary lymph nodes. The aim of our study was to assess the risk of macrometastases in NSLN in patients with micrometastatic SLN after a preoperative US examination of the axillary lymph nodes. The study included 36 patients in whom, after preoperative axillary US, micrometastases in the SLN were revealed and ALND was subsequently performed. At final histopathology, no macrometastases were discovered in the NSLN. In four patients, additional micrometastases were discovered in the NSLN. In conclusion, the risk of macrometastases in the NSLN in patients with preoperatively ultrasonically uninvolved axillary lymph nodes is minimal.

Adult↗

Sampling of lymph from lymph vessels afferent to the supramammary lymph gland in the cow.

A technique for catheterization of a lymph vessel afferent to the supramammary lymph gland in the cow is described. The operation was made with the cow under general anaesthesia. The lymph vessel was catheterized with a 30 cm long heparinized polyethylene catheter with an external diameter of 2.5 to 3 mm. Generally at least 5 ml of lymph could be collected within a period of 3 to 5 minutes. In 11 out of 15 cows lymph could be collected for 7 days or longer.

Animals↗

Cutaneous melanoma: therapeutic lymph node and elective lymph node dissections, lymphatic mapping, and sentinel lymph node biopsy.

Early clinical observation in cancer patients suggested that tumors spread in a methodical, stepwise fashion from the primary site, to the regional lymphatics, and only then to distant locations. Based on these observations, the regional lymphatics were believed to be mechanical barriers, at least temporarily preventing the widespread dissemination of tumor. Despite evidence now available disputing its validity, this barrier theory has guided the surgical management of the regional lymphatics in cancer patients for more than a century, influencing the use of such surgical modalities as therapeutic lymph node dissection, elective lymph node dissection, and most recently lymphatic mapping and sentinel lymph node biopsy. No published randomized controlled trial exists that demonstrates improved overall patient survival for cancer of any type, including melanoma, after surgical excision of regional lymphatics. This article will review the biology of lymphatics as it relates to regional tumor metastasis, and based on available information, offer practical recommendations for the clinical dermatologist and their patients who have cutaneous melanoma.

Animals↗

Afferent lymph and lymph borne cells: their influence on lymph node function.

In AO rats the afferent lymphatics to the right cervical lymph nodes (LN) were interrupted and the LN were encased in silicone rubber tubes to prevent reunion of the lymphatics. At regular intervals over the next 12 weeks the following were measured in comparison with the intact contralateral LN - LN weight, influx of lymphocytes from the blood, blood flow, the incorporation of 125IUdR and the incorporation of 35S-sulphate into high endothelial venules (HEV). Systematic histological observations are also reported. One day after deafferentization lymphocyte influx was significantly reduced although blood flow was unchanged and a temporary increase in LN weight was associated with crowding of the lymphatic sinuses with small lymphocytes. The subsequent decline in lymphocyte influx was biphasic and quicker than the decline of other parameters--being undetectable by 6 weeks. Flattening of HEV and diminished secretion of 35S-sulphate was noted at 1 week and progressive degeneration and eventual disappearance of the HEV network was seen by 6-12 weeks. Doubtlessly because of lack of antigenic stimulation 125IUdR incorporation, and numbers of lymphoblasts, plasma cells and finally germinal centres were progressively reduced. The numbers of macrophages and interdigitating cells (IDC) were greatly reduced by 3 weeks and very few were present at 6 weeks probably because most or all arrive in afferent lymph and have a limited life span in the LN. At 12 weeks the LN was difficult to recognize as such since only stromal cells and occasional small lymphocytes remained. In supplementary experiments u.v. irradiation of the LN at the time of deafferentization reduced lymphocyte influx without affecting blood flow suggesting that a u.v. sensitive cell like the IDC may influence lymphocyte influx. In conclusion the involution of the deafferentized LN is partly due to the lack of antigen but progression to the complete loss of specialized structure and function is probably due to lack of other factors including non-lymphoid cells that normally arrive in afferent lymph.

Animals↗

Lymph node mapping and sentinel lymph node biopsy for evaluation of axillary lymph node status in early invasive breast cancer. Our experience.

The Authors show their preliminary experience with the sentinel lymph node biopsy (SLNB) in clinical early invasive breast cancer (T1N0). During a period of 15 months, forty-two patients were submitted to SLNB upon Tc99-colloid albumin injection and SLN identification by lymphoscintigraphy. The middle number of lymph nodes found in the SLNB was 1 (1-3), whereas the middle number of lymph nodes identified in level I/II ALND specimens was 15. The SLN was identified with success in all cases (100%). The axilla was positive for metastasis in 4/42 cases. The SLN was positive in all four cases in which nodal metastasis was identified. The negative predictive value of SLN was 100%. The SLN was the only site of metastasis in 3/4 cases. The SLN pathological status accurately reflected the lymphatic basin status, but further investigation is needed to define the optimal timing of colloid injection and method of examination of the SLN.

Adult↗

Sentinel lymph node biopsy alone without axillary lymph node dissection--follow up of sentinel lymph node negative breast cancer patients.

AIMS: To evaluate the rate of axillary recurrences in sentinel lymph node (SLN) negative breast cancer patients after sentinel lymph node biopsy (SLNB) alone without further axillary lymph node dissection (ALND). METHODS: Between May 1999 and February 2001 all patients who had primary invasive breast cancer and were SLN negative were eligible for this prospective study. SLNB was performed by using the combined method with radioactive tracer and blue dye. SLNs were examined by frozen section, standard H/E staining and immunohistochemistry staining. SLN negative patients did not receive further ALND. Follow-up was done three-monthly with clinical controls, blood samples and ultrasound of the breast and axilla. An annual mammogram was performed. RESULTS: 116 patients with T1 or T2 invasive breast cancer were included in this trial. All 116 patients had negative SLNs in frozen sections, in H/E staining and in immunohistochemistry staining. The mean number of removed SLNs was 2.03+/-1.22. Mean tumor size was 17.15+/-7.62 mm. Postmenopausal patients totalled 79.3 and 20.7% of patients were premenopausal. No local or axillary recurrences occurred at a mean duration of follow-up of 22.12+/-6.38 months. CONCLUSION: The absence of axillary recurrences after SLNB without ALND in SLN negative breast cancer patients supports the hypothesis that SLNB is accurate and safe while providing less surgical morbidity. Short term results are very promising. SLNB without ALND in SLN negative patients is an excellent procedure for axillary staging in a cohort of breast cancer patients with small tumors.

Adult↗

Predicting regional lymph node metastasis in carcinoma of the penis: a comparison between fine-needle aspiration cytology, sentinel lymph node biopsy and medial inguinal lymph node biopsy.

OBJECTIVE: To evaluate the accuracy of clinical examination and fine-needle aspiration cytology (FNAC) in detecting groin metastases in patients with carcinoma of the penis, and to assess the positive and negative predictive value (PPV, NPV) of a preliminary sentinel lymph-node biopsy (SNB) and biopsy of the most medial of the horizontal group of inguinal lymph nodes (MIN) in selecting patients for an ilio-inguinal block dissection. PATIENTS AND METHODS: The study comprised 28 patients (56 groins) with Stage I (one), Stage II (11) and Stage III (16) carcinoma of the penis. All patients underwent a detailed clinical examination followed by FNAC of the palpable inguinal nodes, and were subsequently submitted for block dissection. The MIN, the SN and the rest of the inguinal and iliac nodes were histologically examined separately for metastases. RESULTS: The clinical evaluation had a sensitivity of 74%, a specificity of 61%, a PPV of 57% and a NPV of 77%. The corresponding values for FNAC were all 100%, and the specificity and PPV for both MIN and SN were 100%. The sensitivity and NPV of MIN were higher than for SN, although not significantly so. CONCLUSION: Clinical examination alone is inaccurate in selecting patients with carcinoma of the penis for block dissection. FNAC is accurate and specific when nodes are palpable; in those with impalpable nodes a preliminary MIN biopsy followed by SNB if the MIN biopsy is negative will accurately select all patients with metastases in the groin nodes. This can be performed by examining frozen sections of the lymph nodes; if positive, block dissection can be carried out at the same time.

Adult↗

Cost-analysis of staging methods for lymph nodes in patients with prostate cancer: MRI with a lymph node-specific contrast agent compared to pelvic lymph node dissection or CT.

The aim of this study was to compare the costs of three strategies in patients with prostate cancer in a specific setting: firstly, a strategy including MR lymphography (MRL) in which pelvic lymph node dissection (PLND) is foregone in case of a negative result. The second strategy involves computed tomography (CT) followed by a biopsy or PLND. The third strategy consists of PLND without imaging beforehand. A decision analytic model was constructed. This model represented the diagnostic process for patients with prostate cancer and intermediate or high risk for nodal metastases, comparing the costs of the three strategies. Cost analysis was done from the health care perspective. The model indicated that the expected costs for the MRL strategy were 2,527 euro. The expected costs for the strategy using CT were 3,837 euro and for PLND 3,994 euro. These results show that potential savings performing MRL instead of CT were 1,310 euro and 1,467 euro for PLND. Sensitivity analyses show that variation in costs of PLND was most influential on the costs of all strategies. However, the overall savings pattern did not alter. Average costs of MRL staging in our institution are less than for CT and PLND in staging lymph nodes of patients with prostate cancer and who are intermediate or high risk for nodal metastases.

Biomarkers, Tumor↗

The architecture of rat lymph nodes. III. The lymph nodes and lymph-borne cells of the congenitally athymic nude rat (rnu).

The lymph nodes (LN) of the congenitally athymic nude rat(rnu) were compared with the LN of non-nude littermates. The mesenteric and coeliac LN were smaller in the rnu rats, but the axillary and the cervical LN were larger. Germinal centres were found in the LN of nude rats. They were scarce and often very small, but some were of normal structure, especially in the cervical LN. The endothelium of post-capillary venules in the LN or normal rats, was seen. The paracortex was extremely depleted of lymphocytes but remained a distinct area occupied predominantly by pale interdigitating cells. In some LN the medullary sinuses were distended and the cords engorged with plasma cells. The predominant cells in thoracic duct lymph were immunoglobulin-positive B lymphocytes. The hourly output of these cells was the same as that in non-nude littermates. However, small numbers of thoracic duct cells were positive for the markers identified by the monoclonal antibodies W3/13 and W3/25, although in normal rats these are not expressed on small B lymphocytes.

Animals↗

Risk factors for patients with pelvic lymph node metastases following radical cystectomy with en bloc pelvic lymphadenectomy: concept of lymph node density.

PURPOSE: We evaluated the clinical outcomes and risk factors for progression in a large cohort of patients with lymph node metastases following en bloc radical cystectomy and bilateral pelvic lymphadenectomy. MATERIALS AND METHODS: From July 1971 through December 1997, 1,054 patients underwent radical cystectomy and bilateral pelvic-iliac lymphadenectomy for high grade, invasive transitional cell carcinoma of the bladder. Of these patients 244 (23%) with a median age of 66 years (range 36 to 90) had pathological lymph node metastases. Overall 139 of the 244 patients (57%) received some form of chemotherapy. At a median followup of greater than 10 years (range 0 to 28) outcomes data were analyzed in univariate analysis according to tumor grade, carcinoma in situ, primary bladder tumor stage, pathological subgroups, total number of lymph nodes removed and involved with tumor, and lymph node density (total number of positive lymph nodes/total number removed). In addition, the form of urinary diversion and the administration of chemotherapy were also evaluated. Multivariate analysis was then performed to analyze these variables independently. RESULTS: The incidence of positive lymph nodes increased with higher p stage and pathological subgroups. Of 669 patients 75 (11%) with organ confined primary tumors and 169 of 385 (44%) with extravesical tumor extension had involved lymph nodes. The median number of lymph nodes removed in the 244 lymph node positive cases was 30 (range 1 to 96), while the median number of positive lymph nodes was 2 (range 1 to 63). Overall recurrence-free survival at 5 and 10 years for the 244 patients with lymph node positive disease was 35% and 34%, respectively. Patients with lymph node positive disease and an organ confined primary bladder tumor had significantly improved 10-year recurrence-free survival compared with those with extravesical tumor extension (44% vs 30%, p = 0.003). The total number of lymph nodes removed at surgery was also prognostic. Patients with 15 or less lymph nodes removed had 25% 10-year recurrence-free survival compared with 36% when greater than 15 lymph nodes were removed. Recurrence-free survival at 10 years for patients with 8 or less positive lymph nodes was significantly higher than in those with greater than 8 positive lymph nodes (40% vs 10%, p <0.001). The novel concept of lymph node density was also a significant prognostic factor. Patients with a lymph node density of 20% or less had 43% 10-year recurrence-free survival compared with only 17% survival at 10 years when lymph node density was greater than 20% (p <0.001). On multivariate analysis the total number of lymph nodes involved, pathological subgroups of the primary bladder tumor, lymph node density and adjuvant chemotherapy remained significant and independent risk factors for recurrence-free and overall survival. CONCLUSIONS: Patients with lymph node tumor involvement following radical cystectomy may be stratified into high risk groups based on the primary bladder tumor, pathological subgroup, number of lymph nodes removed and total number of lymph nodes involved. Lymph node density, which is a novel prognostic indicator, may better stratify lymph node positive cases because this concept collectively accounts for the total number of positive lymph nodes (tumor burden) and the total number of lymph nodes removed (extent of lymphadenectomy). Future staging systems and the application of adjuvant therapies in clinical trials should consider applying lymph node density to help standardize this high risk group of patients following radical cystectomy.

Adult↗

T and B lymphocyte populations in human normal lymph node, regional tumour lymph node and inflammatory lymph node.

We have determined the T and B lymphocyte subpopulations in normal human lymph nodes. The lymphocyte profile was the same irrespective of the anatomical distribution and was similar to that found in peripheral blood with identical T and B cell values but with a lower Fc and a higher C3-receptor-bearing lymphocyte subpopulation. This pattern showed a marked change in the regional nodes of patients with mammary carcinoma and nodes draining a variety of other solid tumours, with a fall in T and pronounced elevation of B, Fc and C3 cells but with a persistence of C3 predominance. The lymphocyte profile found in tonsils and nodes draining inflammatory foci was a similar but further exaggeration of the tumour node pattern, with reversal of T and B cell ratios. The T and B lymphocyte percentages in the peripheral blood of patients with clinically localized breast cancer are identical to those of the healthy controls. Different Fc/C3 subsets exist in peripheral blood and lymphoid structures and probably represent a differential functional heterogeneity. Proximity of tumour to the draining node modified this profile.

B-Lymphocytes↗

Killer cell (K) activity in human normal lymph node, regional tumour lymph node and inflammatory lymph node.

Human normal lymph nodes, irrespective of their anatomical site of origin, have a low K cell activity, which may not be detected except with the appropriate target cell and at high lymphocyte to target cell ratios (100:1). This very low killer cell activity is also found in all the homolateral axillary nodes of patients with clinical stage I and II carcinoma of the breast and in the regional draining nodes of a variety of solid tumours, whether small and localized or large and with extensive spread. In all cases proximity to the tumour and obious hyperplastic changes in the nodes have no modifying effect. This pattern of minimal reactivity is similarly found with tonsillar lymphocytes and with nodes draining inflammatory foci. The Fc and C3 receptors on surface membranes are dectected with ease, and pretreatment of lymphocytes by incubation, washing and enzymatic treatment fail to alter their reactivity, thus excluding effector cell inhibition by immune complexes. The killer cell activity of lymphocytes from the blood of breast tumor patients is similar to the activity of lymphocytes from healthy controls.

Binding Sites↗

Quantitation of changes in lymph protein concentration during lymph node transit.

Many investigators assume the protein concentration and colloid osmotic pressure of interstitial fluid and lymph to be identical even after the lymph has passed through a lymph node. We quantitated the degree of modification of lymph by the dog popliteal lymph node by perfusing isolated lymph nodes in situ at physiological flow rates with homologous plasma or plasma diluted to low protein concentration. This enabled us to compare directly prenodal and postnodal lymph flows and protein concentrations. When undiluted plasma was infused into the node, fluid filtered from the blood into the lymph, diluting the lymph. When diluted plasma was infused, fluid was absorbed from the lymph, concentrating the lymph. Nearly all (98%) of the change in lymph protein concentration could be explained by transfer of protein-free fluid either into or out of the lymph. However, when the nodes were perfused with lymph having a colloid osmotic pressure that exactly balanced the hydrostatic and osmotic forces acting across the lymph node blood-lymph barrier, the lymph was not modified during nodal transit. This "equilibrium colloid osmotic pressure" averaged 60% of that of plasma. The concentrating-diluting mechanism became more significant as the perfusion rate decreased and/or as the colloid osmotic pressure of the afferent lymph was made progressively greater than or less than the equilibrium colloid osmotic pressure. We conclude that lymph nodes modify lymph protein concentration and colloid osmotic pressure except when these are already at equilibrium values for given lymph node conditions. Therefore, the assumption that postnodal lymph is representative of interstitial fluid, especially at low but still physiological lymph flows, is likely to be incorrect.

Animals↗