Some problems in the laboratory findings in multiple myeloma

The clinical usefulness of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) levels in serum and pathogenetic mechanism of hypoalbuminemia and hypocholesterolemia in multiple myeloma (MM) were investigated. In cases of MM with a history of pathological fracture, the level of serum ALP was significantly higher than normal. Thus, elevated ALP in MM patients may be an indicator of the occurrence of a pathological fracture within the past 2 months. The levels of serum LDH in about 80% of the MM patients were within normal limits despite the presence of a malignant tumor. These patients showed a normal pattern of isoenzymes and more mature types according to the Greipp classification. In contrasts, the patients with elevated serum levels of LDH showed the tumor pattern of the isoenzymes and the plasmablastic type. The total cholesterol concentration was correlated with the total protein levels and the serum cholinesterase. These findings were the same as those in patients with nephrotic syndrome and polyclonal hypergammaglobulinemia without liver dysfunction. These results suggest that the decreased cholesterol in MM is due to a reduction in the synthesis of albumin in the liver.     

Chemical inhibition method for alkaline phosphatase isoenzymes in human serum.

The authors adapted a chemical inhibition procedure using L-phenylalanine and urea as specific inhibitors to quantitate the activities of bone, liver, and intestinal alkaline phosphatase (ALP) isoenzymes in human serum. The results of this assay were compared with electrophoretic separation, gamma-glutamyl transpeptidase (GGTP) activity, and the clinical setting in a group of patients with elevated total ALP activity. In addition, expected ranges of serum ALP isoenzymes for healthy young men and also for a geriatric population are presented.     

Clinical assessment of the electrophoretic separation of alkaline phosphatase isoenzymes

Over a five-month period, using data from patients in whom alkaline phosphatase (ALP) isoenzyme studies were requested routinely, we compared actual clinical diagnoses with the predicted diagnoses based on the results of electrophoretic separation of ALP isoenzymes on cellulose acetate before and after heat treatment and on elevated enzymatic activity of gamma glutamyl transferase (GGT) and alanine aminotransferase (ALT) activity. ALP isoenzymes were interpreted on a qualitative basis (presence or absence of liver, bone, or other isoenzyme) by individual clinical pathologists. Overall, the consistency of agreement in 61 patients was 66% for GGT, 51% for ALP isoenzymes, and 21% for ALT. In 44 patients with definite diagnoses, the sensitivity and specificity, respectively, of each laboratory test for patients with liver disease were 88 +/- 5.7% and 64 +/- 14.5% (ALP isoenzymes); 88 +/- 5.7% and 91 +/- 8.6% (GGT); and 6 +/- 4.1% and 91 +/- 8.6% (ALT). In patients with bone disease, the sensitivity and specificity of ALP isoenzymes was 75 +/- 10.8% and 86 +/- 6.6%, respectively. The results indicate that isoenzymes as currently performed need to be improved through standardization of the interpretation of ALP isoenzyme patterns to establish uniformity of comments.     

Humoral and cellular immune mechanisms of neutropenia in patients with Felty's syndrome.

The objective of these studies was to assess the relative role of humoral or cellular immune mechanisms in the neutropenia of Feltry's syndrome. The amounts of neutrophil-bound IgG were measured by a sensitive antiglobulin inhibition assay. These values were increased in eight out of seventeen patients with Felty's syndrome over the highest values seen with seventeen patients with rheumatoid arthritis without leukopenia or twenty-six normal individuals. The levels of serum IgG neutrophil-binding activity from thirteen out of twenty-one patients with Felty's syndrome were greater than the highest values seen with normal sera compared to six out of thirty sera from patients with rheumatoid arthritis alone. A strong correlation was observed between the levels of direct neutrophil-bound IgG and the serum IgG neutrophil-binding activity for the seventeen patients with Felty's syndrome. Inhibition of normal marrow granulocyte colony growth greater than 40% was seen with the peripheral blood mononuclear leucocytes from five out of eleven patients with Feltry's syndrome. All five patients had normal levels of neutrophil bound IgG and normal to moderately increased levels of serum IgG neutrophil binding activity. In contrast, of the six patients whose mononuclear leucocytes did not inhibit granulocyte colony growth, three had markedly elevated levels of neutrophil-bound IgG and four had elevated levels of serum IgG neutrophil binding activity. These results suggest that humoral and cellular immune mechanisms of neutropenia may be present in different subsets of patients with Felty's syndrome.     

Use of alkaline phosphatase isoenzyme analysis in the evaluation of cholestatic liver disease

A useful laboratory test for the differentiation of liver, bone, and intestinal alkaline phosphatase (ALP) isoenzymes in serum is presented. Electrophoresis in polyacrylamide gel is performed with untreated serum as well as with serum incubated at 56 degrees C for 10 min. The heating step denatures bone isoenzyme which may obscure the liver ALP band when present in large amounts. Visualization of ALP activity is accomplished by the use of buffered p-toluidinium 5-bromo-4-chloro-indolyl phosphate and magnesium ions. In serum of patients with cholestatic liver disease, the occurrence of large molecular weight liver cell membrane fragments which contain ALP activity is postulated. These ALP-containing fragments occur at the origin of the electrophoretogram, unable to penetrate the small pore separation gel. Abnormalities involving ALP isoenzymes, such as bone isoenzyme arising from increased osteoblastic activity, may be detected. Intestinal isoenzyme, normally present in small amounts in some subjects of blood groups B or O, may be elevated in certain liver diseases, such as cirrhosis. By the use of this method the routine question of whether an ALP found to be increased in a screening procedure is due to liver or bone abnormality may be answered. In addition, the occurrence of abnormal ALP bands arising from cholestatic conditions and the occurrence of abnormal amounts of intestinal isoenzyme may also be detected.     

Adenosine deaminase isoenzyme levels in patients with human T-cell lymphotropic virus type 1 and human immunodeficiency virus type 1 infections.

In serum, the enzyme adenosine deaminase (ADA) is known to be divided into two isoenzymes, ADA1 and ADA2, which have different molecular weights and kinetic properties. The present study investigated ADA isoenzyme levels in the sera of patients infected with retroviruses associated with adult T-cell leukemia (ATL), human T-cell lymphotropic virus type 1 (HTLV-1)-associated myelopathy (HAM), and AIDS, ADA isoenzyme activities were found to be significantly (P < 0.001) higher in the sera of patients with ATL, HAM, and AIDS than in the sera of healthy controls. In the case of the ADA subtypes in the sera of patients with ATL, ADA1 activity was significantly (P < 0.001) elevated in patients with the acute and lymphoma types of ATL compared with that in patients with the chronic and smoldering types of ATL. ADA2 activity was significantly elevated in the sera of patients with the acute, lymphoma, and chronic types of ATL (P < 0.001) compared with that in patients with smoldering ATL and HTLV-1 carriers. In the case of patients with human immunodeficiency virus type 1 (HIV-1) infection, ADA1 and ADA2 activities in the sera of patients with AIDS and HIV-1 antibody-positive individuals were significantly (P < 0.001) higher than those in the sera of HIV-1 antibody-negative individuals. A significant elevation in ADA2 activity was also seen in the sera of AIDS patients (P < 0.01) compared with that in the sera of HIV-1 antibody-positive individuals. These results suggest that the magnitude of elevation of ADA isoenzyme levels in serum correlates well with the clinical conditions of the patients with these diseases. Measurement of the activities of ADA isoenzymes may therefore provide an additional parameter for distinguishing the subtypes of ATL and may prove to be useful as prognostic and therapeutic monitors in diseases associated with HTLV-1 and HIV-1 infections.     

Serum interferon in Navajo children with severe combined immunodeficiency disease inhibits lymphoblastogenesis

Two Navajo Indian children with severe combined immunodeficiency disease (SCID) lost reconstituted immune function after virus infections. A serum factor which inhibited normal lymphocyte response to mitogens was found in one of them and led to the examination of sera from five other Navajos with SCID. Mean inhibition by six Navajo sera was 67%; no inhibitor was found in sera from normal adults and children. The inhibitor activity was nondialyzable and heat stable, yet partially sensitive to pH 2.0, suggesting that interferon(s) was present. Interferon (IFN) activity in patient sera ranged from 10 to 300 U/ml. Normal children had peak serum IFN levels of 100 and 30 U/ml in the acute and convalescent periods, respectively, of virus infections. IFN alpha, IFN beta, and IFN gamma were identified in SCID sera by specific antisera. Both inhibitor and IFN activities in three Navajo sera were 88-95 and 89-100%, respectively, removed with anti-IFN antisera. Similar patterns of inhibition of lymphoblastogenesis were seen with IFN standards. IFN levels in the SCID patients did not correlate with documented infections; elevated levels were present when no infections could be documented. The immunologic imbalances in some forms of SCID may be related to circulating inhibitors, possibly interferon.     

A study of serum immunoglobulins including IgE and reaginic antibodies in patients with bronchial asthma in relation to the spectrum of the disease

Immunoglobulins G, A, M, D, and E were studied in the sera obtained from sixty-nine bronchial asthma patients, who were graded objectively according to atopic score. Among other associated atopic diseases, they often had allergic rhinitis. Ninety-one per cent of these patients had elevated serum IgE levels and the mean level of serum IgE was more than 3.5 times higher than that observed in the normal subjects. Moreover, as their atopic score increased, the serum IgE levels were also elevated, and every patient with high atopic score, had elevated serum IgE levels: This indicates the association of atopic state with serum IgE level. Furthermore, reaginic antibodies were studied in some of these patients by intradermal tests, Prausnitz-Küstner (PK) reaction, conjunctival and bronchial provocation tests. Intradermal and PK tests were comparable and proved to be the most sensitive method of detecting reagins. The intensity of these reactions correlated significantly with serum IgE level, although this relationship is not invariably present. The reaginic activities in the sera of the patients with atopic bronchial asthma also compared well with the positive bronchial test (induced asthma) by inhalation of specific aerosol although this association is also not always present. Eleven patients with high atopic score had isolated serum IgA deficiency. There was a significant rise of mean serum IgM in comparison to the controls.     

The relationship between anthropometry and serum concentrations of alkaline phosphatase isoenzymes, liver-enzymes, albumin, and bilirubin

Alkaline phosphatase (ALP) is present in human preadipocytes. The aim of this study was to determine the relationship between anthropometry and serum levels of ALP isoenzymes, liver enzymes, albumin, and bilirubin. Anthropometric variables; serum total, bone, liver, and intestinal ALP levels; and alanine aminotransferase (ALT), albumin, total protein, total bilirubin, and g-glutamyltransferase serum levels were measured in 100 volunteers. The levels (given as median [interquartile range]) for total (74.0 U/L [30.0 U/L] vs 62.0 U/L [22.0 U/L]; P <.05) and liver ALP (37.3 U/L [14.6 U/L] vs 26.1 U/L [12.0 U/L]; P < .05) were higher in obese than in lean subjects. The levels of the other ALP isoenzymes and blood analytes were not significantly different between these groups. Albumin and ALT were the only blood proteins studied with serum levels that correlated significantly with waist circumference. This present study demonstrates a relationship between abdominal obesity and serum ALT levels and between body mass index and ALP levels. These findings suggest that serum ALP, particularly liver ALP, is derived from adipose and hepatic tissue.     

Isoenzymes of alkaline phosphatase in epileptic patients receiving carbamazepine monotherapy.

The effects of carbamazepine monotherapy were investigated in 20 female and 21 male epileptic patients to determine whether treatment would induce an increase in serum alkaline phosphatase (ALP) activity, a known effect of many anticonvulsant drugs. Serum total ALP activity was increased in nine out of the 41 patients (22%), serum bone ALP activity was increased in 10 (24%), and serum non-bone ALP activity was increased in three (7%). There was no significant difference when the mean of the patients' serum total ALP was compared with that of the controls. Twenty per cent of the patients with increased serum bone ALP had normal serum total ALP, indicating that increased serum bone isoenzyme activity may precede an increase in the total enzyme activity. This should be considered when interpreting results of increased total ALP in epileptic patients.     

The role of food allergens in childhood asthma

Eighty-eight patients' sera with allergen-specific IgE levels elevated only to food allergens were collected between October 1997 and March 2002 at the National Taiwan University Hospital. Thirty-three of the patients fulfilled the diagnostic criteria of asthma and were included. Most (72.7%) patients had elevated serum allergen-specific IgE levels only to one food allergen. The most common food allergens were milk and egg white. The patients with elevated soy bean-specific IgE levels had significantly higher levels of serum food allergen-specific IgE than those with either elevated milk or egg white-specific IgE levels. This study investigated some food allergen responses of asthmatic patients whose serum allergen-specific IgE levels were elevated only to food allergens. The results suggested that the allergic asthmatic response in our patients was most likely related to food rather than aeroallergens or fungal allergens.     

Expression of the major rheumatoid factor cross-reactive idiotype in pediatric patients with systemic lupus erythematosus

Rheumatoid factor cross-reactive idiotype (RF-CRI) is expressed in high concentrations in the sera of some patients with rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). To determine if RF-CRI is specifically expressed in rheumatic disease or if it is secondary to polyclonal B-cell activation, we examined sera of 23 children with SLE, 16 adolescents with infectious mononucleosis (IM), and age-matched pediatric controls for RF-CRI expression. Concentrations of RF-CRI in serum, determined by an inhibition ELISA, were 24 +/- 17 micrograms/ml (mean +/- SD) in 25 normal children, 31 +/- 17 in 16 young adults with IM, and were significantly increased, 70 +/- 80 micrograms/ml, in the 23 children with SLE (p less than 0.036). Eleven of 23 SLE patients had serum RF-CRI greater than the mean +/- 2 SD for normal children. Ten of 23 SLE sera contained IgM rheumatoid factor (RF) activity. One patient with IM had a borderline elevated RF-CRI level, and 5 IM patients had RF in their sera. The serum IgM concentrations in sera were: SLE (192 +/- 93 mg/dl) and IM (234 +/- 77 mg/dl) sera. These levels were significantly elevated compared to controls (132 +/- 44 mg/dl), p less than 0.031 for SLE and p less than 0.001 for IM, suggesting that polyclonal activation of B cells was present in SLE and IM patient groups. Increased expression of RF-CRI in the SLE patients correlated directly with high titer anti-DNA antibody values (r = 0.3965, p less than 0.05) and RF activity when human IgG (r = 0.5026, p less than 0.05) was used as the RF binding substrate and inversely with serum C3 levels (r = 0.3925, p less than 0.05). RF-CRI expression did not correlate with RF that bound rabbit (r = 0.3123, p greater than 0.05). Increased serum RF-CRI expression is not a result of polyclonal B-cell activation. RF-CRI may be selectively up-regulated in patients with SLE.     

孕妇血清中钙、磷、碱性磷酸酶与全血骨碱性磷酸酶检测结果分析

目的通过检测孕妇血清中钙（Ca）、磷（P）、碱性磷酸酶（ALP）、全血骨碱性磷酸酶（BALP）,早期预防婴幼儿佝偻病的发生。方法对2005年12月－2007年10月来我院就诊的孕早、中、晚期的孕妇采血进行Ca、P、ALP、BALP的测定。结果与结论孕妇血清中Ca、P、ALP、BALP血清水平在不同孕期存在差异。     

Prevalence and persistence of C1q binding activity in healthy subjects.

Samples of serum from 885 normal healthy blood donors were tested for the presence of soluble immune complex-like material by a solid-phase C1q binding assay. The majority of donors (93%) had low or undetectable levels of C1q binding activity in their sera, but 6% had levels that were clearly outside the normal distribution. When these individuals were retested after several weeks half of them still had elevated levels of C1q binding activity.     

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with colorectal cancer

Various isoenzymes of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) exist in human colorectal mucosa. In our last experiments we have shown that ADH and ALDH are present also in colorectal cancer cells. Moreover the activities of total ADH and class I isoenzymes were significantly higher in cancer tissue than healthy mucosa. This may suggest that these changes may be reflected by enzyme activity in the serum. Therefore, we have measured the activity of total ADH, and classes I-IV of this enzyme and ALDH in the sera of patients suffering from this cancer. Total ADH activity was measured by a photometric method with p-nitrosodimethylaniline (NDMA) as a substrate and ALDH activity by the fluorometric method with 6-methoxy-2-naphtaldehyde as a substrate. For the measurement of the activity of class I and II isoenzymes we employed fluorometric methods, with class-specific fluorogenic substrates. The activity of class III ADH was measured by the photometric method with formaldehyde and class IV with m-nitrobenzaldehyde as a substrate. Serum samples were taken for routine biochemical investigations from 52 patients with colorectal carcinoma before treatment. A statistically significant increase of class I ADH isoenzymes was found. Therefore the total ADH activity was also significantly increased. The total ALDH and the activity of other tested ADH isoenzymes were unchanged. We also observed the increasing tendency of ADH I activity in accordance with the advance of disease. The activity of class I ADH isoenzymes was elevated in the serum of patients with colorectal cancer. This activity was derived from colorectal cancer cells and probably from severely damaged liver by metastatic disease.     

去势大鼠5种骨转换生化指标的动态变化

目的： 对去势大鼠骨质疏松动物模型形成过程中5种骨转换指标的变化及其相关性进行研究。方法: 将3月龄SD雌性大鼠分为3组：切除卵巢组(OVX)，假手术组(sham)和对照组(control)，术前和术后分别于1、1.5、2、2.5、3和4月检测血清骨钙素(OC)、碱性磷酸酶(ALP)、 骨碱性磷酸酶(BALP)、抗酒石酸酸性磷酸酶(TRAP)和羟脯氨酸(HYP)水平，并作大鼠胫骨病理切片检查。结果: OVX组血清OC、ALP、BALP、TRAP和HYP水平均明显高于sham组，其变化顺序依次为：TRAP/HYP→OC→ALP/BALP；5种指标之间呈显著正相关；术后3月OVX组大鼠胫骨小梁结构有病理改变。结论: 去势大鼠属于高转换型骨质疏松；在模型形成过程中，骨吸收指标的变化早于骨形成指标的改变；骨转换指标是反映绝经后早期骨量丢失的灵敏指标。     

Bone-specific alkaline phosphatase as a good indicator of bone formation in sheepdogs

Eight clinically healthy male sheepdogs were selected and subjected to experimental radius transaction. Blood samples were taken before and weekly after surgery, and radiographs were taken immediately and weekly after surgery to trace new bone formation. Total protein, total ALP, and bone-specific ALP were measured in serum samples. There was no significant difference in total proteins, but total ALP and bone ALP activity were significantly increased (P < 0.05) after surgery (during bone formation).We found a weak correlation between total ALP activity and BALP activity during the study. The bridging callus was completed in the fourth week of the experiment, and the gap was fully filled with new bone. However, increased levels of total ALP and bone-specific ALP activity were maintained throughout the study and did not reduce at 4 weeks. We concluded that serum activity of bone-specific ALP is a good indicator of bone formation in sheepdogs, and when radiographs inform the completion of bone fracture healing, cellular activity in the healing region is likely to be continuing.     

Gaucher disease: comparative study of acid phosphatase and glucocerebrosidase in normal and type-1 Gaucher tissues

Acid phosphatase activity was determined in serum, cultured fibroblasts, and peripheral blood lymphocytes of six splenectomized adult patients with non-neuropathic Gaucher disease in two Canadian families. Elevated levels of serum acid phosphatase activity (520-711% of normal) were found in four patients who also developed orthopedic complications associated with Gaucher disease, including intermittent bone pain, arthritis, collapse of femoral head, and pathological fractures. Serum acid phosphatase activity in two patients who do not have bone involvement were found to be within the normal range. Contrary to the serum enzyme, acid phosphatase activity in lymphocytes and cultured fibroblasts of all of the patients was within the normal range. Deficient glucocerebrosidase (7.5-15.5% of normal) and acid beta-glucosidase (13.8-27.8% of normal) activities were noted in all probands. Similarly, normal levels of fibroblast and lymphocyte acid phosphatase activity were found in Gaucher heterozygotes whose glucocerebrosidase activity was about 50% of normal. Acid polyacrylamide gel electrophoresis and acid phosphatase activity staining of the patients' sera showed that the elevated acid phosphatase is isozyme type 5 osteoclastic origin. The apparent Michaelis constant, Km, of fibroblast glucocerebrosidase for the natural substrate was 0.6 +/- 0.1 mM for controls and 0.6 +/- 0.2 mM for the patients. These data suggest that the assay of serum acid phosphatase activity for the presumptive diagnosis of Gaucher disease is not completely reliable and that the elevated level of serum acid phosphatase in Gaucher disease is most likely a secondary phenomenon which may be indicative of bone involvement in some patients with this disorder. It also demonstrates the clinical heterogeneity of type 1 Gaucher disease, even among full sibs of the same heterozygous parents.     

Bone and cartilage turnover markers, bone mineral density, and radiographic damage in men with ankylosing spondylitis

PURPOSE: To determine the levels of bone and cartilage turnover markers in men with ankylosing spondylitis (AS) and to investigate their associations with disease activity, bone mineral density, and radiographic damage of the spine. PATIENTS AND METHODS: This cross-sectional study enrolled 35 men with newly diagnosed AS. The bone mineral densities (BMD) of their lumbar spines and proximal femurs, Bath AS Disease Activity Index (BASDAI), and Bath AS Radiographic Index (BASRI) were evaluated. Urinary C-terminal telopeptide fragments of type I collagen (CTX-I) and type II collagen (CTX-II) levels were determined by enzyme-linked immunosorbent assay, and serum levels of bone-specific alkaline phosphatase (BALP) and osteocalcin were determined by an enzyme immunoassay. Levels of biochemical markers were compared with those of 70 age-matched healthy men. RESULTS: Patients with AS had significantly higher mean urinary CTX-I and CTX-II levels than control subjects (p<0.05). Elevated urinary CTX-I levels correlated well with BASDAI, femoral BMD, and femoral T score (p<0.05), and elevated urinary CTX-II levels correlated well with spinal BASRI (p<0.05) in patients with AS. Mean serum BALP and osteocalcin levels did not differ between patients and controls and did not show any significant correlations with BMD, BASDAI, or BASRI in men with AS. CONCLUSIONS: Elevated CTX-I reflects disease activity and loss of femoral BMD while elevated CTX-II levels correlate well with radiographic damage of the spine, suggesting the usefulness of these markers for monitoring disease activity, loss of BMD, and radiographic damage in men with AS.     

Séparation électrophorétique des isoenzymes de la phosphatase alcaline par le Kit Hydragel 15 Iso-PAL®

We evaluated a new electrophoresis Kit (Hydragel 15 Iso-PAL®, Sebia) for the separation of alkaline phosphatase (ALP) isoforms. This agarose gel technique uses wheat germ lectin affinity for sialic acids present on ALP isoenzymes. Similar results were obtained between serum and plasma (lithium heparinate), particularly for the bone fraction (< 5% difference). In healthy subjects, bone and liver H1 fractions are the two major components of ALP activity (up to 90%) with a ratio close to 1/1. This technique displays a practical advantage for laboratories, due to the absence of sample pre-treatment, the use of a single gel and automation on the Hydrasys® system (2 h analysis time for 7 patients). This method also offers a clinical advantage for bone and hepatobiliary diseases, allowing a whole panorama of ALP activity.