Amyloid Substance Research Articles

Hepatic amyloidosis in intravenous drug abusers and AIDS patients

The clinical and pathological features of four liver biopsies and 12 autopsies from 1981-1990 with hepatic amyloidosis were reviewed. All of the patients had a history of both intravenous and subcutaneous cocaine and heroin use with chronic suppurative skin ulcers. Five patients were proven to have the acquired immunodeficiency syndrome at autopsy. Systemic amyloidosis was diagnosed in only five out of the 16 patients prior to death. Hepatomegaly was present in 12 patients. The amyloid protein was AA in 14 and AL in one case. Definitive characterization of the amyloid substance was not possible in one case. There was no evidence of multiple myeloma or a plasma cell dyscrasia in the one patient with AL amyloid. The amyloid distribution in the liver was both parenchymal and vascular in 12 cases. Cholestasis, which appeared to occur preterminally, was the only histological finding that correlated with the patient's clinical condition. The histological pattern of amyloid in the liver could not predict the type of amyloid protein that was identified. Amyloidosis should be considered in the differential diagnosis of unexplained hepatomegaly in the acquired immunodeficiency syndrome with chronic suppurative infections.

Primary Localized Amyloidosis in One Family

Primary localized amyloidosis was found in a family. A 66-year-old woman had suffered from hoarseness for 18 years. A biopsy specimen from the larynx showed amyloid deposits in the submucosal connective tissue. Her 40-year-old daughter noticed a subcutaneous nodule on her philtrum. Histologic examination showed the deposition of amyloid in the middle and lower dermis, which also encased the blood vessels and epidermal appendages. In both cases the amyloid deposits were positively stained with Congo red and the staining was resistant to potassium permanganate treatment. These amyloid deposits were strongly positive against the anti-amyloid antibody of the lambda light chain. These results indicate that the amyloid substance is derived from protein AL. There was no clinical or laboratory evidence of systemic amyloidosis or multiple myeloma in either patient.

Amiloidose traqueo-brônquica difusa.

The case of a 52 year old man, whose initial clinical manifestations were dyspnea, bloodstained sputum and malaise is reported. After the initial cancer hypothesis, a diagnosis of diffuse primary tracheo-bronchial amyloidosis was made. The amyloid substance present was not of A A type and the plasma cells next to the deposits were polyclonal. The piece-meal removal of the masses by bronchoscopy led to profuse bleeding. The patient died with sepsis. The clinical, pathological and therapeutical aspects of lower respiratory tract amyloidosis are reviewed.

Islet Amyloid Polypeptide has No Effect on Amylase Release from Rat Pancreatic Acini Stimulated by CCK-8, Secretin, Carbachol and VIP.

Islet amyloid polypeptide (IAPP), a novel peptide recently isolated from the amyloid substance of pancreatic islets, has been localized in normal islet B cell granules, suggesting its physiological action in exocrine pancreatic function. In the present study, the direct effect of IAPP on secretagogue-stimulated pancreatic enzyme secretion was investigated using dispersed rat pancreatic acini. Various concentrations of IAPP (10(-5) -10(-10) M) showed no significant influence on amylase release stimulated either by submaximal doses of CCK-8 (10(-11) M), secretin (0.5 x 10(-6) M) or carbachol (10(-6) M), or by a maximal dose of vasoactive intestinal peptide (10(-8) M). These results may provide negative evidence for a physiological significance of IAPP in rat pancreatic enzyme secretion.

Keratin in cutaneous amyloidoses

When Congo red stain was the only criterion, “amyloid” was considered to be a single substance. With the advent of electron microscopy, this idea was further strengthened because unfortunately the ultrastructure of all amyloids including those of the skin was found to be uniform. Clinical dermatologists knew that there were two types of skin amyloidoses; one manifesting itself in the skin as well as in various internal organs, the other being exclusively limited to the skin. The first group included amyloidoses associated with multiple myeloma (now termed AL amyloid), infections (now termed AA amyloid), and familial hereditary disease (of chemically variable composition). The second group consisted of truly cutaneous diseases such as lichen amyloidosus, macular amyloidosis, and cutaneous epithelioma-associated amyloidoses. Nodular amyloidosis was thought to be the link between these two groups because occasionally it can become systemic disease; it turned out to be a systemic disease of immunoglobulin light-chain production with a limited skin manifestation. Today we realize that this clinical division was more important than histologic and ultrastructural analyses of amyloid deposits: the second group of amyloidoses occur exclusively in the skin because the amyloid substances come from a special degeneration of keratinocytes of the epidermis and its derivatives.

The amino terminal portion of cerebrospinal fluid cystatin C in hereditary cystatin C amyloid angiopathy is not truncated: direct sequence analysis from agarose gel electropherograms

The isolated amyloid substance in hereditary cystatin C amyloid angiopathy (HCCAA) is mainly composed of a cystatin C variant devoid of the 10 amino terminal amino acid residues of extracellular cystatin C from healthy individuals. We have developed a procedure for protein sequencing directly from agarose gel electropherograms and used this in conjunction with isoelectric focusing to investigate the amino terminal sequence of cerebrospinal fluid (CSF) cystatin C in HCCAA patients. The amino-terminal sequence determined for cystatin C from a HCCAA patient CSF sample, Xaa-Ser-Pro-Gly-Lys-Pro-Pro-Xaa-Leu-Val-Gly-Gly-Pro-Met-Xaa-Ala-Xaa-Val, showed that the protein was not amino-terminally truncated. CSF cystatin C from all nine HCCAA patients investigated was found to have an isoelectric point identical to that of native cystatin C, and the truncated form of cystatin C isolated from amyloid deposits was shown to contribute to less than 1% of the total amount of cystatin C in CSF. The total cysteine proteinase inhibitory capacity of CSF from HCCAA patients was lower than that of CSF from other patients. This decreased CSF inhibitory capacity in HCCAA patients was caused by decreased levels of cystatin C, since the levels of the other two cysteine proteinase inhibitors found in CSF, alpha 2-macroglobulin and kininogen, were significantly higher than in CSF from non-HCCAA patients.

気道アミロイドーシスの免疫組織学的検討

Although amyloid proteins show an identical figure of a β-pleated sheet electro-microscopically, it has been demonstrated that these proteins originate from different precursor proteins. Recently, they have been classified on the basis of their biochemical and immunochemical characteristics and their precursor proteins.We examined two patients with respiratory amyloidosis (a 57-year old female and a 64-year old male). Their chief complaints were gradually progressing hoarseness and dysphonia. Amyloid deposits were found as yellow plaques on the mucosa of the respiratory tract extending from the epipharynx to the bronchi. They were removed endoscopically and vaporized with a CO2-laser.The origin of the amyloid protein was investigated histochemically and immunochemically. The Amyloid substances in both patients were derived from the A-chain of light chain immunoglobulins.

舌に腫りゅうの形成を認めた原発性アミロイドーシスの１症例

Amyloidosis is a lesion in which an amyloid substance deposits in various organs and tissues of the body.Recently, we experienced a case of primary amyloidosis forming multiple nodules in the tongue. The patient was a 74-year-old male.Multiple nodules, 5-10mm in diameter, were observed in the tongue and the lingual frenulum. Histopatliologically, the excised nodules were diagnosed as amyloidosis. The amyloid protein was identified as protein AL by potassium permanganate treatment.

The amino terminal portion of cerebrospinal fluid cystatin C in hereditary cystatin C amyloid angiopathy is not truncated: direct sequence analysis from agarose gel electropherograms

The isolated amyloid substance in hereditary cystatin C amyloid angiopathy (HCCAA) is mainly composed of a cystatin C variant devoid of the 10 amino terminal amino acid residues of extracellular cystatin C from healthy individuals. We have developed a procedure for protein sequencing directly from agarose gel electropherograms and used this in conjunction with isoelectric focusing to investigate the amino terminal sequence of cerebrospinal fluid (CSF) cystatin C in HCCAA patients. The amino-terminal sequence determined for cystatin C from a HCCAA patient CSF sample, Xaa-Ser-Pro-Gly-Lys-Pro-Pro-Xaa-Leu-Val-Gly-Gly-Pro-Met-Xaa-Ala-Xaa-Val, showed that the protein was not amino-terminally truncated. CSF cystatin C from all nine HCCAA patients investigated was found to have an isoelectric point identical to that of native cystatin C, and the truncated form of cystatin C isolated from amyloid deposits was shown to contribute to less than 1% of the total amount of cystatin C in CSF. The total cysteine proteinase inhibitory capacity of CSF from HCCAA patients was lower than that of CSF from other patients. This decreased CSF inhibitory capacity in HCCAA patients was caused by decreased levels of cystatin C, since the levels of the other two cysteine proteinase inhibitors found in CSF, alpha 2-macroglobulin and kininogen, were significantly higher than in CSF from non-HCCAA patients.

What role(s) may extracellular matrix, particularly heparan sulfate, play in amyloid of Alzheimer's disease

The authors of this article present and attempt to substantiate the thesis that proteoglycan(s), mainly heparan sulfate, are an important ingredient in the pathogenesis of the amyloid found in persons with Alzheimer's disease. Evidence presented indicates that glycosaminoglycans are regular constituents of many amyloid substances including that of senile plaques and congophilic angiopathy of Alzheimer's disease. It is proposed that the proteoglycans play a role in amyloidogenesis by one or a combination of the following mechanisms: 1) inducing amyloid fibrils containing a predominant beta-pleated sheet structure, 2) influencing amyloid deposition to occur at specific tissue sites and/or 3) prevent amyloid degradation.

Dystrophic peptidergic neurites in senile plaques of Alzheimer's disease hippocampus precede formation of paired helical filaments

The relationship between peptidergic dystrophic neurites and paired helical filament (PHF)-positive neurites in Alzheimer's disease (AD) senile plaques (SPs) was studied using combined fluorescence and bright-field optics. Cryostat sections of AD hippocampi were first stained with thioflavine-S and immunolabelled with antisera raised against different neuropeptides: somatostatin-28(1–12), somatostatin-14, neuropeptide Y, cholecystokinin (CCK) and substance P. Secondly, using the elution-restaining procedure, sections were immunolabelled with anti-tau/PHF. In immature SPs, clusters of abnormal, swollen neurites were found. The dystrophic, strongly peptidic-positive neurites contained fewer PHFs than the poorly positive ones. Cell bodies, exhibiting a peptidic content, could be found within SPs without any alteration. These results suggest the following sequence of events: an extracellular poisoning mechanism, perhaps the amyloid substance, first changes the structure of presynaptic endings and causes the formation of ballooning dystrophic neurites filled with their normal peptidic content. Subsequently, intracellular degradation occurs with formation of the PHFs. Then the other structures such as dendrites and perikarya are damaged by the same mechanism. Therefore, this phenomenon seems to precede any formation of PHFs in SPs.

Senile plaque-like structures: observation of a probably unknown type of senile plaque by periodic-acid methenamine silver (PAM) electron microscopy

Numerous diffuse senile plaque-like structures (SPLSs) were found in the cerebral cortex from cases with dementia of the Alzheimer type by means of the methenamine-Bodian method. SPLSs varied in shape and size. They were never recognized in the original Bodian, PAS and Congo red preparations, but were positive with anti-beta-protein immunostaining and periodic-acid methenamine silver (PAM) methods, which are thought to specifically stain amyloid substance. With PAM electron microscopy, we found sparse aggregations of amorphous, often ramified, structures with fine granular silver deposits in SPLS. Routine electron microscopic examination on the same portion where SPLS were confirmed by PAM electron microscopy revealed amorphous, partially fibrous structures. These structures might be amyloid or amyloid-precursor substance. In SPLSs only a few degenerated neurites and astrocytic processes with glycogen granules were seen. We consider SPLSs to be a kind of senile plaque.

Age-associated changes in renal glomeruli of mice

To investigate age-associated changes in renal glomeruli of C57BL/6 female mice, we used a single radial immunodiffusion method to measure albumin excretion. Up to 100 mg/dl in urine samples was regarded as microalbuminuria. The mean amount of urinary albumin increased from 14.0 mg/dl at 6 months to 151.1 mg/dl at 24 months of age. Microalbuminuria occurred in 64.6% of tested mice by the time they were 24 months old, and 10% of the mice had marked albuminuria (more than 100 mg/dl) at that time. Parallel morphological study showed that renal mesangial changes were also age-dependent. Mesangial cell proliferation and spike lesions in glomerular capillary walls appeared in aged mice with microalbuminuria, and were then followed by diffuse glomerular sclerosis accompanied by marked albuminuria. Histological scores on damage in the renal mesangium with changes of glomerular basement membrane increased significantly with age from a mean score of 0 at 6 months to 3.24 at 24 months of age. Immunofluorescent study showed a marked deposition of IgG and IgM, but no complement component C3 in enlarged mesangium. Electron microscopic examination of diffuse sclerotic glomeruli in aged mice revealed amyloid substances. These results suggest that assays of albuminuria could be a useful method for early detection of age-associated renal deterioration.

Hemorrhagic Bullous Amyloidosis

• In patients with bullous hemorrhagic amyloidosis of the skin, the skin lesions were the first manifestations of a plasma cell dyscrasia. Both cases were characterized by similar clinical, histologic, and ultrastructural findings showing an intradermal blister within deposits of amyloid substances. Immunohistologic investigations with a panel of antibodies directed against amyloid fibril proteins showed reactivity of the amyloid deposits with an anti-A X serum in both patients. (<i>Arch Dermatol</i>1988;124:1683-1686)

Hemorrhagic bullous amyloidosis. A histologic, immunocytochemical, and ultrastructural study of two patients

In patients with bullous hemorrhagic amyloidosis of the skin, the skin lesions were the first manifestations of a plasma cell dyscrasia. Both cases were characterized by similar clinical, histologic, and ultrastructural findings showing an intradermal blister within deposits of amyloid substances. Immunohistologic investigations with a panel of antibodies directed against amyloid fibril proteins showed reactivity of the amyloid deposits with an anti-A lambda serum in both patients.

The clinical and biochemical spectrum of hereditary amyloidosis

Familial amyloidosis, once described as a puzzling and highly unusual form of polyneuropathy, is now recognized to be a collection of familial diseases with usually autosomal-dominant inheritance and widespread ethnic distribution. Familial amyloidosis occurs throughout the world and encompasses an extremely broad spectrum of clinical manifestations. In some families, progressive peripheral neuropathy dominates the illness, while in others, renal failure, ocular amyloid deposits, cardiac decompensation, or intracranial hemorrhage is the most significant clinical feature. The Portuguese (type I) and the Iowa (type III) neuropathies characteristically begin with lower limb involvement, while in the Indiana (type II) form, upper limb neuropathy is seen first; in the Japanese families with familial amyloid polyneuropathy, symptoms first become evident around age 40, whereas in the Texas family, onset is in the seventh decade. The prognosis for the different families is highly variable. Current classification of the familial amyloid polyneuropathy syndromes is based on their characteristic clinical presentations, but ongoing biochemical identification of the protein composition of amyloid substance in each form will make a more rational nosology feasible in the near future. To date, no therapy has been shown to arrest or reverse the progressive accumulation of amyloid deposits in most forms of familial amyloidosis. Familial Mediterranean fever is a major exception, and the incidence of amyloidosis associated with this disease has been dramatically reduced by the widespread prophylactic use of colchicine. Technology currently available permits the reliable identification of asymptomatic relatives at risk for developing amyloid neuropathy as well as the prenatal identification of carriers of the mutant transthyretin gene. These strategies can be used in genetic counseling aimed at reducing the continued propagation of the mutant gene.

臼後三角部，舌，頬粘膜に認められたアミロイドーシスの１例

A case of primary amyloidosis diagnosed by morphological and immunological examination was reported. The patient was a 71 year-old woman who had tumorous masses at the retromolar pad, the left buccal mucosa and the tongue. The specimens taken from the lesions displayed green birefriegence after Congo red staining on polarization microscopy, and proliferation of plasma cell around the amyloid substance was obsereved. Also, positive reaction of λ type antisera with those cells, as well as the amyloid substance were shown by immunohistochemical method. Besides, the quantitative analysis of immunogloblin and immunoelectrophoresis of the patient's serum wre examined; increased IgGλ type monoclonal protein was revealed. However, there were no findings associatedwith multiple myeloma by several systemic examinations, e. g., X-ray and Tc-scintigram. From those findings described above, this case was finally diagnosed as “primary amyloidosis with plasma cell dyscrasia”.

Cortical angiopathy in Alzheimer's disease: the formation of dystrophic perivascular neurites is related to the exudation of amyloid fibrils from the pathological vessels.

We studied the organization of dystrophic neurites around pathological vessels in Alzheimer cortex. Two techniques were used simultaneously on serial sections: thioflavine staining of amyloid substance and immunohistochemistry with immune sera against Paired Helical Filaments (anti-PHF) and native Tau proteins (anti-Tau). We observed different distributions of dystrophic neurites (immunolabelled with anti-PHF or anti-Tau) around thioflavine-stained angiopathic arterioles. The wall of the vessels with large diameter (greater than 100 microns) presented a congophilic angiopathy without neuropil reaction. In vessels with lesser diameter (less than 100 microns), dystrophic neurites constituted a discontinuous sleeve around vessels, always in close contact with amyloid substance outside the wall (dysphoric angiopathy). We observed structures similar to senile plaques around capillaries (diameter: 10-15 microns). The sleeve of dystrophic neurites with aggregated Tau proteins were always observed in the close vicinity of the amyloid substance which exuded from the pathological blood vessels. Thus, the exudation of these amyloid fibrils seems to induce the formation of dystrophic neurites (neuritic reaction).

Transformation of degenerating neurofibrils into amyloid substance in Alzheimer's disease: Histochemical and immunohistochemical studies

Degenerating neurofibrils (DNF), which are composed of paired helical filaments (PHF) and amyloid fibrils (AF), are the 2 characteristic pathological fibrillar deposits in Alzheimer cortex. These fibrils were simultaneously studied by 2 techniques: The immunolabelling with a specific antiserum raised against PHF and elective thioflavine S staining of AF. In neuronal perikaryons, neurofibrillary tangles (NFT) consist of 3 populations: firstly, strongly immunolabelled tangles were weakly thioflavine-stained. Secondly, less dense tangles were weakly immunolabelled but strongly thioflavine-stained. Thirdly, ghost tangles which correspond to extracellular NFT were exclusively thioflavine-stained. Thus, it is likely that NFT are degraded to form extracellular AF. Around neuritic plaques and some vessels with amyloid angiopathy, immunolabelled neurites, thioflavine-stained neurites and transition figures were also observed. On the other hand, the central core of plaques and pathological vessel walls were strongly thioflavine-stained but were never immunoreactive. In conclusion, these observations favour catabolism of PHF bundles found in NFT and in degenerating neurites into an amyloid substance. This amyloid substance seems different from other amyloid deposits found in the central core of neuritic plaques and vessel walls.

Diagnostic value of abdominal fat tissue aspirate in familial amyloid polyneuropathy

To confirm amyloid deposition we performed aspiration biopsy of abdominal fat tissue in 14 patients with type I familial amyloid polyneuropathy (FAP). All patients, about half of whom were at an early stage of disease and lacking severe neurological disabilities, showed positive amyloid deposition in preparations stained with Congo red. On light and electron microscopic examinations deposits of amyloid were observed around fat cells and small vascular walls, and among collagen bundles. All patients in the study were demonstrated to have a variant transthyretin in their sera by radio-immunoassay. Abdominal fat tissue aspiration can be easily performed with an ordinary syringe and is very sensitive for detecting amyloid deposition. The procedure is valuable in the diagnosis of type I FAP patients with actual deposition of amyloid substance, even at an early stage.