Isolation and characterization of a human heart cDNA encoding a new member of the small heat shock protein family--HSPL27.

A novel cDNA clone was isolated from a human adult heart cDNA library. This cDNA clone is similar to the small heat shock protein (smhsp) in both DNA and amino acid sequences, especially in the conserved region. Sequence analysis has shown that the putative novel smhsp, named 27 kDa heat-shock-protein-like protein (HSPL27) is a protein of 241 amino acids with a deduced molecular mass of 26.7 kDa and a deduced pI of 8.0. We have expressed the HSPL27 in E. coli and the expressed protein was found to be present in the soluble fraction of the bacterial cell lysate. Chromosomal mapping data shows that the HSPL27 gene is located at human chromosome 5q11.2.

A novel cDNA encoding a human homologue of ribosomal protein L29.

During the large scale partial sequencing of human heart cDNA clones, a novel clone which is very similar to the rat ribosomal protein L29 in both DNA and amino acid sequences was found. The cDNA encodes a protein with a deduced molecular weight of 17751 (159 aa). It shows 80.4% homology to protein L29 from the large ribosomal subunit of rat and is related to yeast YL43. The putative protein was named human ribosomal protein L29 (hRPL29). hRPL29 has a large excess of basic residues over acidic ones. The large amount of charged residues makes the protein very hydrophilic and the protein has a deduced pI of 12.16. Internal repeats have been characterised in many ribosomal proteins and a tandem repeat of KAKAKAKA was found to be unique to hRPL29. Analysis of gene organisation by Southern blotting shows that of the approximate 10 copies of hrpL29, all but one are pseudogenes. Northern analysis indicated that the mRNA that encodes human L29 is approx. 800 base pairs in length. An intron of hrpL29 has also been cloned and sequenced by polymerase chain reaction using human genomic DNA as the template.

Molecular cloning and characterization of FHL2, a novel LIM domain protein preferentially expressed in human heart.

A full-length cDNA clone encoding a novel LIM-only protein was isolated and sequenced from a human fetal heart cDNA library. This full-length clone consists of 1416 base pairs and has a predicted open reading frame (ORF) encoding 279 amino acids. The ORF of this polypeptide codes for the human heart-specific four and a half LIM-only protein 2 (FHL2). It possesses an extra zinc finger that is a half LIM domain and four repeats of LIM domain. When the human FHL2 cDNA probe was used to hybridize with poly-A RNA of various human tissues, a very strong signal could be seen in heart tissues, and only moderately low signals could be detected in placenta, skeletal muscle and ovary. Virtually no signal could be detected in brain, lung, liver, kidney, pancreas, spleen, thymus, prostate, testis, small intestine, colon or peripheral blood leukocyte. FHL2 was mapped to chromosome 2q12-q13 by fluorescent in-situ hybridization (FISH).

Articular cartilage transplantation.

This report describes the biopsy findings in four of 30 patients treated with cadaver osteochondral shell allografts for osteoarthritis in the knee. This study demonstrates that graft cartilage cells can survive in excess of 25 months, and that host bone can completely replace graft bone by creeping substitution. An inflammatory reaction in synovium and bone marrow was found in only one of four cases. Graft failure was related to prolonged down time of donor cartilage in one case and mechanical factors related to osteoarthritis in the apposing femoral surface in other cases. The clinical success of these grafts is attributed to the prolonged viability of cartilage cells, the capacity of host bone to join graft cartilage without histologic reaction, and the host's immunologic tolerance, which obviates the need for immunosuppressive therapy.

Effects of methylprednisolone and azathioprine on bronchial healing following lung autotransplantation.

The effect of low-dose immunosuppressive therapy upon the healing of the bronchial anastomosis and skin wound following lung autotransplantation was evaluated. Autotransplantation was performed in two groups of dogs: Group 1 (15 dogs) received no immunosuppression and Group 2 (13 dogs) received postoperative immunosuppression with methylprednisolone (2 mg/kg) and azathioprine (1.5 mg/kg). Two to four skin incisions 7 cm in length were made in the dorsal region of each dog. Dogs were put to death at 4, 9, 16, and 23 days postoperatively and the bronchial anastomoses and skin wounds were evaluated by breaking strength measurements. Bronchus and skin breaking strength increased with time in both groups. Bronchus breaking strength was similar in the two groups at day 4 and day 9. However, by day 23 bronchus breaking strength was significantly higher in Group 1 (p less than 0.001). Skin breaking strength was significantly higher in Group 1 on days 9 and 16 (p less than 0.005) and on day 23 (p less than 0.001). Our results suggest that low-dose methylprednisolone and azathioprine significantly affect the breaking strength of both bronchial anastomoses and healing skin incisions following canine lung autotransplantation.

Aggressive thyroid cancer in low-risk age population.

Seventy-eight patients under the age of 40 (low-risk patients) who had undergone surgical treatment for well-differentiated thyroid carcinoma were referred from 1979 to 1986 to our hospital for adjuvant therapy. A subgroup of 37 patients, 14 with apparent aggressive cancer, was studied. This study group consisted of 27 female and 10 male patients with mixed papillary and follicular cancer, who ranged in age from 11 to 40 years. Nodal disease occurred in 27 (73%) patients and invasiveness in 30 (81%) patients and involved multiple areas in 9 (24%) patients. Recurrence occurred in 14 (38%) patients and visceral metastases occurred in eight (22%) patients. All patients underwent appropriate surgery, but microscopic residual disease was seen in 15 patients and gross residual disease in seven patients, so that 31 patients underwent iodine-131 therapy, and 17 of these patients also underwent external radiation therapy. Three patients died of their disease, whereas 24 (65%) patients are free of disease and 9 (24%) patients are alive with disease. An additional 7 (19%) patients were initially seen in the fifth to seventh decade after decades of neglected thyroid disease, which culminated in residual cancer and death. Although low-risk categorization for thyroid cancer appears valid, its rigid application in support of conservative treatment may lead to inadequate primary treatment and underdiagnosis of cancer in thyroid nodule disease in the low-risk age population.

Induction of hyperparathyroidism by radioactive iodine.

External irradiation to the head and neck has been associated with the emergence of hyperparathyroidism, due both to adenoma and hyperplasia since a single case report was published in 1975. A recent report indicates that there may be some association between hyperparathyroidism and radioiodine administered for thyroid disease. Eight patients are presented herein, all female and in middle age, who presented with hyperparathyroidism due to adenoma (six patients) or hyperplasia (one patient who had undergone radioiodine treatment). In three patients, the dose of radioiodine exceeded 30 mCi. The duration of latency ranged from 4 to 20 years. Associated thyroid disease included cancer, colloid nodules, and thyroiditis. All patients became eucalcemic and have remained so. Experimental results that have supported the influence of radioiodine on parathyroid hyperactivity do not appear completely applicable to the human model. It is believed that the low mitotic rate equivalent to low radiosensitivity may have protected the parathyroid gland somewhat from neoplastic change, although the association of thyroid carcinoma and parathyroid adenoma appears to be a significant expression of radiation exposure. It may be the lack of a pituitary-stimulating hormone and the absence of an abscopal mechanism, as illustrated by irradiation to the thyroid, that explains the relative immunity of parathyroid behavior in the face of radioiodine exposure. Regardless of the frequency and validity of the association, it is important to recognize the possibility of the association illustrated by our case reports and scrutinize appropriate patients by intermittent serum calcium sampling, both for confirmation and possible detection of occult hyperparathyroidism.

Virus-induced osteosarcoma in rats.

An experimental model for osteosarcoma was developed in which, after Moloney murine sarcoma virus was injected into the tibial marrow space of three strains of inbred neonatal rats, a highly malignant neoplasm arose within ten days. This tumor was readily maintained in tissue culture and was transplantable to adult rats. It arose in the metaphyseal marrow of several bones, was locally invasive, metastasized to the lungs, and histologically resembled osteosarcoma.

Lipoma and liposarcoma of the parotid gland: high-resolution preoperative imaging diagnosis.

Over the past 7 years, nine fatty tumors within the parotid gland have been managed (eight lipomas, one liposarcoma). High-resolution computed tomography examination was carried out in all cases; with correct preoperative diagnosis recorded each time. The computed tomography imaging characteristics of lipoma, liposarcoma, and the differential diagnosis from other fat density lesions, such as a fatty infiltration, appear quite specific. The liposarcoma and six of the lipomas were resected at formal parotidectomy with facial nerve preservation. Two patients with small intraglandular lipomas have elected to undergo long-term clinical and imaging observation. Our experience indicates that high-resolution, soft-tissue imaging with computed tomography and magnetic resonance imaging permits consistent preoperative fatty tumor diagnosis. This imaging input facilitates rational treatment decision-making.

Campylobacter jejuni enterocolitis. A clinicopathologic study.

Sixteen patients with diarrhea due to Campylobacter jejuni seen within a one-year period at a general hospital were studied to review the clinical and pathological features of this illness. Campylobacter jejuni causes an acute diarrheal illness often associated with fever, delayed-onset hematochezia, and severe abdominal pain. Roentgenographically, one may see colonic and ileal ulceration. Sigmoidoscopically, the rectal appearance is similar to that from acute idiopathic ulcerative colitis, while rectal biopsy specimens show preservation of glandular architecture and a range of focal inflammatory changes. These changes are most severe in patients with a history of frank blood in stool, provided the specimens are taken within the first week of illness. No correlation between stool frequency, abdominal pain, or fever and the severity of proctitis in rectal biopsy specimens can be drawn, which suggests that the pathogenic determinants for thesse clinical manifestations may not be in the rectum, but higher in the colon or in the small intestine.

Brown bowel syndrome in Crohn's disease.

We describe a patient with bowel lipofuscinosis developing subsequent to Crohn's disease of five-years' duration. Lipofuscin pigment was demonstrated in the perinuclear region of smooth muscle cells with the longitudinal and circular coats of the ileal muscularis propria but not in the muscularis mucosa. Electron microscopy showed lipofuscin pigment forming within degenerating mitochondria, confirming the "brown bowel" syndrome as a smooth muscle mitochondrial myopathy.

The importance of extensive sampling and examination of cervix in suspected cases of amniotic fluid embolism.

Amniotic fluid embolism is an important complication of pregnancy with high mortality. The diagnosis of amniotic fluid embolism is generally made postmortem and rests upon the histological demonstration of amniotic fluid debris, including foetal epithelial squames and hair, in the pulmonary vasculature. We have made the diagnosis of amniotic fluid embolism in two patients by detection of the amniotic fluid debris in the blood vessels of the cervix in their hysterectomy specimens. These two patients presented with profuse primary postpartum haemorrhage and evidence of disseminated intravascular coagulation after uneventful deliveries. Amniotic fluid debris were only demonstrated in the blood vessels of the cervix but not in the corpus. This observation emphasizes the importance of a thorough histological examination of the cervix in cases of suspected amniotic fluid embolism.

Phagocytosis of colloidal carbon and heterologous red blood cells in the bone marrow of rats and rabbits.

Phagocytosis in the bone marrow of rats and rabbits was studied following intravenous injections of colloidal carbon and chicken red blood cells. In both animal species the marrow response to these different foreign particles was quite similar. There was an initial aggregation and degranulation of platelets around the injected particulate matter within the marrow sinuses. Then pseudopods of marrow macrophages appeared in the sinus lumen, forming a web-like structure which trapped and phagocytosed the injected foreign material as well as the platelets. Within the phagosomes of these macrophages, the injected material and platelets were withdrawn into the parenchyma, where degradation or storage occurred. This sequence of events suggests that platelets may play an important role in marrow phagocytosis. The most active cells in marrow phagocytosis are the macrophages. The endothelial cells participated in the phagocytosis of colloidal carbon. The amount of carbon within these cells, however, was small in comparison with that trapped by macrophages. Further, the endothelial cells did not phagocytose chicken red blood cells. These results, being similar to those obtained in lymph, spleen and liver, challenge the concept of the reticuloendothelial system. The term of macrophage system is proposed as a replacement.

Ultrastructure of the normal lymph node.

The "normal" lymph node has been studied by electron microscopy. The lymphoid tissue can be divided into three distinct zones. Zone 1 consists of loosely arranged cells surrounding the lymphatic sinuses and blood vessels. This is the only zone in which plasma cells are present. Zone 2 is surrounded by zone 1 and consists of compactly arranged cells in which lymphocytes predominate. Zone 3 (germinal center) appears only after antigenic stimulation. It is characterized by large, ribosome-rich cells and macrophages containing phagocytosed lymphocytes. These zones are arranged with their longest diameters pointing towards the hilus. Zone 1 is the longest and extends across the cortex, paracortex and medulla. Zone 2 spans across cortex and paracortex. Zone 3 usually is confined to the cortex. Our preliminary studies indicate that zone 1 is mainly bursal dependent, zone 2 is mainly thymic dependent and zone 3 is bursal dependent.

Complement-mediated pulmonary edema in sheep.

Incubation of plasma with zymosan results in complement activation. Infusion of zymosan-activated plasma (ZAP) into the pulmonary circulation in sheep results in a transient rise in pulmonary vascular resistance, accompanied by hypoxemia. This is associated with production of the vasoactive prostaglandin metabolite, thromboxane. We hypothesized that ZAP infusion caused pulmonary edema and may transiently alter pulmonary permeability to protein. Two sets of experiments were conducted involving six sheep. Three sheep underwent open lung biopsies during ZAP infusion, and electron microscopy documented early reversible interstitial pulmonary edema in response to ZAP infusion. Three other sheep were prepared with lung lymph fistulae and subjected to a similar infusion of ZAP. These studies documented a rapid rise in lymph flow and lymph protein clearance, occurring immediately upon infusion of ZAP. These experiments demonstrate a rapid reversible interstitial edema in sheep lung in response to ZAP infusion and suggest that permeability to protein may be transiently altered as a result of this injury.

Xenogeneic red blood cell degradation in a regional lymph node and dissemination of antigens by circulating lymphocytes.

The degradation of intramuscularly injected autogenic and xenogenic red blood cells are studied with electron microscopy and immunoferritin technique. Autogenic red blood cells are phagocytosed and completely degradated by macrophages within the draining lymph node in a relatively short time. Phagocytosis and initial degradation of xenogeneic red blood cells by macrophages in the draining lymph node are similar; however, portions of the degradation products persist in macrophages as characteristic myelinic-like figures. After 4 days, identical figures appear in large numbers in the adjacent lymphocytes within membrane-bound vacuoles. With the conjugated ferritin antibody technique, these myelinic-like figures are shown to contain antigens of the xenogeneic red blood cells. Subsequently, lymphocytes containing the inclusions are found in the peripheral blood, the marginal zone of the spleen, other lymph nodes, and the bone marrow. As in the draining lymph node the number of plasma cells increases in these organs in areas adjacent to lymphocytes with the inclusions. In a parallel study, uncoated polystyrene spheres injected intramuscularly accumulate and remain localized in macrophages of the draining lymph node. When coated with xenogeneic hemoglobin and membranes, these spheres appear in lymphocytes of the draining lymph node and subsequently in lymphocytes of the blood and spleen.