A novel SERPINA12 variant and first European patients with diffuse palmoplantar keratoderma.

BACKGROUND: Hereditary palmoplantar keratodermas (hPPKs) comprise a heterogeneous group of skin disorders characterized by persistent palmoplantar hyperkeratosis. Loss-of-function variants in a serine peptidase inhibitor, SERPINA12, have recently been implicated in autosomal recessive diffuse hPPK. The disorder appears to share similarities with another hPPK associated with protease overactivity, namely Nagashima-type PPK (NPPK) caused by biallelic variants in SERPINB7. OBJECTIVES: The aim of this study was to enhance the understanding of the clinical and genetic characteristics of serine protease-related hPPKs caused by variants in SERPINA12 and SERPINB7. METHODS: Whole-exome sequencing (WES) was performed for hPPK patients. Haplotype analysis was completed for the patients with identified recessive SERPINA12 variants and their available family members. In addition, the current literature of SERPINA12- and SERPINB7-related hPPKs was summarized. RESULTS: The phenotype of SERPINA12-related hPPK was confirmed by reporting three new SERPINA12 patients, the first of European origin. A novel SERPINA12 c.1100G>A p.(Gly367Glu) missense variant was identified confirming that the variant spectrum of SERPINA12 include both truncating and missense variants. The previously reported SERPINA12 c.631C>T p.(Arg211*) was indicated enriched in the Finnish population due to a plausible founder effect. In addition, SERPINA12 hPPK patients were shown to share a similar phenotype to patients with recessive variants in SERPINB7. The shared phenotype included diffuse transgradient PPK since birth or early childhood and frequent palmoplantar hyperhidrosis, aquagenic whitening and additional hyperkeratotic lesions in non-palmoplantar areas. SERPINA12 and SERPINB7 hPPK patients cannot be distinguished without genetic analysis. CONCLUSIONS: Recessive variants in SERPINA12 and SERPINB7 leading to protease overactivity and hPPK produce a similar phenotype, indistinguishable without genetic analysis. SERPINA12 variants should be assessed also in non-Asian patients with diffuse transgradient PPK. Understanding the role of serine protease inhibitors will provide insights into the complex proteolytic network in epidermal homeostasis.

A novel desmoplakin mutation causes dilated cardiomyopathy with palmoplantar keratoderma as an early clinical sign.

BACKGROUND: PPKs represent a heterogeneous group of disorders with hyperkeratosis of palmar and/or plantar skin. PPK, hair shaft abnormalities, cardiomyopathy and arrhythmias can be caused by mutations in desmosomal genes, e.g. desmoplakin (DSP). PPK should trigger genetic testing to reveal mutations with possible related cardiac disease. OBJECTIVES: To report a large multigenerational family with a novel DSP mutation associated with early-onset PPK and adult-onset cardiomyopathy and arrhythmias. METHODS: A custom-designed in-house panel of 35 PPK related genes was used to screen mutations in the index patient with focal PPK. The identified DSP mutation was verified by Sanger sequencing. DNA samples from 20 members of the large multigenerational family were sequenced for the DSP mutation. Medical records were reviewed. Clinical dermatological evaluation was performed, including light microscopy of hair samples. Cardiac evaluation included clinical examination, echocardiography, cardiac magnetic resonance imaging (CMR), electrocardiogram (ECG), Holter monitoring and laboratory tests. RESULTS: We identified a novel autosomal dominant truncating DSP c.2493delA p.(Glu831Aspfs*33) mutation associated with dilated cardiomyopathy (DCM) with arrhythmia susceptibility and focal PPK as an early cutaneous sign. The mutation was found in nine affected family members, but not in any unaffected members. Onset of dermatological findings preceded cardiac symptoms which were variable and occurred at adult age. CONCLUSIONS: We report a novel truncating DSP mutation causing focal PPK with varying severity and left ventricular dilatation and ventricular extrasystoles. This finding emphasizes the importance of genetic diagnosis in patients with PPK for clinical counselling and management of cardiomyopathies and arrhythmias.

Hereditary palmoplantar keratoderma - phenotypes and mutations in 64 patients.

BACKGROUND: Hereditary palmoplantar keratodermas (PPK) represent a heterogeneous group of rare skin disorders with epidermal hyperkeratosis of the palms and soles, with occasional additional manifestations in other tissues. Mutations in at least 69 genes have been implicated in PPK, but further novel candidate genes and mutations are still to be found. OBJECTIVES: To identify mutations underlying PPK in a cohort of 64 patients. METHODS: DNA of 48 patients was analysed on a custom-designed in-house panel for 35 PPK genes, and 16 patients were investigated by a diagnostic genetic laboratory either by whole-exome sequencing, gene panels or targeted single-gene sequencing. RESULTS: Of the 64 PPK patients, 32 had diffuse (50%), 19 focal (30%) and 13 punctate (20%) PPK. None had striate PPK. Pathogenic mutations in altogether five genes were identified in 31 of 64 (48%) patients, the majority (22/31) with diffuse PPK. Of them, 11 had a mutation in AQP5, five in SERPINB7, four in KRT9 and two in SLURP1. AAGAB mutations were found in nine punctate PPK patients. New mutations were identified in KRT9 and AAGAB. No pathogenic mutations were detected in focal PPK. Variants of uncertain significance (VUS) in PPK-associated and other genes were observed in 21 patients that might explain their PPK. No suggestive pathogenic variants were found for 12 patients. CONCLUSIONS: Diffuse PPK was the most common (50%) and striate PPK was not observed. We identified pathogenic mutations in 48% of our PPK patients, mainly in five genes: AQP5, AAGAB, KRT9, SERPINB7 and SLURP1.

Prediction of survival in breast cancer: evaluation of different multivariate models.

BACKGROUND: We compared the performance of multivariate models based on mitotic activity index, lymph node status, and tumor size in the prognostication of breast cancer. MATERIAL AND METHODS: Cox and discriminant models for survival were created for two patient groups: a) 120 breast cancer patients, and b) 86 patients with ductal infiltrating carcinoma. The models were compared with the model of Baak et al (1985). RESULTS: The models distinguished between dying and surviving patients with an efficiency of 70.9-77.9% in mutual tests. With a single cutoff the model of Baak et al was less efficient (50.8-65.8%). If a region of uncertainty was allowed between two cutpoints, the efficiencies below and above the cutpoints increased. When the uncertain region included a third of the patients, the efficiency varied between 73.8 and 84.7%. CONCLUSION: Multivariate models seem to need a region of uncertainty between two discriminating cutpoints. These models resulted in the correct prediction of prognosis in about 75% and more of patients. With different materials the models differed in efficiency. With a region of uncertainty the model of Baak et al performed well with completely independent material.

Maternal drug treatment and human milk banking.

We studied prospectively during one year temporary drug use by mothers donating breast milk to assess the problem of drug treatment of donors. Sixty-four of the 284 mothers (22.5%) had to abstain from donating due to medication. The indication was infection in 50/56 treatments (89.3%). Antimicrobial agents were prescribed 44/52 times (84.6%). The channelling of milk from mothers in early phases of lactation to premature and newborn infants was identified as a special risk situation, if mothers on medication are not excluded. The limited number of such donors leads to use of milk unpooled or pooled to small volumes with increased risk for adverse effects to babies as a consequence. We recommend a wash-out period of 5 half-lives of the drug after the last ingested dose. For the majority of drugs in this study, with some important exceptions, a wash-out period of 1 day was sufficient.

Prognostic studies in breast cancer. Multivariate combination of nodal status, proliferation index, tumor size, and DNA ploidy.

We studied histological samples and clinical data from 111 breast carcinoma patients originally treated in 1975-1977 who did not have distant metastases at the time of diagnosis. A multivariate survival analysis using the Cox model selected the studied variables in the following order according to their prognostic association (breast cancer deaths, or deaths from any cause): axillary lymph node status, tumor size, mitotic index, and DNA ploidy status. The association of the different variables to the prognosis in respect to breast cancer deaths was evaluated 1-10 years after treatment by stepwise logistic regression. Lymph node status, tumor size, mitotic index, and DNA ploidy all showed significant relation to the prognosis but this association varied considerably with time of observation.

Prevalence of familial hypercholesterolemia among young north Karelian patients with coronary heart disease: a study based on diagnosis by polymerase chain reaction.

Two deletions of the low density lipoprotein (LDL) receptor gene account for about 90% of the mutations that cause familial hypercholesterolemia (FH) in eastern Finland. The FH-Helsinki mutation deletes exons 16, 17 and a portion of exon 18, while the FH-North Karelia allele is characterized by a deletion of seven nucleotides from exon 6 of the LDL receptor gene. We developed a DNA assay based on the use of polymerase chain reaction (PCR) which simultaneously detects both of these mutations. We have screened 90 young (< 45 years) eastern Finns with symptomatic coronary heart disease (CHD) for the presence of these FH genes. One or the other of the mutations was present in 4 out of 55 survivors of acute myocardial infarction (AMI) and 4 out of 35 patients with angina pectoris (AP), but in none of 50 healthy controls of similar age. These data show a relatively high prevalence of confirmed FH in young CHD patients (AMI and MI combined: 8/90, or 9%), and also demonstrate the feasibility of PCR techniques in diagnosis of FH among populations with enrichment of specific types of LDL receptor gene mutations.

Thromboprophylaxis in patients with hip fractures: a prospective, randomized, comparative study between Org 10172 and dextran 70.

A prospective, randomized, assessor-blind trial has been undertaken to compare the thromboprophylactic effect and safety of the heparinoid Org 10172 (a mixture of low molecular-weight sulfated glycosaminoglycuronides) and dextran 70 in patients operated on for hip fracture. Prestudy biostatistical calculations led to the need for 260 patients. Three hundred eight patients were randomized and 19 were excluded after randomization, the majority because of postponed surgery. Analyses were made on the 289 patients on an intention-to-treat basis, as well as on the 247 patients given correct prophylaxis. Diagnosis of deep vein thrombosis was based on bilateral ascending phlebography on postoperative days 10 through 12. The frequency of deep vein thrombosis on an intention-to-treat basis was 10% in the Org 10172 group and 30% in the dextran 70 group and, on the basis of correct prophylaxis, 12% and 31%, respectively, both differences being significant (p less than 0.001). Two-month mortality rates were equal in the groups. Three fatal pulmonary emboli were seen in the dextran group. Significantly more patients in the dextran group received postoperative transfusions; no other differences in various hemorrhagic parameters were seen. Thus it can be concluded that Org 10172 has a significantly better thromboprophylactic effect than does dextran in patients with hip fractures without significant side effects.

The use of fibrinogen uptake test in screening for deep vein thrombosis in patients with hip fracture.

255 hip fracture patients were studied by 125I-fibrinogen uptake test and bilateral phlebography. We found the sensitivity of fibrinogen scanning to be 44% for the non-operated limb and 50% for the calves. The predictive value of a negative result was found to be 92% and 93% respectively. We conclude that the use of fibrinogen uptake test as single diagnosticum is not valid and can only be recommended in combination with phlebography when studying patient where the frequency of DVT is expected to be low.

An improved prognostic index of axillary node involvement in breast cancer incorporating DNA ploidy and tumour size.

A multivariate prognostic index based on clinical data and the results of flow cytometry for the grading of breast cancer was evaluated in 117 patients whose disease had been detected and treated by mastectomy with axillary clearance between 1974 and 1976. Survival analysis with Cox's regression model pointed to three important prognostic factors: lymph node involvement (p less than 0.001), DNA ploidy (p less than 0.01) and tumour size (p less than 0.01). These factors were incorporated into a prognostic index, in which the lymph node involvement, DNA ploidy, and tumour size contributed to the index in that order. Logistic discriminant analysis with five year follow-up as the fixed end point (70 alive, 47 dead) gave the same result; lymph node involvement, tumour size, and DNA ploidy were the best prognostic indicators of survival. The result showed that our multivariate prognostic index was more powerful than lymph node involvement alone. The use of this prognostic index is recommended for selecting patients for different treatments.

Relationship between DNA ploidy and survival in patients with primary breast cancer.

The DNA ploidy of breast cancer tissue from paraffin blocks was measured by flow cytometry in 117 patients whose disease had been detected and treated with surgery between 1974 and 1976. Patients with aneuploid tumours had positive axillary nodes and distant metastases more often than those with diploid tumours. Aneuploid tumours were more common in postmenopausal than premenopausal women. The S-phase fraction (SPF) was significantly higher in aneuploid than in diploid tumours and positive axillary lymph nodes were found in 26 per cent of the patients who had a tumour with a SPF below the median (4.8 per cent) and in 48 per cent of those with tumours with SPF values above the median. At the primary clinical investigation 2 per cent of the patients with diploid tumours and 6 per cent of those with aneuploid tumours had distant metastases. During the follow-up, the proportion of patients with distant metastases increased to 42 and 72 per cent, respectively. With a follow-up of 11.5 years, the DNA aneuploidy of the tumour showed a significant association with decreased survival. Thirty-three per cent of patients with diploid and 65 per cent of patients with aneuploid tumours had died from breast cancer during the follow-up (P less than 0.001). All patients with hypertetraploid or multiploid tumours died from breast cancer. High SPF values were associated more closely with distant metastases or death during the follow-up than low SPF values. Our results suggest that DNA ploidy measured by flow cytometry from paraffin embedded tissue blocks of human breast cancer can be used to predict the aggressiveness of the tumour and the survival of the patients.

Iatrogenic compartment syndrome, A follow-up of four cases caused by elastic bandage.

We describe four patients suffering from lower limb compartment syndromes which were caused by constrictive bandages applied after stripping of varices. The dressing was erroneously only partially removed, when the patients started complaining of severe pain and tension in the operated legs. The damages varied from extended irreversible neuromuscular defects to lesser functional handicaps. Three patients had corrective surgery. The clinical follow up over several years showed little improvement, secondary complaints were frequent.

Diet, serum, breast fluid growth hormone, and prolactin levels in normal premenopausal Finnish and American women.

A food frequency questionnaire was used to estimate and compare the dietary fat and fiber consumption of 94 premenopausal women in Kuopio (rural Finland, where there is a relatively low risk of breast cancer) and 61 American women in New York (where there is a high risk of breast cancer). In keeping with previous reports concerning middle-aged men, both groups had high fat intakes, but the Finnish women had considerably higher fiber intakes (24 +/- 11 vs. 16 +/- 6 g). Serum and breast fluid growth hormone and prolactin levels were compared in 29 of these American women and 24 of the Finnish women. All were healthy and had regular menstrual cycles. Serum growth hormone levels, which were measured by radioimmunoassay, were higher in the Finnish women; all but three of their breast fluids contained detectable growth hormone, frequently at extremely high concentration. In contrast, only 2 of the 29 breast fluids from American women had detectable amounts of growth hormone. Of the Finnish samples, 10 were also measured by an immunoradiometric assay with high specificity for the 22,000-dalton growth hormone molecule; all but 3 had values less than 3.0 ng/ml. Serum and breast fluid prolactin concentrations, which were determined by radioimmunoassay, were no different in the two groups; both groups frequently had considerably higher levels in breast fluid compared with the corresponding serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum and breast duct fluid prolactin and estrogen levels in healthy Finnish and American women and patients with fibrocystic disease.

Nipple aspiration yielded measurable amounts of breast duct fluid for 27 of 42 (64%) healthy premenopausal Finnish women, 93 of 218 (43%) premenopausal American patients with fibrocystic disease, but only 24 of 92 (26%) healthy American premenopausal controls. When aspiration was successful, the average volumes obtained were larger for the normal Finnish women and, particularly, for the American fibrocystic disease patients compared with the American controls. The difference in secretion between the healthy Finnish and American women appeared to be related to a history of breast feeding, and its duration. While serum estrogen and prolactin concentrations were similar in the three groups, prolactin levels in breast fluids from the Finnish women were frequently higher than those in the American controls. A similar trend, which did not reach statistical significance, was observed in the 43 of 93 (46%) secretors with fibrocystic disease and cyclical mastalgia.

The 125I-labelled fibrinogen uptake test in elective hip surgery.

The value of the 125I-labelled fibrinogen uptake test as a screening method for deep vein thrombosis was estimated in 49 patients (97 extremities) undergoing elective hip operations. The legs were monitored preoperatively, on the 1st, 2nd and 4th postoperative days. Contrast venography, as a reference method, was performed on the 5th postoperative day. The sensitivity was 73%, specificity 85%, positive predictive value 63%, negative predictive value 86% and correlation coefficient 55%. 78% of fibrinogen uptakes appeared within the first 24 hours, and 6% on the 4th postoperative day. Only 1 of 3 patients with a positive fibrinogen uptake test in the femoral region had roentgenologic evidence of deep vein thrombosis. According to these results the isotope-labelled fibrinogen uptake test can also be utilized as a screening method in elective hip surgery.

A randomized study of a semisynthetic heparin analogue and heparin in prophylaxis of deep vein thrombosis.

The prophylactic effect of a semisynthetic heparin analogue (SSHA) on deep vein thrombosis was investigated in a prospective double-blind multicentre trial. 440 major general surgical and gynaecological patients were randomized to one of three treatment groups: 50 mg SSHA, 37.5 mg SSHA and 5000 units sodium heparin subcutaneously 12-hourly. Deep venous thrombosis (DVT) was diagnosed with the fibrinogen uptake test and verified with phlebography. Bleeding complications and other side-effects were carefully monitored. There were no significant differences between the three treatment groups of patients in age, sex, type of operation or risk factors. A DVT was diagnosed in 16 patients (12 per cent) in the SSHA 50 mg group, in 21 patients (15 per cent) in the SSHA 37.5 mg group and 21 patients (14 per cent) in the heparin-treated group. No significant differences were found in the number of patients who bled unexpectedly in the postoperative period, required transfusion or developed wound haematomas. Blood loss at operation was similar in all three groups. Three pulmonary emboli were diagnosed by pulmonary scintigraphy, one in each group.

Screening for postoperative pulmonary embolism on the basis of clinical symptomatology, routine electrocardiography and plain chest radiography.

The incidence of pulmonary embolism (PE) and the usefulness of various screening methods for its detection were studied in 108 patients undergoing elective hip surgery. Twenty patients had PE as shown by perfusion-ventilation lung scintigraphy. Six of them had symptoms of embolism. Clinical symptoms of PE, highly suggestive electrocardiographic (ECG) signs and signs in chest radiographs showed high specificity but low sensitivity with regard to PE. If the non-specific ECG signs and the symptoms and signs of deep vein thrombosis in the calves had also been taken into account, it would have been possible to identify 95% of the patients with PE. The use of all these screening tests revealed a suspicion of PE in two thirds of the patients. The simplest way to screen for postoperative PE seems to be to use a combination of the symptoms and signs of both PE and deep vein thrombosis, and to identify any tachycardia. This method gave a sensitivity of 85%.

Computed tomography of spinal and pelvic fractures.

The value of computed tomography (CT) compared with conventional x-ray was assessed in 29 spinal and 9 pelvic fractures. In the so called bursting fractures of spinal vertebrae the position of bone fragments in the neural canal was shown better by CT. The neural canal was remarkably narrowed on several occasions without the patient having any clinical symptoms of root or medullary compression. In 2 patients a fracture of laminae was discernible only on CT. Vertical fractures of vertebral bodies were also difficult to distinguish on the conventional plain x-ray. CT was helpful for planning of the anterolateral decompression operation of L2 in one patient. In cases of pelvic trauma sacro iliac joint diastasis was readily shown by CT. CT was clearly better able to determine whether the fracture extended across the sacro iliac joint. Also in fractures of the iliac wings CT readily showed the internal and external surfaces which were difficult to evaluate on plain radiographic examination. In spinal and pelvic fractures CT can give useful additional information which can influence the treatment of the trauma patient.