Tibial Nerve Block as Treatment of Chronic Foot Pain.

Background: Minimally invasive therapies can alleviate pain and improve walking in individuals with persistent foot and ankle pain. Objectives: The current study aimed to define the protracted consequences of tibial nerve blocks with steroids for individuals with persistent foot pain and to investigate the link between the thermography of the plantar foot and the beneficial effect of a tibial nerve block with steroids. Methods: All patients with chronic foot pain (n = 45) in this cohort underwent a block of the tibial nerve in the Department of Pain Therapy of Pain Clinic De Bilt, Utrecht, Netherlands, within November 2019 to April 2020. The thermographic images of patients were taken before and after injection. Results were retrospectively evaluated after 18 months. Results: In this study, 53% of the patients had pain relief at 7 weeks of follow-up with a unilateral or bilateral block of the tibial nerve. An improvement in walking distance was reported by 22% of the patients. Side effects of the tibial nerve block reported at 7 weeks of follow-up increased pain (5%) and the occurrence of leg cramps (5%) among the treated patients. At 18 months, 45% of the successfully treated feet still had benefits. A difference between the big toe's temperature and the foot's average temperature of less than -0.9°C on thermography before and after the tibial nerve block can predict a beneficial result of therapy. Conclusions: Tibial nerve block provides a safe, minimally invasive treatment option for almost half of the patients with painful feet in this cohort, and when successful, it can last a long term. Thermographic imaging of the plantar foot can predict only to a small extent the beneficial effect of the tibial nerve block with steroids on foot pain. Tibial nerve block should be considered when custom foot orthoses have been inadequate for pain relief or restricted walking distance.

Genetic Lesions in Russian CLL Patients with the Most Common Stereotyped Antigen Receptors.

Chronic lymphocytic leukemia (CLL) is one of the most common B-cell malignancies in Western countries. IGHV mutational status is the most important prognostic factor for this disease. CLL is characterized by an extreme narrowing of the IGHV genes repertoire and the existence of subgroups of quasi-identical stereotyped antigenic receptors (SAR). Some of these subgroups have already been identified as independent prognostic factors for CLL. Here, we report the frequencies of TP53, NOTCH1, and SF3B1 gene mutations and chromosomal aberrations assessed by NGS and FISH in 152 CLL patients with the most common SAR in Russia. We noted these lesions to be much more common in patients with certain SAR than average in CLL. The profile of these aberrations differs between the subgroups of SAR, despite the similarity of their structure. For most of these subgroups mutations prevailed in a single gene, except for CLL#5 with all three genes affected by mutations. It should be noted that our data concerning the mutation frequency in some SAR groups differ from that obtained previously, which could be due to the population differences between patient cohorts. The research in this area should be important for better understanding the pathogenesis of CLL and therapy optimization.

Reduction of the Quality of Contraceptive Counseling (QCC) scale to a short version (QCC-10) in Ethiopia, India, and Mexico.

OBJECTIVE: To reduce the Quality of Contraceptive Counseling (QCC) scale to a shortened version, coined the QCC-10, for use in measuring client-reported quality of counseling across varied settings. STUDY DESIGN: Secondary psychometric analysis of data collected for validating full versions of the QCC scale (QCC-Mexico, QCC-Ethiopia, QCC-India) and expert voting to reduce the original 26 QCC items to a 10-item set. RESULTS: Exploratory factor analysis revealed a clear one-factor solution for the QCC-10 in each country. Factor loadings were consistently >0.4 for all but two items; both were retained due to their importance to content validity. Internal consistency reliability of the QCC-10 was α=0.8 in Mexico and Ethiopia, and α=0.5 in India. QCC-10 scores were highly and positively correlated with a dichotomous overall measure of client experience and intention to initiate selected method, indicating convergent validity. CONCLUSION: The QCC-10 offers an innovative, cross-cultural approach to measuring quality in contraceptive counseling. Future efforts should examine its validity and reliability for use globally, with additional exploration of how to best measure negative aspects of care, particularly in India where such items were problematic. Thoughtful, nuanced measurement of client perspectives on their counseling experiences, available via the QCC-10, is critical to monitoring and improving quality of person-centered care and the fulfilment of human rights in contraceptive services worldwide. IMPLICATIONS: Cross-cultural, person-centered measures of quality in contraceptive counseling, such as the QCC-10, can help inform efforts to improve quality of family planning services and fulfillment of human rights. Future work will continue to explore the validity of this 10-item measure for use in various settings.

Repertoire of Rearranged Immunoglobulin Heavy Chain Genes in Russian Patients With B-Cell Lymphoproliferative Diseases.

INTRODUCTION: Immunoglobulin heavy chain variable region (IGHV) repertoire narrowing could be an evidence for the involvement of a limited set of antigens in the development of lymphomas. For chronic lymphocytic leukemia (CLL) the existence of more than 200 subgroups of tumor IGHV antigen-binding sites, so called "stereotypical" antigen receptors (SAR) has been shown. For others lymphomas the possibility of SARs is also suggested. The aim of this study is to compare the tumor IGHVs and possible SARs in various B-cell malignancies in Russia and other countries. MATERIALS AND METHODS: The study included samples of 1800 CLL patients, 52 patients with mantle cell lymphoma, 48 patients with hairy cell lymphoma and 37 patients with splenic marginal cell lymphoma. The nucleotide sequences of the IGHV genes were determined according to ERIC protocol. RESULTS: In CLL most common IGHV genes were IGHV1-69, IGHV1-2, IGHV3-30 and IGHV4-34. The most common SARs were CLL#1, CLL#6, CLL#2, CLL#3. In MCL the most common genes were IGHV4-34, IGHV3-21, IGHV3-23. In 5 MCL patients CDR3 sequences were identified matching definitions of a stereotyped. In the half of SMZL patients was identified gene IGHV1-2. Other IGHV genes were much less common. Two pairs of SMZL patients have motives similar to each other. In HCL IGHV repertoire was the most variable, no trends for antigen receptor stereotypy were observed. It was found that SARs are highly disease-specific both at the level of nucleotide and amino acid sequences. CONCLUSION: Our results suggest that antigens crucial for the pathogenesis of B-cell malignancies could be disease-specific. Further studies on extended samples of non-CLL patients concerning the role of SARs in pathogenesis of these diseases may also contribute to the development of new diagnostic and prognostic markers.

Sometimes they come back: New and old spinal muscular atrophy adults in the era of nusinersen.

BACKGROUND AND PURPOSE: Following the commercial availability of nusinersen, there have been a number of new referrals of adults with spinal muscular atrophy (SMA) not regularly followed in tertiary-care centers or enrolled in any disease registry. METHODS: We compared demographics and disease characteristics, including assessment of motor and respiratory function, in regularly followed patients and newcomers subdivided according to the SMA type. RESULTS: The cohort included 166 adult patients (mean age: 37.09 years): one type I, 65 type II, 99 type III, and one type IV. Of these 166, there were 67 newcomers. There was no significant difference between newcomers and regularly followed patients in relation to age and disease duration. The Hammersmith Functional Motor Scale Expanded and Revised Upper Limb Module scores were higher in the regularly followed patients compared to newcomers in the whole cohort and in both SMA II and II. A difference was also found on ventilatory status (p = 0.013) and Cobb's angle >50° (p = 0.039) between the two subgroups. No difference was found in scoliosis surgery prevalence (p > 0.05). CONCLUSIONS: Our results showed differences between the two subgroups, even if less marked in the type III patients. In the type II patients, there was a higher proportion of newcomers who were in the severe end of the spectrum. Of the newcomers, only approximately a third initiated treatment, as opposed to the 51% in the regularly followed patients. The identification of patients who were not part of the registries will help to redefine the overall prevalence of SMA and the occurrence of different phenotypes.

Cardiovagal regulation and transcutaneous pO2 in breast cancer patients - a pilot study.

Vagal activity in patients with metastatic or recurrent breast cancer can predict their survival and it can be altered by behavioral, pharmacological and surgical interventions. Tumor oxygenation is important in defining the cellular metabolic microenvironment of human malignancies, O2-depleted areas coincide with nutrient and energy deprivation and with a hostile metabolic microenvironment. In our work, we simultaneously measured two oxygen-sensitive parameters in breast cancer patients; blood oxygen saturation (SpO2) and trans-cutaneous O2 partial pressure (tcpO2) in breast tissue. Concurrently, 5-minute beat-to-beat heart rate recording was carried out in order to get heart rate variability (HRV) data from time-domain analyses, frequency-domain analyses and entropy and symbolic dynamic non-linear methods. We compared these parameters in patients newly diagnosed with breast cancer, in patients after therapy and in healthy controls. We found lower tcpO2 in patients with presence of malignant tumor compared to those post-treatment and/or without presence of malignancy. We also detected lower 2UV% (two unlike variations) and entropy in non-linear HRV analysis in all breast cancer patients and these parameters associated with parasympathetic activity did not return to the values comparable with healthy individuals after anti-cancer therapy, contrary to tcpO2. Our findings show that breast tissue tcpO2 can recover after the anti-cancer treatment, but complex heart rate control and cardio-vagal regulation remain impaired. This supports the idea that cancer patients and survivors might benefit from non-pharmacological interventions aimed at enhancing vagal activity, such as HRV biofeedback or Yoga.

Marital status is a prognostic factor in amyotrophic lateral sclerosis.

BACKGROUND AND OBJECTIVES: Several variables have been linked to a shorter survival in patients with amyotrophic lateral sclerosis (ALS), for example, female sex, older age, site of disease onset, rapid disease progression, and a relatively short diagnostic delay. With regard to marital status, previous studies suggested that living with a partner might be associated to a longer survival and a higher likelihood to proceed to tracheostomy. Therefore, to further strengthen this hypothesis, we investigated the role of marital status as a prognostic variable in a cohort of ALS patients. METHODS: We performed a retrospective analysis on 501 consecutive ALS patients for which a complete disease's natural history and clinical/demographic data were available. At diagnosis, 409 patients (81.6%) were married or lived with a stable partner, whereas 92 patients (18.4%) were single/widowed/divorced. RESULTS: In our ALS cohort, being married was associated with a median longer survival (married, 35 months [24-50] vs unmarried, 27 months [18-42]; P<.004). Moreover, married and unmarried patients were significantly different in many clinical and demographic variables, including age at disease onset, gender, body mass index, and number of children. Cox regression analysis showed that age at onset, diagnostic delay, and marital status were independent predictors of survival. In unmarried patients, female sex was also significantly associated with shorter survival. CONCLUSIONS: Marital status is a prognostic factor in ALS, and it significantly affects survival.

REM sleep behavior disorder and periodic leg movements during sleep in ALS.

OBJECTIVE: To assess sleep characteristics and the occurrence of abnormal muscle activity during sleep, such as REM sleep without atonia (RSWA), REM sleep behavior disorder (RBD), and periodic leg movements during sleep (PLMS), in patients with amyotrophic lateral sclerosis (ALS). METHODS: A total of 41 patients with ALS and 26 healthy subjects were submitted to clinical interview and overnight video-polysomnography. RESULTS: A total of 22 patients with ALS (53.6%) reported poor sleep quality. Polysomnographic studies showed that patients with ALS had reduced total sleep time, increased wakefulness after sleep onset, shortened REM and slow-wave sleep, and decreased sleep efficiency, compared to controls. Polysomnographic abnormalities were not different in patients reporting good or poor sleep and were not correlated to clinical and demographic variables. The PLMS index was significantly higher in patients with ALS than in healthy subjects, and 22 patients (53.6%) showed a PLMS index > 15/h, vs 4 (15.4%) controls (P < 0.001). Finally, two patients with ALS (4.9%) had RBD, and two more patients presented RSWA (4.9%), whereas no controls showed abnormalities of REM sleep. CONCLUSION: Patients with ALS frequently present abnormalities of sleep that can be documented both at the clinical interview and at the polysomnographic evaluation, including insomnia, fragmented sleep, and increased PLMS. Moreover, abnormalities of REM sleep can be found in some of these patients.

Evaluation of expression profiles of microRNAs and two target genes, FOXO3a and RUNX2, effectively supports diagnostics and therapy predictions in breast cancer.

Breast cancer (BC) including its progression into bone metastasis is a complex process involving changes in gene expression and function of both, microRNAs (miRNAs) and their target genes. Deregulation of miRNAs has been described as a crucial factor responsible for the initiation and progression of BC, and specific miRNA expression profiles have been found to be associated with particular disease states, histological tumor types, and BRCA1/2 or HER status. BRCA1 tumor suppressor is involved in DNA damage response and repair and epigenetically controls miR-155 expression and its pre-cancerous potential. MiR-155 targets 3´UTR region of multiple components of the pro-oncogenic signaling cascades, including FOXO3a tumor suppressor and RUNX2 transcription factor regulating metastatic potential in BC. We employed qRT-PCR to determine expression level and examine possible regulatory role of selected miRNAs (miR-17, miR-18a, miR-19a, miR-20a, miR-21, miR-27a and miR-155) and their impact on expression modulation of FOXO3a and RUNX2 in peripheral blood mononuclear cells (PBMCs) in healthy individuals, in women carrying BRCA1 mutations with no disease manifestation, in women carrying BRCA1 mutations after tumor resection and therapy and in women with BC of unknown BRCA1 status in acute stage before tumor resection. Our results showed significant increase of miR-17, miR-19a, miR-21, miR-27, miR-155 and RUNX2 expression in PBMCs in BRCA1 patients and patients in acute stage, while FOXO3a expression was significantly decreased in these patients. MiR-18a and miR-20a expression was not affected. We propose that expressional changes reported in this study could provide significant additive information for early BC diagnosis, disease development prediction and therapy outcome monitoring.

Tauroursodeoxycholic acid in the treatment of patients with amyotrophic lateral sclerosis.

BACKGROUND AND PURPOSE: Tauroursodeoxycholic acid (TUDCA) is a hydrophilic bile acid that is produced in the liver and used for treatment of chronic cholestatic liver diseases. Experimental studies suggest that TUDCA may have cytoprotective and anti-apoptotic action, with potential neuroprotective activity. A proof of principle approach was adopted to provide preliminary data regarding the efficacy and tolerability of TUDCA in a series of patients with amyotrophic lateral sclerosis (ALS). METHODS: As a proof of principle, using a double-blind placebo controlled design, 34 ALS patients under treatment with riluzole who were randomized to placebo or TUDCA (1 g twice daily for 54 weeks) were evaluated after a lead-in period of 3 months. The patients were examined every 6 weeks. The primary outcome was the proportion of responders [those subjects with improvement of at least 15% in the Amyotrophic Lateral Sclerosis Functional Rating Scale Revised (ALSFRS-R) slope during the treatment period compared to the lead-in phase]. Secondary outcomes included between-treatment comparison of ALSFRS-R at study end, comparison of the linear regression slopes for ALSFFRS-R mean scores and the occurrence of adverse events. RESULTS: Tauroursodeoxycholic acid was well tolerated; there were no between-group differences for adverse events. The proportion of responders was higher under TUDCA (87%) than under placebo (P = 0.021; 43%). At study end baseline-adjusted ALSFRS-R was significantly higher (P = 0.007) in TUDCA than in placebo groups. Comparison of the slopes of regression analysis showed slower progression in the TUDCA than in the placebo group (P < 0.01). CONCLUSIONS: This pilot study provides preliminary clinical data indicating that TUDCA is safe and may be effective in ALS.

The eye-tracking computer device for communication in amyotrophic lateral sclerosis.

OBJECTIVE: To explore the effectiveness of communication and the variables affecting the eye-tracking computer system (ETCS) utilization in patients with late-stage amyotrophic lateral sclerosis (ALS). METHODS: We performed a telephone survey on 30 patients with advanced non-demented ALS that were provisioned an ECTS device. Median age at interview was 55 years (IQR = 48-62), with a relatively high education (13 years, IQR = 8-13). A one-off interview was made and answers were later provided with the help of the caregiver. The interview included items about demographic and clinical variables affecting the daily ETCS utilization. RESULTS: The median time of ETCS device possession was 15 months (IQR = 9-20). The actual daily utilization was 300 min (IQR = 100-720), mainly for the communication with relatives/caregiver, internet surfing, e-mailing, and social networking. 23.3% of patients with ALS (n = 7) had a low daily ETCS utilization; most reported causes were eye-gaze tiredness and oculomotor dysfunction. CONCLUSIONS: Eye-tracking computer system is a valuable device for AAC in patients with ALS, and it can be operated with a good performance. The development of oculomotor impairment may limit its functional use.

Expression of vesicle-associated membrane-protein-associated protein B cleavage products in peripheral blood leukocytes and cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis.

BACKGROUND AND PURPOSE: Vesicle-associated membrane-protein-associated protein B (VAPB) is an endoplasmic reticulum (ER) resident protein participating in ER function, vesicle trafficking, calcium homeostasis and lipid transport. Its N-terminal domain, named MSP, is cleaved and secreted, serving as an extracellular ligand. VAPB mutations are linked to autosomal-dominant motor neuron diseases, including amyotrophic lateral sclerosis (ALS) type 8. An altered VAPB function is also suspected in sporadic ALS (SALS). METHODS: The expression pattern of VAPB cleavage and secreted products in the peripheral blood leukocytes (PBL) and cerebrospinal fluid (CSF) of SALS patients and neurological controls was assessed. PBL from healthy controls were also analyzed. Assays were carried out through western blotting, using an anti-VAPB (N-terminal) antibody. RESULTS: Two VAPB fragments containing the MSP domain (17 kDa and 14 kDa molecular sizes) were identified in PBL of SALS and controls, with no significant differences amongst groups. In CSF, only the 14 kDa VAPB MSP fragment was expressed and a corresponding VAPA fragment was not detected. The CSF VAPB fragment was absent in 58.7% of SALS patients, of whom 79.2% were bulbar onset (P = 0.001, bulbar versus spinal). CONCLUSIONS: The absence of the CSF VAPB MSP fragment from most bulbar-onset SALS patients suggests a specific alteration of brain-derived VAPB cleavage and secretion in this group of patients, and hints at a role of VAPB in the pathophysiology of this motor neuron disease.

The use of electrical impedance tomography to the differential diagnosis of pathological mammographic/sonographic findings.

Our study involved 870 eligible women with suspected pathological breast lesion discovered by mammography (MMG) or ultrasound examination (USG) which were recommended to pass histological examination to verify the diagnosis. All patients included in our study were divided into two age groups: the 1st group - patients older than 40 years (total of 724 patients) , the 2nd group - patients younger than 40 years (total of 146 patients). The purpose of our study was to analyze the possibilities of electrical impedance tomography (EIT) implementation to the differential diagnosis of pathologic lesions of the breast either solely, or in a combination with MMG/USG. The following results were obtained: in the 1st group the average specificity of MMG was 79.5%; when added EIT the average specificity decreased to 72.8%. The sensitivity of MMG, by contrast, increased from 87.8% when using it as an independent method to 94.5% when added EIT. In the 2nd group the average specificity of USG was 90.2%; when added EIT the average specificity decreased to 86.4%. Similarly, as in the 1st group the sensitivity of USG increased from 86.7% when using it as an independent method to 93.3% when added EIT.Analysis of false-negative results of electrical impedance tomography depending on the stage of the process has shown that as the earlier stage of the disease and as the smaller is the dimensions of the tumor, the higher is the number of false-negative results. In addition, we observed the dependence of the false-negative results of the tumor grade. The results of our study show that the use of EIT in addition to MMG/USG can improve the sensitivity of these methods and to increase the rate of early detection of breast cancer with minimal economic costs and highly qualified staff time expenditures.

Psychosocial support needs of Slovak breast cancer survivors one and three years after breast conserving surgery.

OBJECTIVE: To assess psychosocial support needs among Slovak breast cancer survivors treated by breast conserving surgery. METHODS: Patients responded the EORTC QLQ 30.3 questionnaire and the self-elaborated BCPN (Breast Cancer Psychosocial Needs) questionnaire one and three years after breast conserving surgery (BSC) and modified radical mastectomy (MRM). RESULTS: Assessment of psychosocial distress related to breast cancer treatment have shown a remarkable time-related increase in scores of emotional distress symptoms among breast cancer patients treated with breast-conserving surgery. Analysis of psychosocial support needs of these patients has shown, that a substantial part of breast cancer survivors are feeling inadequate beneficial psychosocial support from health care providers (informational support), social oncology workers, and psychologists (emotional support). Insufficient targeted psychosocial support involved individual as well as group intervention (provided by breast cancer patients surviving the disease for long time). CONCLUSION: Unmet psychosocial needs from health care providers has been perceived as a major factor harming patients ability to cope with the disease and to maintain their family, marital and social functioning (Tab. 2, Fig. 1, Ref. 22).

Utilisation of electrical impedance tomography in breast cancer diagnosis.

BACKGROUNDS: Breast cancer presents a serious medical and social problem worldwide. Early detection is key to effective breast cancer treatment. Therefore, scientists are consistently looking for new diagnostic techniques that would be more efficient, easy to use and safe for the patient. The main task of this study was to evaluate the feasibility of a novel low-cost non-invasive technique called electrical impedance tomography (EIT) and to determine whether EIT can qualitatively supplement the existing traditional imaging techniques in the process of breast cancer diagnostics. PATIENTS AND METHODS: Randomly selected patients with mammographic and/or sonographic abnormalities were involved into the study. In total, 808 patients aged 18-94 (mean 54) years participated in the survey. Exclusion criteria involved previous breast surgery, breast core biopsy or fine needle aspiration within the last 1 and 3 months, respectively. Furthermore, patients with implanted electrically powered devices (cardioverter, pacemaker) and patients previously treated by chemo-radiotherapy were also excluded. The EIT examination was performed using the electrical impedance computer mammograph MEIK developed by the Institute of Radio Engineering and Electronics, Russian Academy of Sciences. RESULTS: The following results were obtained: sensitivity of EIT was 87%, X-ray mammography (MMG) 89% and ultrasonography (USG) 91%; specificity of EIT was 85%, MMG 91% and USG 84%. Negative predictive value (NPV) of all three modalities showed nearly equal values, with slight advantage of the USG. MMG had the highest positive predictive (PPV) value (83%), EIT had the lowest (63%). Sensitivity increased to 96% and 98%, respectively, when combinations EIT+MMG and EIT+USG were used. The specificity increased to 79% for EIT+MMG and 71% for EIT+USG. EIT+MMG and EIT+USG NPV remained the same. PPV was 65% and 58%, respectively for the EIT+MMG and EIT+USG combination. CONCLUSION: Our study findings are comparable to those of other similar studies. Although the EIT is a promising method and deserves close attention of specialists, it cannot replace MMG and/or USG examination as it does not provide information on structural changes to the breast. It can, however, provide very useful additional information about metabolic processes in the body. But in order to show its full potential some improvements should be held.

Fatigue, sleep, and nocturnal complaints in patients with amyotrophic lateral sclerosis.

BACKGROUND AND PURPOSE: Fatigue is a common symptom in amyotrophic lateral sclerosis (ALS). Although sleep disturbances are a candidate factor that may interfere with fatigue in patients with ALS, the role of sleep-related abnormalities in determining fatigue in ALS is unknown. OBJECTIVE: To evaluate the frequency and determinants of fatigue in a group of 91 consecutive patients with ALS, with special attention to the relationship between fatigue and sleep problems. METHODS: Measures included the Fatigue Severity Scale (FSS), Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS), ALS Functional Rating Scale-Revised (ALSFRS-R), and Beck Depression Inventory (BDI). RESULTS: The mean FSS score was 4.35 ± 1.1, and 48 patients with ALS (52.75%) reported clinical significant fatigue. FSS score correlated with ALSFRS-R score, forced vital capacity, ESS, BDI, and global PSQI score. Patients with fatigue were significantly more disabled and more frequently reported difficulties staying asleep and nocturnal complaints, such as nocturia and disturbing muscle cramps. After multivariate analysis, patients' disability and nocturnal complaints were significantly associated with fatigue. CONCLUSION: In this study, we demonstrated that fatigue, a troublesome and disabling symptom in ALS, is associated with physical impairment and night-time complaints (such as nocturia and muscle cramps), suggesting that treating sleep problems might be useful in alleviating fatigue in these patients.

A simple and efficient method for DNA extraction from skin and paraffin-embedded tissues applicable to T-cell clonality assays.

PCR-based clonality assay of rearranged T-cell receptor genes gamma and beta (TCRG and TCRB) in a number of cases could be essential to discriminate between cutaneous T-cell lymphomas and reactive lymphoproliferative lesions in the skin. However, extraction of good-quality DNA from skin specimens (especially formalin-fixed paraffin-embedded) remains a challenge. Common procedures, being labour-intensive and time-consuming and requiring toxic solvents such as phenol and chloroform, still may end up with DNA sample of insufficient quality. We herewith present a simple and efficient method for DNA isolation based on ammonia extraction of tissue, followed by neutralization and simultaneous salting out of proteins with acetic acid. We have analysed 30 samples - 24 fresh (16 skin, two spleen and six lymph node) and six paraffin-embedded. Standard procedure (proteinase K digestion, followed by phenol/chloroform extraction) has been carried out simultaneously. We observed good PCR signal for TCRG rearrangements in 30 samples processed with the new protocol and only in 20 extracted with proteinase K/phenol/chloroform. For TCRB, the success rate was 29 of 30 with the new protocol, compared to 11 of 30 with conventional protocol. The proposed method of DNA extraction should improve the value of T-cell clonality assay, because insufficient DNA quality and quantity may bias analysis towards monoclonality and therefore cause false-positive results.