Development of New Predictive Equations for the Resting Metabolic Rate (RMR) of Women with Lipedema.

This study aimed to develop a novel predictive equation for calculating resting metabolic rate (RMR) in women with lipedema. We recruited 119 women diagnosed with lipedema from the Angiology Outpatient Clinic at Wroclaw Medical University, Poland. RMR was assessed using indirect calorimetry, while body composition and anthropometric measurements were conducted using standardized protocols. Due to multicollinearity among predictors, classical multiple regression was deemed inadequate for developing the new equation. Therefore, we employed machine learning techniques, utilizing principal component analysis (PCA) for dimensionality reduction and predictor selection. Regression models, including support vector regression (SVR), random forest regression (RFR), and k-nearest neighbor (kNN) were evaluated in Python's scikit-learn framework, with hyperparameter tuning via GridSearchCV. Model performance was assessed through mean absolute percentage error (MAPE) and cross-validation, complemented by Bland-Altman plots for method comparison. A novel equation incorporating body composition parameters was developed, addressing a gap in accurate RMR prediction methods. By incorporating measurements of body circumference and body composition parameters alongside traditional predictors, the model's accuracy was improved. The segmented regression model outperformed others, achieving an MAPE of 10.78%. The proposed predictive equation for RMR offers a practical tool for personalized treatment planning in patients with lipedema.

Disability and emotional symptoms in women with lipedema: A comparison with overweight/obese women.

BACKGROUND: Lipedema is characterized by the painful abnormal deposition of adipose tissue in the lower limbs and is often misdiagnosed as obesity. Considering the numerous bothersome physical symptoms of lipedema, women with lipedema may have greater disability and emotional problems than women with lifestyle-induced obesity. OBJECTIVES: Our study aims to assess disability, anxiety and depression symptoms in women with lipedema compared to women with overweight/obesity. MATERIAL AND METHODS: Women with lipedema (n = 45, with a mean age of 41 years) and women who are overweight/obese (n = 43, with a mean age of 44.95 years) were asked to complete the following questionnaires: The World Health Organization Disability Assessment Schedule (WHO-DAS II), Beck's Depression Inventory - II (BDI-II), and The Hospital Anxiety and Depression Scale (HADS). RESULTS: Despite the higher BMI in the overweight/obesity group, the group with lipedema was more disabled in numerous domains of the WHO-DAS II questionnaire, including Life activities - domestic, work and school responsibilities and Participation in society When the influence of BMI was adjusted, a difference in the domain of Mobility was also present. The study groups did not differ in anxiety and depression symptoms. CONCLUSIONS: We showed that behavioral impairment was the main factor affecting functioning in women with lipedema. Emotional symptoms did not differentiate the study groups. Leg volumes and adipose tissue pain intensity were associated with greater disability in women with lipedema, and should be considered in managing women with this condition and in future research estimating the effectiveness of lipedema treatment.

The Benefits of Low-Carbohydrate, High-Fat (LCHF) Diet on Body Composition, Leg Volume, and Pain in Women with Lipedema.

This study aimed to assess the potential benefits of a low-carbohydrate, high-fat (LCHF) diet on body composition, leg volume, and pain reduction in women with lipedema compared to overweight or women with obesity. The study included 113 female participants, 56 with lipedema and 57 with overweight/obesity (BMI >25 kg/m2) without lipedema. All subjects were prescribed a low-carbohydrate, high-fat (LCHF) diet with anti-inflammatory properties to adhere to for a duration of 7 months. Measurements of anthropometry, body weight, composition, and pain (VAS) were conducted at the study's commencement and conclusion. 52 participants completed the study. Both groups experienced a similar weight reduction, amounting to 12.9% compared to the baseline (-10.8 kg vs. -11.9 kg; p = 0.14, for lipedema and women with overweight/obesity, respectively). The most reduction was in body fat mass. Improvements in various parameters were observed, except for ankle circumferences, which decreased more in the lipedema group. Lipedema participants showed significantly reduced pain levels following the LCHF diet (4.6 ± 2.6 vs 3.0 ± 2.3; p < 0.001). The LCHF diet holds promise for weight loss, body fat reduction, leg volume management, and pain alleviation in women with lipedema. These findings provide valuable insights into potential therapeutic strategies for lipedema management.

The Effect of a Low-Carbohydrate High-Fat Diet on Laboratory Parameters in Women with Lipedema in Comparison to Overweight/Obese Women.

The aim of this study was to evaluate alterations in blood parameters after a low-carbohydrate high-fat (LCHF) diet in women with lipedema in comparison to overweight or obese women. A total of 115 women were classified into two groups: the lipedema group and the overweight/obesity group. Both study groups followed the caloric-restricted LCHF diet for 7 months. A total of 48 women completed the study. A reduction in body weight was observed in both study groups. A significant decrease in triglycerides and an increase in HDL-C concentrations were observed in both study groups. Despite the increase in the concentration of LDL-C observed in the lipedema group, changes in LDL-C differed between individual patients. Improvements in liver parameters, glucose tolerance, and a decrease in fasting insulin levels were observed, although they were less pronounced in the lipedema group than in the overweight/obesity group. Kidney and thyroid functions were similar before and after the LCHF diet in both groups. The LCHF diet may be a valuable nutritional strategy for lipedema and overweight/obese women, with a beneficial effect on weight, glucose profile, liver function, the concentration of triglycerides, and HDL-C and with no effect on kidney and thyroid function.

Rindera graeca (A. DC.) Boiss. & Heldr. (Boraginaceae) In Vitro Cultures Targeting Lithospermic Acid B and Rosmarinic Acid Production.

The in vitro cultures of Rindera graeca, a rare endemic plant, were developed as a sustainable source of phenolic acids. Various shoot and root cultures were established and scaled up in a sprinkle bioreactor. A multiplication rate of 7.2 shoots per explant was achieved. HPLC-PDA-ESI-HRMS analysis revealed the presence of rosmarinic acid (RA) and lithospermic acid B (LAB) as the main secondary metabolites in both the shoot and root cultures. The maximum RA (30.0 ± 3.2 mg/g DW) and LAB (49.3 ± 15.5 mg/g DW) yields were determined in root-regenerated shoots. The strongest free radical scavenging activity (87.4 ± 1.1%), according to 2,2-diphenyl-1-picrylhydrazyl-hydrate assay, was noted for roots cultivated in a DCR medium. The highest reducing power (2.3 µM ± 0.4 TE/g DW), determined by the ferric-reducing antioxidant power assay, was noted for shoots cultivated on an SH medium containing 0.5 mg/L 6-benzylaminopurine. A genetic analysis performed using random amplified polymorphic DNA and start codon targeted markers revealed genetic variation of 62.8% to 96.5% among the investigated shoots and roots. This variability reflects the capacity of cultivated shoots and roots to produce phenolic compounds.

Lymphoscintigraphic alterations in lower limbs in women with lipedema in comparison to women with overweight/obesity.

Introduction: Lipedema is a bilateral enlargement of the legs due to abnormal depositions of subcutaneous fat. Recent studies using lymphoscintigraphy documented that lipedema associates with lymphatic alterations. It is still not known, whether non-lipedema obesity also leads to similar lymphoscintigraphic changes within lower legs. Clinically, both, lipedema and obesity may progress to secondary lymphedema. The aim of the study was to evaluate lymphoscintigraphy of lower limbs in women with lipedema in comparison to overweight/obese women. Methods: 51 women (in the mean age of 43.3 ± 13.56) with the diagnosis of lipedema and 31 women (in the mean age of 44.7 ± 13.48) with overweight/obesity were enrolled into the study. Women in both study groups had no clinical signs of lymphedema. The groups were matched by mean volume of their legs, calculated using the formula for a truncated cone. Lymphoscintigraphy was evaluated in every women qualitatively. Body composition parameters were assessed using bioelectric impedance analysis (BIA). Results: Lymphoscintigraphic alterations within lower extremities were similar in both, lipedema and overweight/obese groups and were present in majority of women in both study groups. The most common lymphoscintigraphic alteration in both groups were additional lymphatic vessels (in the lipedema group observed in 76.5% of patients and in the overweight/obesity group - in 93.5%). Visualization of popliteal lymph nodes and dermal backflow were observed respectively in 33% and in 5.9% in the group with lipedema and in 45.2% and in 9.7% in the overweight/obesity group. There were significant relationships between severity of lymphoscintigraphic alterations and weight, lean body mass (LBM), total body water (TBW), volume of both legs and thigh circumference in the lipedema group. Such relationships were absent in the overweight/obesity group. Discussion: Our study indicates that lymphatic alterations are present before development to clinically visible secondary lymphedema in both conditions, lipedema and overweight/obesity. In majority of women from both study groups they indicate rather an overload of the lymphatic system than insufficiency. Lymphoscintigraphic alterations are similar in both groups, therefore, lymphoscintigraphy is not a diagnostic tool that might distinguish lipedema from overweight/obesity.

The Role of Flaxseed in Improving Human Health.

Flaxseed contains high amounts of biologically active components such as α-linolenic acid, lignans, and dietary fiber. Due to its numerous nutritional properties, flaxseed has been classified as a "superfood", that is, a food of natural origin with various bioactive components and many health-promoting benefits. Flaxseed consumption can be an important factor in the prevention of diseases, particularly related to nutrition. The regular consumption of flaxseed may help to improve lipid profile and lower blood pressure, fasting glucose, and insulin resistance index (HOMA-IR). Moreover, flaxseed is characterized by anticancer and antioxidant properties and can significantly reduce the intensity of symptoms associated with menopause, constipation, and mental fatigue, improve skin condition, and accelerate wound healing. In addition to its bioactive compounds, flaxseed also contains antinutrients such as cyanogenic glycosides (CGs), cadmium, trypsin inhibitors, and phytic acid that can reduce the bioavailability of essential nutrients and/or limit its health-promoting effects. Three common forms of flaxseed available for human consumption include whole flaxseed, ground flaxseed, and flaxseed oil. The bioavailability of ALA and lignans is also dependent on the form of flaxseed consumed. To ensure high bioavailability of its bioactive components, flaxseed should be consumed in the ground form.

Comparison of Actual and Predicted Resting Metabolic Rate in Women with Lipedema.

Background: An adequate dietary energy supply is particularly important in patients with lipedema as it promotes weight and fat loss. Accurate estimation of resting metabolic rate (RMR) allows implementing a proper calorie restriction diet in patients with lipedema. Our study aimed to compare actual resting metabolic rate (aRMR) with predicted resting metabolic rate (pRMR) in women with lipedema and to determine the association between individual body composition parameters, body mass index, and aRMR. Methods and Results: A total of 108 women diagnosed with lipedema were enrolled in the study. aRMR was obtained by indirect calorimetry (IC) using FitMate WM metabolic system (Cosmed, Rome, Italy). pRMR was estimated with predictive equations and bioelectric impedance analysis (BIA). All body composition parameters were based on BIA. The mean aRMR in the study group was 1705.2 ± 320.7 kcal/day. This study found the agreement of predictive equations compared to IC is low (<60%). Most methods of predicted RMR measurement used in our study significantly underpredicted aRMR in patients with lipedema. Therefore, the most applied equations remain useless in clinical practice in this specific population due to large individual differences among the studied women. Conclusions: IC is the best tool to evaluate RMR in evaluated patients with lipedema. It is necessary to propose a new equation to RMR determination in clinical practice.

The Effect of a Low-Carbohydrate, High-Fat Diet versus Moderate-Carbohydrate and Fat Diet on Body Composition in Patients with Lipedema.

Introduction: Lipedema is a disorder characterized by an excessive accumulation of subcutaneous body fat, mainly bilateral and symmetrical accumulation of fat deposits, particularly in the lower extremities excluding feet. Pain (spontaneous or with palpation) and increased capillary fragility with bruising are also part of clinical presentation. It is estimated to occur in approximately 11.0% of women worldwide. Management of obesity among patients with lipedema is a key component in its treatment. Purpose: The aim of this study was to compare effectiveness of two diets: low-carbohydrate-high-fat diet (LCHF) and medium-fat-medium-carbohydrate diet (MFMC) in body weight, body fat and limb circumference reduction in patients with lipedema. Material and Methods: The studied women (n = 91) were divided into 2 groups and submitted to 1 of the 2 diets for 16 weeks. Anthropometric measurements such as body height [cm], body weight [kg], body fat percentage [%], body fat [kg], lean body mass [kg], and visceral fat level were collected at the beginning and end of the study. Results: We have not found any significant differences in anthropometric measurements at the baseline between groups. Body weight and all anthropometric parameters decreased significantly in both groups after 16 weeks of diets, excluding the circumference above the right ankle for the MFMC diet which did not change. The LCHF diet contributed to reduction of body weight (-8.2 ± 4.1 kg vs -2.1 ± 1.0 kg; p < 0.0001), body fat (-6.4 ± 3.2 kg vs 1.6 ± 0.8 kg; p < 0.0001), waist (-7.8 ± 3.9 cm vs -2.3 ± 1.1 cm; p < 0.0001), hips (-7.4 ± 3.7 cm vs -2.5 ± 1.3 cm; p < 0.0001), thighs and calves' circumferences compared with the MFMC diet. We observed reduction of pain in the extremities and mobility improvement in LCHF group (data not shown). Conclusion: The LCHF diet was more effective than MFMC in body weight, body fat and lower limb circumferences reduction.

Mediterranean diet adherence among cosmetology students.

BACKGROUND: The Mediterranean diet (MD) is universally recognized as the healthiest model of nutrition whose beneficial effects help prevent many diet-related diseases. OBJECTIVE: The aim of the study was to assess cosmetology students' adherence to the Mediterranean Diet using the Mediterranean Diet Score (MDS). MATERIAL AND METHODS: The study group consisted of 175 cosmetology students of the School of Physiotherapy whose dietary habits were assessed using a validated food frequency questionnaire (FFQ) consisting of 154 food and drink items representative of the diet during the last year. We analyzed data obtained from FFQ and calculated the average number of daily and/or weekly servings from each of the 9 MDS food groups (grains, vegetables, fruits/nuts, milk/dairy products, meat/meat products, legumes, alcohol, olive oil, fish). To obtain more reliable results, we analyzed other products consumed by study participants (sweets, beverages, eggs and potatoes). Study participants were assigned 0, 1 or 2 points for each MDS ingredient. Mediterranean diet adherence was assessed on a 17-point scale. Low adherence to MD was defined at 0-7 points, moderate at 8-10 points, and high at 11-17 points. RESULTS: The mean MDS was 7.1 ± 2.3. Students who were assigned 11-17 points (n = 20) consumed significantly more vegetables, fruits, nuts, legumes, fish, olive oil and significantly less meat, meat products, milk, dairy, and sweets compared to study participants who were assigned the lowest number of points (n = 78). We reported significant differences in the amounts of consumed vegetables and sweets between participants who received 8-10 points (n = 77) and those who received 11-17 points. The higher MDS was significantly associated with the higher intakes of vegetables and dietary fiber. CONCLUSIONS: The dietary patterns of study group of Polish cosmetology students did not adhere to the MD recommendations.

Chemical Profile and Screening of Bioactive Metabolites of Rindera graeca (A. DC.) Bois. & Heldr. (Boraginaceae) In Vitro Cultures.

Rindera graeca is a rare endemic plant where in vitro culture has been used in order to investigate bioactive metabolites. Phytochemical study of the in vitro shoots and hairy roots led to the isolation of seven phenolic derivatives and the unusual furano-naphthoquinone rinderol. R. graeca was also analyzed for its pyrrolizidine alkaloids content by LC-MS, and it was found to contain echinatine together with echinatine and rinderine N-oxides. Rinderol, isolated only from in vitro hairy root culture for the first time in the genus, revealed promising bioactivities. It was evaluated in vitro against a panel of microorganisms, showing very strong activity specifically against Gram-positive bacteria (MIC values 0.98 × 10-2-1.18 µg/mL) as well as very interesting antiproliferative effect against the human non-small-cell bronchopulmonary carcinoma cell line NSCLC-N6-L16 and the epidermoid lung cancer cell line A549. These findings were compared with the chemical profile of the plant from nature, while this study is the first to report on the effects of R. graeca extracts obtained from in vitro culture, providing a valuable contribution to the scientific community towards this sustainable method of production of potential bioactive molecules.

The role of nutrition in Alzheimer's disease.

The aging population is a significant social, medical and economic problem due to increasing prevalence of chronic diseases in elderly population. Alzheimer's disease (AD) is the most common form of dementia and the most common neurodegenerative disease. It is characterized by a progressive deterioration of memory and cognitive function. So far, there is neither an effective prevention nor cure for dementia, so more and more attention is paid to the prevention of this group of diseases, particularly to the appropriate diet. Preventive intervention gives the best results if introduced before the first symptoms of dementia, i.e., around the age of 50. This is when the nutritional status, number of synapses, cognition, and neuropathological changes in the nervous system compensate each other, which increases the chances of staying healthy for a longer period of time. It has been proven that dietary habits, which lead to the development of cardiovascular and metabolic diseases, significantly increase the risk of dementia. On the other hand, a Mediterranean diet rich in antioxidants, fiber and omega-3 polyunsaturated fatty acids may have a protective effect on the neurodegenerative process. The beneficial effect of many nutrients on the course of AD has been demonstrated. These include: glutathione, polyphenols, curcumin, coenzyme Q10, vitamins B6, B12, folic acid, unsaturated fatty acids, lecithin, UA, caffeine and some probiotic bacteria. A diet rich in saturated fatty acids and branched-chain amino acids (BCAA) promotes the progression of dementia. Dietary intervention should be introduced as early as possible to minimize the risk of developing dementia. The Mediterranean and DASH diets have been documented to protect against AD. However, the MIND diet is reported to be much more effective in preventing cognitive decline/dementia than either the Mediterranean or DASH diets alone.

Polyurethane Foam Rafts Supported In Vitro Cultures of Rindera graeca Roots for Enhanced Production of Rinderol, Potent Proapoptotic Naphthoquinone Compound.

Unique phytochemical profile of plants belonging to Boraginaceae family provides a prolific resource of lipophilic pigments from the group of naphthoquinone derivatives. To overcome low compound content, the major obstacle of plant-based production, immobilization of Rindera graeca roots in in vitro cultures was implemented for efficient production of rinderol, novel furanonaphthoquinone derivative with anticancer properties. Chromatographic procedures revealed rinderol presence in extracts of all investigated root lines, derived both from root biomass and post-culture medium. Unexpectedly, in the second stage of the experiment, rinderol production was ceased in control, unmodified culture systems. On the contrary, roots immobilized on PUF rafts uniformly and stably produced rinderol, and its highest amount was noted for transformed root lines after 42 days of cultivation (222.98 ± 10.47 µg/flask). PUF occurred to be the main place of compound accumulation. Moreover, investigation of rinderol biological activity revealed its fast-acting cell death induction in HeLa cervical cancer cells at relatively low concentrations. Presented results revealed successful application of R. graeca roots immobilization on PUF rafts for production and in situ product removal of rinderol, novel lipophilic furanonaphthoquinone with suggested proapoptotic activity.

Assessment of eating and lifestyle habits among Polish cosmetology and physiotherapy students.

BACKGROUND: Proper nutritional behaviors are one of the most important factors shaping our health, as they influence people's physical and mental state. Good eating habits help prevent many chronic diseases, including cardiovascular disease, type II diabetes, some types of cancer, and osteoporosis. The Polish population aged 20-34 years displays improper nutritional behaviors which may put them at risk of developing diet-dependent diseases in the future. There is a need to assess students' lifestyle, including their eating habits, as it would help adapt already existing education programs. OBJECTIVE: We evaluated and compared eating habits and lifestyle of students studying cosmetology and physiotherapy in association with selected risk factors of civilization diseases. MATERIAL AND METHODS: We used a standardized dietary questionnaire evaluating respondents' physical activity, smoking and dietary habits. The obtained results were compared with the recommendations from the Polish food-based dietary guidelines. We evaluated respondents' nutritional status based on their anthropometric measurements such as body weight, body height, hips and waist circumferences and then we estimated their body mass index (BMI) and waist hip ratio (WHR). RESULTS: Students studying cosmetology and physiotherapy obtained on average 5.3 ±1.5 points based on their compliance with the nutritional recommendations. The academic youth did not meet principles of healthy nutrition according to the given recommendations. There were no significant differences in nutrition status between the faculties, except for products that provide complete protein. CONCLUSIONS: There is a need to increase nutritional awareness of academic youth, especially students of health-related faculties, because better knowledge on nutrition would help them choose healthier food options.

The influence of diet on gastrointestinal Candida spp. colonization and the susceptibility of Candida spp. to antifungal drugs

Background: Candida spp. has been identified as the most common member of human gut microbiota. This yeast-like fungus is recognized as an opportunistic organism due to its potential to cause diseases in all parts of the gastrointestinal tract. Objective: The aim of the study was to investigate the relationship between diet and health of the study participants and the presence of gastrointestinal Candida spp. Susceptibility of Candida to antifungal drugs was determined. Material and methods: Material comprised of oral swabs and fecal samples self-collected by the study participants. The participants filled in a nutritional questionnaire. A total of 30 people took part in the study, including 28 women and 2 men. The study was conducted in Wroclaw, Poland. Susceptibility of Candida to antifungal drugs was determined using Bio-Rad's FUNGITEST™ Kit designed for the susceptibility testing of yeasts to six antifungal drugs. Results: In the group with negative Candida spp. cultures, healthier wheat flour substitutes were consumed significantly more frequently than in the group with positive Candida spp. cultures. Yellow cheeses and quark were eaten significantly more frequently in the group with negative Candida spp. cultures. No antifungal resistance was detected in the study group. Conclusions: The increased consumption of purified wheat flour products was linked to the presence of gastrointestinal Candida spp. A higher consumption of cheese was observed in the group with negative Candida spp. cultures, which may indicate the inhibitory effect of saturated fatty acids on the growth of human Candida spp. Sensitivity of C. albicans to antifungal drugs may increase effectiveness of candidiasis treatment.

Hairy Root Cultures for the Production of Anti-cancer Naphthoquinone Compounds.

BACKGROUND: Recent years have brought the dynamic development in studies of naphthoquinones obtained from plants, in vitro cultures and semi- or total synthesis. This review presents the hairy root cultures approach for producing naphthoquinones and summarizes their most recent anti-cancer investigations. OBJECTIVE: This review aimed to define biotechnological strategies impacted on naphthoquinones production in hairy root cultures. Up to now the major source of shikonin/alkannin derivatives, rhinacanthins and ramentaceone is isolation from plant material, also derived via biotechnological methods. Moreover, the most recent anti-cancer activity studies on naphthoquinones which could be produced in hairy root cultures were outlined. METHODS: For databases survey two selection criteria were used: (i) naphthoquinone could be produced in hairy roots, and (ii) it exhibits anti-cancer properties. RESULTS: Ninety two papers were included in the review, thirty described biotechnological approaches enhancing naphthoquinones production, among them twenty seven were dedicated to hairy root cultures. Forty papers outlined the anti-cancer activity of targeted naphthoquinones including the type of cancer and bioassays description. The synergistic effect of natural naphthoquinones and other anti-cancer therapies was reviewed and toxicity of natural naphthoquinones and plant extracts was discussed. The review highlights tendencies in hairy root investigations and indicates the possible future research directions for improving biotechnological production efficacy. CONCLUSION: This review demonstrates a great potential of hairy root cultures for naphthoquinones production, which could be furtherly developed for future medical purposes, especially as anti-cancer agents. This area of plant biotechnology will be surely still developed with traditional and new strategies.

The chemical profile of pyrrolizidine alkaloids from selected greek endemic boraginaceae plants determined by gas chromatography/mass spectrometry.

Four Greek endemic Boraginaceae plants, Onosma erecta Sibth. & Sm., Onosma kaheirei Teppner, Onosma leptantha Heldr., and Cynoglossum columnae L. (aerial parts), were screened for their content of pyrrolizidine alkaloids (PAs). TLC with the Mattocks-Molyneux visualization reagent was used as a preliminary qualitative test for PA or PA N-oxide detection. The extracts of the species found to contain PAs and their N-oxides were further analyzed by GC/MS, so as to identify their structures by means of the mass spectra and retention index values of known PAs already published in the literature. Twenty-three PAs were identified. For additional peaks, recognized as possible PAs by their MS pattern, no exact structures were tentatively suggested, as a result of lack of matching literature data. Furthermore, a quantitative PA profile of the species was obtained.

Effect of l-phenylalanine on PAL activity and production of naphthoquinone pigments in suspension cultures of Arnebia euchroma (Royle) Johnst.

The effects of l-phenylalanine (PHE) on cell growth and production of shikonin and its derivatives, acetylshikonin (ACS) and isobutyrylshikonin (IBS), in suspension cultures of Arnebia euchroma were examined. Supplementing media using PHE have been successfully utilized to enhance shikonin production in cell cultures of other species of Boraginaceae. l-Phenylalanine, the key compound in the phenylpropanoid pathway, is converted by phenylalanine ammonia lyase (PAL) to trans-cinnamic acid, which is the precursor of p-hydroxybenzoic acid (PHB). Coupling of PHB and geranyl pyrophosphate (derived from mevalonate pathway) by p-hydroxybenzoate-m-geranyltransferase leads later to biosynthesis of shikonins. The addition of 0.01 or 0.1 mM PHE to the culture medium stimulated cell proliferation, where the highest observed increase in fresh cell biomass (measured as a ratio of final weight to initial weight) was 12-fold, in contrast to an eightfold increase in control cultures. Whereas, growth media supplemented with 1 mM PHE markedly reduced the rate of cell growth (to only twofold). Precursor feeding had detrimental effects on both ACS and IBS production in all PHE-supplemented media. The highest total content (intracellular + extracellular) of the investigated red pigments (9.5 mg per flask) was detected in the control culture without PHE. ACS was the major component of the naphthoquinone fraction determined in cells and post-culture media. Shikonin itself was found only in the post-culture media from cultures supplemented with 0.01 or 0.1 mM PHE. Increases in PAL activity corresponded well with the accumulation of investigated naphthoquinones in control culture. However, peak PAL activity did not directly correlate with maximum production of shikonin derivatives. Cytotoxicity of extracts, prepared from the cells cultivated in the presence of PHE or in control cultures, was tested on three cancer cell lines: HL-60, HeLa, and MCF-7. The extracts prepared from the untreated control cultures proved to be the most potent against the examined cancer cell lines. The mean inhibitory concentration values were 0.3, 13, and 8 µg ml(-1) for the HL-60, HeLa, and MCF-7 cells, respectively.