Economic burden of clinical trials in lung cancer in a German Comprehensive Cancer Center.

OBJECTIVES: The recent success of individualized lung cancer therapy has triggered fundamental changes in clinical research strategies. To date there is a strong focus on early proof of concept trials in genetically preselected small patient subgroups. This analysis focuses on the economic burden caused by such trials for advanced lung cancer patients in a German Comprehensive Cancer Center (CCC). METHODS: The profit margins between recruiting groups with ≤3 and >3 patients were compared. Clinical and economic data from clinical trials for advanced lung cancer (LC), pharma-sponsored trials (PhST) as well as investigator initiated trials (IIT), conducted between 2011 and 2015 at the Center for Integrated Oncology (CIO) Cologne, were analyzed using a profit-center calculation model. RESULTS: 161 patients were enrolled in 27 clinical trials. The key economic parameter determining costs and payments was the 'trial visits'. In comparison of the two groups (A≤3; B>3 patients enrolled) we found negative profit margins for the low recruiting group (€ -1444). Concerning the number of visits significant differences were found between PhST and IIT (p=0.009). Additionally, sub-analysis show structural differences in cost composition by conducting PhST and IIT. CONCLUSION: Trials with low patient numbers and IIT, do not cover the cost. To ensure adequate, cost-covering compensation by pharmaceutical companies CCCs have to thoroughly calculate the cost of early proof of concept trials. The findings of this study also underline the need for novel structures in public funding for investigator-initiated clinical trials in precision medicine.

Memory and multitasking performance during acute allergic inflammation in seasonal allergic rhinitis.

BACKGROUND: In previous research, patients with seasonal allergic rhinitis (SAR) showed poorer school and work performance during periods of acute allergic inflammation, supporting the idea of an impact of SAR on cognitive functions. However, the specific cognitive domains particularly vulnerable to inflammatory processes are unclear. OBJECTIVE: In this study, the influence of SAR on memory and multitasking performance, as two potentially vulnerable cognitive domains essential in everyday life functioning, was investigated in patients with SAR. METHODS: Non-medicated patients with SAR (n = 41) and healthy non-allergic controls (n = 42) performed a dual-task paradigm and a verbal learning and memory test during and out of symptomatic allergy periods (pollen vs. non-pollen season). Disease-related factors (e.g. symptom severity, duration of symptoms, duration of disease) and allergy-related quality of life were evaluated as potential influences of cognitive performance. RESULTS: During the symptomatic allergy period, patients showed (1) poorer performance in word list-based learning (P = 0.028) and (2) a general slowing in processing speed (P < 0.001) and a shift in processing strategy (P < 0.001) in multitasking. Yet, typical parameters indicating specific multitasking costs were not affected. A significant negative association was found between learning performance and duration of disease (r = -0.451, P = 0.004), whereas symptom severity (r = 0.326; P = 0.037) and quality of life (r = 0.379; P = 0.015) were positively associated with multitasking strategy. CONCLUSIONS: Our findings suggest that SAR has a differentiated and complex impact on cognitive functions, which should be considered in the management of SAR symptoms. They also call attention to the importance of selecting sensitive measures and carefully interpreting cognitive outcomes.

Body composition, tissue deposition, and lysine utilization for protein deposition of barrows and gilts fed crystalline or protein-bound lysine.

An experiment with 2 trials (28 d/trial) was conducted to determine body composition, tissue deposition, and utilization of Lys for protein deposition (PD) of barrows and gilts fed -Lys·HCl (CLys) or protein-bound Lys in soybean meal (SBM). Thirty-two growing pigs (16 barrows and 16 gilts; average initial BW of 18.6 kg) were used in each of 2 trials. Four pigs (2 barrows and 2 gilts) were euthanized at the start of each trial to determine initial body composition. The remaining pigs were euthanized at the end of the trials to determine empty-body composition and deposition rates of water, protein, fat, ash, and AA. Pigs were randomly allotted to 1 of 7 dietary treatments. There were 2 replications per treatment in each trial for a total of 4 replications. Dietary treatments consisted of a corn-SBM basal diet (0.48% Lys) and diets containing 0.56%, 0.65%, and 0.74% standardized ileal digestible (SID) Lys that were achieved by adding Lys to the basal diet from either SBM or CLys. Pigs fed the CLys-supplemented diets at 0.65% SID Lys had more ( < 0.05) body water (663 vs. 624 g/kg) and less ( < 0.01) body protein (153 vs. 160 g/kg) than pigs fed the SBM-supplemented diets. Body fat content decreased ( < 0.01) as the dietary Lys increased similarly for pigs fed Lys from SBM and pigs fed CLys. Gilts had greater ( = 0.05) body Lys content in body protein than barrows (7.68 vs. 7.52 g/100 g). Empty-body ash contents were not different between pigs fed CLys or SBM-supplemented diets. Water deposition and PD increased linearly ( < 0.01) with dietary Lys and were least ( < 0.01) in pigs fed the basal diet but were similar when comparing pigs fed CLys and SBM-supplemented diets at the same dietary Lys concentration. Lysine deposition showed a linear increase ( < 0.01) with dietary Lys but was not different between pigs fed the 2 Lys sources at the same concentration. Barrows and gilts did not differ in tissue deposition rates. Overall, empty-body contents and deposition rates of essential and nonessential AA were not different between pigs fed CLys and pigs fed SBM-bound Lys. The amount of SID Lys required for PD ranged between 0.09 and 0.13 g/g for both sources of Lys. The Lys deposition:SID Lys intake ratio was greater ( < 0.01) in gilts than barrows (0.62 vs. 0.56). Body composition, tissue deposition, and utilization of Lys for PD and Lys deposition were not different in pigs fed diets supplemented with -Lys·HCl with respect to protein-bound Lys in SBM.

Extraction and downstream processing of plant-derived recombinant proteins.

Plants offer the tantalizing prospect of low-cost automated manufacturing processes for biopharmaceutical proteins, but several challenges must be addressed before such goals are realized and the most significant hurdles are found during downstream processing (DSP). In contrast to the standardized microbial and mammalian cell platforms embraced by the biopharmaceutical industry, there are many different plant-based expression systems vying for attention, and those with the greatest potential to provide inexpensive biopharmaceuticals are also the ones with the most significant drawbacks in terms of DSP. This is because the most scalable plant systems are based on the expression of intracellular proteins in whole plants. The plant tissue must therefore be disrupted to extract the product, challenging the initial DSP steps with an unusually high load of both particulate and soluble contaminants. DSP platform technologies can accelerate and simplify process development, including centrifugation, filtration, flocculation, and integrated methods that combine solid-liquid separation, purification and concentration, such as aqueous two-phase separation systems. Protein tags can also facilitate these DSP steps, but they are difficult to transfer to a commercial environment and more generic, flexible and scalable strategies to separate target and host cell proteins are preferable, such as membrane technologies and heat/pH precipitation. In this context, clarified plant extracts behave similarly to the feed stream from microbes or mammalian cells and the corresponding purification methods can be applied, as long as they are adapted for plant-specific soluble contaminants such as the superabundant protein RuBisCO. Plant-derived pharmaceutical proteins cannot yet compete directly with established platforms but they are beginning to penetrate niche markets that allow the beneficial properties of plants to be exploited, such as the ability to produce 'biobetters' with tailored glycans, the ability to scale up production rapidly for emergency responses and the ability to produce commodity recombinant proteins on an agricultural scale.

Additional effects of homeopathy on chronic periodontitis: a 1-year follow-up randomized clinical trial.

BACKGROUND AND OBJECTIVE: The aim of this study was test the hypothesis that homeopathy (H) enhances the effects of scaling and root planing (SRP) in patients with chronic periodontitis (CP). MATERIALS AND METHODS: 50 patients with CP were randomly allocated to one of two treatment groups: SRP (C-G) or SRP + H (H-G). Assessments were made at baseline and after 3 and 12 months of treatments. The local and systemic responses to the treatments were evaluated by clinical and serologic parameters, respectively. RESULTS: Both groups displayed significant improvements, however, using clinical attachment gain and reductions in HDL, LDL and Total Cholesterol, Triglycerides, Glucose and Uric acid, from baseline to 1 year, as criteria for treatment success, H-G performed significantly better than C-G. CONCLUSION: The findings of this 1-year follow-up randomized clinical trial suggest that homeopathic medicines, as an adjunctive to SRP, can provide significant local and systemic improvements for CP patients.

[Acute mountain sickness : How can it be treated and how can it be avoided?]. / Höhenkrankheit : Wie behandeln, wie vermeiden?

Due to the decreasing partial pressure of oxygen, high altitude sickness can occur at heights over 2,500 m. This can be best avoided by slow adaptation to the altitude (acclimatization). In this way the organism adapts to the chronic hyperventilation and in the further process the oxygen content is normalized by an increase in erythrocytes. The commonest form of high altitude sickness is acute mountain sickness which is characterized by the leading symptom of headache. When additional signs of ataxia occur there is an acute danger of edema which is associated with a high mortality. Stress dyspnea, coughing and rasping breathing noises also occur by the potentially fatal high altitude pulmonary edema. All forms of high altitude sickness can be countered by a rapid descent to a height of at least 500 m. In acute mountain sickness acetazolamide can be administered (2 × 250 mg), for high altitude cerebral edema dexamethasone (3 × 4-8 mg) and for high altitude pulmonary edema nifedipine (initially 10 mg then 20 mg retard).

The impact of non-surgical periodontal treatment on serum levels of long chain-polyunsaturated fatty acids: a pilot randomized clinical trial.

BACKGROUND AND OBJECTIVE: Our group recently found higher levels of serum long chain-polyunsaturated fatty acids (LC-PUFAs) in patients with chronic periodontitis compared to controls. However, the effect of periodontal treatment on LC-PUFA serum levels has not been investigated. The primary aim of the present study was to investigate the impact of periodontal treatment on LC-PUFA serum levels. A secondary aim was to assess the effect of dietary ω-3 supplementation on clinical outcome. MATERIAL AND METHODS: The test group was composed of 10 patients with generalized chronic periodontitis (mean age 44 ± 6.4 years) treated with scaling and root planing associated with 4 mo of ω-3 supplementation eicosapetaenoic acid (EPA) plus docosahexaenoic acid (DHA), 3 g/d. The placebo group was composed of 11 patients (47.9 ± 10.5 years) that received scaling and root planing plus placebo. The periodontal examination included probing depth, clinical attachment level, bleeding on probing and visible plaque index. Docosapentaenoic acid (DPA), EPA, DHA and arachidonic acid (AA) were detected using gas chromatograph. RESULTS: In the placebo group, all LC-PUFAs levels reduced significantly (DHA, DPA and AA, p = 0.004; EPA, p = 0.008). In the test group, only DPA and AA showed a significant reduction (p = 0.005). Moreover, a significant decrease in the ratios AA/EPA and AA/DHA (p = 0.005) was observed in the test group. CONCLUSION: Non-surgical periodontal treatment reduced significantly the serum levels of all analyzed LC-PUFAs except those presented in the supplementation. The ω-3 dietary supplementation had no effect on clinical outcome of treatment.

High concentration of residual aluminum oxide on titanium surface inhibits extracellular matrix mineralization.

In the present study we characterized titanium (Ti) surfaces submitted to different treatments and evaluated the response of osteoblasts derived from human alveolar bone to these surfaces. Five different surfaces were evaluated: ground (G), ground and chemical etched (G1-HF for 60 s), sand blasted (SB-Al(2)O(3) particles 65 mum), sand blasted and chemical etched (SLA1-HF for 60 s and SLA2-HF for 13 s). Surface morphology was evaluated under SEM and roughness parameters by contact scanning instrument. The presence of Al(2)O(3) was detected by EDS and the amount calculated by digital analyses. Osteoblasts were cultured on these surfaces and it was evaluated: cell adhesion, proliferation, and viability, alkaline phosphatase activity, total protein content, and matrix mineralization formation. Physical and chemical treatments produced very different surface morphologies. Al(2)O(3) residues were detected on SB and SLA2 surfaces. Only matrix mineralization formation was affected by different surface treatments, being increased on rough surface (SLA1) and reduced on surface with high amount of Al(2)O(3) residues (SB). On the basis of these findings, it is possible to conclude that high concentration of residual Al(2)O(3) negatively interfere with the process of matrix mineralization formation in contact with Ti implant surfaces.

Lung disease severity, chronic inflammation, iron deficiency, and erythropoietin response in adults with cystic fibrosis.

Chronic lung disorders are usually associated with a hypoxia driven increase in red cell mass. However, patients with cystic fibrosis (CF) often have normal or decreased haemoglobin levels. The present prospective observational study in cystic fibrosis patients was performed to determine which factors were involved in alterations in the hematopoetic response to corresponding arterial oxygen pressure. Sixty adult patients (age 21-51) with stable CF were included. They all had vitamin A, D, E, and K but no vitamin B12 supplementation. Twenty-five patients were on oral Fe(2+) (100 mg/day). Resting arterial blood gases, lung function, complete blood counts, parameters of iron status, CRP, sputum microbiology and serum erythropoietin were measured at recruitment and after 3 and 6 months. Patients had varying degrees of pulmonary functional impairment and 9% were hypoxemic (arterial oxygen pressure <60 mm Hg). Low-grade systemic inflammation (CRP > 0.5 mg/dl) was present in 40% of the patients, who all had bacterial colonization. None of the patient had erythrocytosis and 12 patients had anemia. There was no significant difference in iron status between patients with or without chronic iron supplementation and erythropoietin levels were normal. During the 6 months observation period no significant changes occurred. The patients exhibited an impaired erythropoietic response to hypoxemia with normal or low hematocrit in spite of chronic lung disease which might be caused by chronic inflammation associated with CF. Linear multivariate regression analysis revealed CRP levels but neither iron substitution, nor erythropoietin levels nor lung function parameters as independent determinant of haemoglobin levels. CF may be associated with anemia of variable severity as expression of the chronic inflammation present in these patients. The therapeutic consequences are to treat the underlying inflammation rather than to supplement iron.

Genetic stability of recombinant potato virus X virus vectors presenting foreign epitopes.

We investigated the genetic stability of recombinant potato virus X vectors presenting beet necrotic yellow vein virus (BNYVV) epitopes. Following N-terminal PVX coat protein (CP) fusion of the BNYVV epitopes, we inoculated Nicotiana benthamiana plants with recombinant (r)PVX and carried out five serial passages through systemically-infected plants. RT-PCR investigation of the BNYVV epitope sequences revealed the accumulation of several point mutations and deletions, predominantly affecting positively-charged residues. A comparison of the isoelectric point (pI) values and charges of the wild type and rCPs showed that the initial high rCP pI values had changed to values closer to that of the wild-type CP.

Growth, carcass traits, and plasma amino acid concentrations of gilts fed low-protein diets supplemented with amino acids including histidine, isoleucine, and valine.

Three experiments were conducted to determine the fifth-limiting amino acid for growing pigs in an 11% CP, corn-soybean meal diet. In each experiment, 36 gilts (initial weight 19.5, 21.9, and 21.0 kg, respectively) were penned individually and fed one of six diets in a randomized block design for 35 d. Diets containing 16, 12, and 11% CP were fed in each experiment. All 12 and 11% CP diets were supplemented with lysine, tryptophan, threonine, and methionine to provide the same total concentrations as those in the 16% CP diet. In Exp. 1, the 11% CP diet was supplemented with isoleucine, valine, or isoleucine + valine to concentrations equal to those in the 16% CP diet. In Exp. 2, the 11% CP diet was supplemented with histidine, histidine + valine, or histidine + isoleucine + valine. In Exp. 3, the 11% CP diet was supplemented with valine, histidine + valine, or isoleucine + valine. Gilts were allowed free access to feed and water. In all experiments, ADG and feed efficiency (G/F) were reduced (P < or = 0.07) as dietary protein was reduced. Supplementation of isoleucine alone further reduced (P < 0.05) ADG, ADFI, G/F, and fat-free lean gain. In contrast, supplementation of valine alone resulted in numerical increases in ADG and ADFI in two experiments, although the differences were not significant (P > 0.05). Supplementation with histidine and valine together resulted in growth performance equal to or greater than that of pigs fed the 12% CP diet, but less than that of pigs fed the 16% CP diet. Supplementation of isoleucine and valine together resulted in better growth performance (P < 0.05) than supplementation of either amino acid alone. In two experiments (Exp. 1 and 3), supplementation of the 11% CP diet with isoleucine and valine together resulted in ADG that were not significantly different (P > 0.05) from those of pigs fed the 16% CP diet. Supplementation of all three amino acids (Exp. 2) did not improve performance over supplementations with histidine and valine. Plasma urea concentrations were reduced (P < 0.05) as dietary protein was lowered from 16 to 12%. Additions of crystalline amino acids did not affect plasma urea levels. Plasma amino acid concentrations reflected the dietary additions of crystalline amino acids, but did not assist in the identification of the sequence of limiting amino acids. These data suggest that valine is the fifth-limiting amino acid and that either histidine or isoleucine is the sixth-limiting amino acid in an 11% CP diet.

Nitrogen metabolism and growth performance of gilts fed standard corn-soybean meal diets or low-crude protein, amino acid-supplemented diets.

Two experiments were conducted to determine the CP concentration below which N retention and growth performance are reduced when low-protein, amino acid-supplemented, corn-soybean meal diets are fed. In a N balance trial (Exp. 1), 12 gilts (initial weight 41 kg) were fitted with urinary catheters and fed six different diets during three 7-d periods in an incomplete block design. The diets were: 1) 18% CP; 2) 14% CP + AA, 3) 16% CP; 4) 12% CP + AA; 5) 14% CP; and 6) 10% CP + AA. Amino acids (lysine, threonine, tryptophan, and methionine) were supplemented such that the concentrations in the low-protein diets were equal to those in their standard (4% CP higher) counterparts. Nitrogen retention (g/d) decreased (P < 0.01) as CP decreased, in both standard (27.10, 24.53, and 20.99) and low-protein (21.51, 19.18, and 15.83) diets, but was lower (P < 0.01) in low-protein diets. There were no differences among treatments (P > 0.05) in biological value (68.2% standard vs 71.0% low-protein). In a growth performance trial (Exp. 2), 36 gilts (initial weight 19.5 kg) were penned individually and fed one of six diets for 35 d in a randomized complete block design. Dietary treatments were a 16% CP standard diet and low-protein diets formulated to contain 15, 14, 13, 12, and 11% CP supplemented with crystalline lysine, tryptophan, threonine, and methionine to equal the total concentrations in the standard diet. Protein concentration affected (P < or = 0.05) ADG, ADFI, feed efficiency, fat-free lean gain, longissimus muscle area, plasma urea, and plasma concentrations of most essential AA. For most of these traits, the major difference was poor performance of pigs fed the 11% CP diet. Thus, in Exp. 1, at AA concentrations from deficient to excess, low-protein, amino acid-supplemented diets failed to produce the same N retention as the equivalent corn-soybean meal diets. However in Exp. 2, the same performance was obtained with 16, 15, 14, 13, and 12% CP. Based on these data, we suggest that N balance is more sensitive than growth to amino acid adequacy andthat other AA (e.g., isoleucine and valine) may limit growth performance when the protein concentration is reduced by more than four percentage units.

Discovery and evaluation of piperidinyl carboxylic acid derivatives as potent alpha(4)beta(1) integrin antagonists.

Piperidinyl carboxylic acid-based derivatives were prepared as antagonists of the leukocyte cell adhesion process that is mediated through the interaction of the alpha(4)beta(1) integrin (VLA-4, very late antigen 4) and the vascular cell adhesion molecule 1 (VCAM-1). Compounds 2a-h inhibited the adhesion in a cell-based assay in the low and sub micromolar range, a pharmacokinetic study of 2d is reported.

Tomato Ve disease resistance genes encode cell surface-like receptors.

In tomato, Ve is implicated in race-specific resistance to infection by Verticillium species causing crop disease. Characterization of the Ve locus involved positional cloning and isolation of two closely linked inverted genes. Expression of individual Ve genes in susceptible potato plants conferred resistance to an aggressive race 1 isolate of Verticillium albo-atrum. The deduced primary structure of Ve1 and Ve2 included a hydrophobic N-terminal signal peptide, leucine-rich repeats containing 28 or 35 potential glycosylation sites, a hydrophobic membrane-spanning domain, and a C-terminal domain with the mammalian E/DXXXLphi or YXXphi endocytosis signals (phi is an amino acid with a hydrophobic side chain). A leucine zipper-like sequence occurs in the hydrophobic N-terminal signal peptide of Ve1 and a Pro-Glu-Ser-Thr (PEST)-like sequence resides in the C-terminal domain of Ve2. These structures suggest that the Ve genes encode a class of cell-surface glycoproteins with receptor-mediated endocytosis-like signals and leucine zipper or PEST sequences.

Molecular characterization of a blue-copper laccase, TILA, of Aspergillus nidulans.

Laccases are blue-copper enzymes, which oxidize phenolic substrates and thereby reduce molecular oxygen. They are widespread within fungi and are involved in lignin degradation or secondary metabolism such as pigment biosynthesis. Many fungi contain several laccases, not all of whose functions are known. In Aspergillus nidulans one, yA, is expressed during asexual development and converts a yellow precursor to the green pigment. We identified a second laccase gene, which encodes a 66.3-kDa protein 37.6% identical to laccase I of A. nidulans. The protein harbors an N-terminal secretion signal, and three characteristic copper-binding centers. The enzyme localizes at the growing hyphal tip. The gene was therefore named tilA (=tip laccase). Deletion or overexpression of the gene had no discernible phenotype under laboratory conditions.

Dietary manipulation to reduce aerial ammonia concentrations in nursery pig facilities.

Two 4-wk trials (preliminary study) and three 5-wk trials (major study) were conducted to determine the effects of adding Yucca schidigera extract or anhydrous calcium chloride to nursery diets on the growth performance of nursery pigs and aerial ammonia concentration. The pigs were weaned between 13 and 15 d of age and had an initial BW of 3 to 6 kg. In each trial, pigs were allotted to three identical pig nursery rooms that were environmentally regulated. There were three diets (one diet per room): 1) control, containing 23% CP; 2) control plus 125 ppm of Yucca schidigera extract; and 3) control plus 1.95% anhydrous calcium chloride. Growth performance was recorded weekly. Aerial ammonia concentration was measured daily using aspiration detector tubes and during the last week of each trial using diffusion tubes. Manure samples were collected twice a week during the experimental period to determine ammonia and N concentrations and pH. Plasma urea concentration was determined in blood samples collected from the pigs at the end of each trial. Data were analyzed using split-plot and Latin square designs for the preliminary and major studies, respectively. Feed intake was similar among pigs fed all three diets. There were no differences in ADG and ADG/ADFI (G/F) between pigs fed the control diet and pigs fed the yucca extract diet (P > or = 0.41). In all trials, pigs fed the calcium chloride diet had lower ADG and G/F than pigs fed the other two diets (P < 0.05). In the preliminary study, aerial ammonia tended to be greater in the rooms in which pigs were fed the control diet than in the rooms with pigs fed the yucca extract diet (P = 0.08) and the calcium chloride diet (P = 0.11). In the major study, aerial ammonia increased weekly (diet x week; P < 0.001) in all rooms. In the 4th wk, ammonia concentrations were greater (P < 0.001) in the rooms in which pigs were fed the control diet than in the rooms in which the other two diets were fed. Dietary treatment had no effect on plasma urea concentration (P > or = 0.10), manure ammonia and N concentrations (P > or = 0.50), and manure pH (P > or = 0.78). Although aerial ammonia concentrations were relatively low, the addition of Yucca schidigera extract or calcium chloride to the diet of nursery pigs reduced ammonia concentrations in the nursery rooms.

Molecular farming of pharmaceutical proteins.

Molecular farming is the production of pharmaceutically important and commercially valuable proteins in plants. Its purpose is to provide a safe and inexpensive means for the mass production of recombinant pharmaceutical proteins. Complex mammalian proteins can be produced in transformed plants or transformed plant suspension cells. Plants are suitable for the production of pharmaceutical proteins on a field scale because the expressed proteins are functional and almost indistinguishable from their mammalian counterparts. The breadth of therapeutic proteins produced by plants range from interleukins to recombinant antibodies. Molecular farming in plants has the potential to provide virtually unlimited quantities of recombinant proteins for use as diagnostic and therapeutic tools in health care and the life sciences. Plants produce a large amount of biomass and protein production can be increased using plant suspension cell culture in fermenters, or by the propagation of stably transformed plant lines in the field. Transgenic plants can also produce organs rich in a recombinant protein for its long-term storage. This demonstrates the promise of using transgenic plants as bioreactors for the molecular farming of recombinant therapeutics, including vaccines, diagnostics, such as recombinant antibodies, plasma proteins, cytokines and growth factors.

Alterations in CER6, a gene identical to CUT1, differentially affect long-chain lipid content on the surface of pollen and stems.

Very long chain lipids contribute to the hydrophobic cuticle on the surface of all land plants and are an essential component of the extracellular pollen coat in the Brassicaceae. Mutations in Arabidopsis CER genes eliminate very long chain lipids from the cuticle surface and, in some cases, from the pollen coat. In Arabidopsis, the loss of pollen coat lipids can disrupt interactions with the stigma, inhibiting pollen hydration and causing sterility. We have positionally cloned CER6 and demonstrate that a wild-type copy complements the cer6-2 defect. In addition, we have identified a fertile, intragenic suppressor, cer6-2R, that partially restores pollen coat lipids but does not rescue the stem wax defect, suggesting an intriguing difference in the requirements for CER6 activity on stems and the pollen coat. Importantly, analysis of this suppressor demonstrates that low amounts of very long chain lipids are sufficient for pollen hydration and germination. The predicted CER6 amino acid sequence resembles that of fatty acid-condensing enzymes, consistent with its role in the production of epicuticular and pollen coat lipids >28 carbons long. DNA sequence analysis revealed the nature of the cer6-1, cer6-2, and cer6-2R mutations, and segregation analysis showed that CER6 is identical to CUT1, a cDNA previously mapped to a different chromosome arm. Instead, we have determined that a new gene, CER60, with a high degree of nucleotide and amino acid similarity to CER6, resides at the original CUT1 locus.