Biomedical subjects

D Krebs

Publications and source records attributed to D Krebs.

At least 19 recordsLinked to original sources

PubMed2001

Association of soluble HLA-G plasma levels with HLA-G alleles.

Soluble HLA-G (sHLA-G) molecules are found in the peripheral blood of healthy females and males, in cord blood and in amniotic fluids and discussed to be a mediator in maternal-fetal tolerance. In this study we investigated whether there are allele-specific differences in expression of sHLA-G molecules. For this, the sHLA-G plasma concentrations of 94 healthy unrelated individuals were measured by ELISA and correlated to their HLA-G genotypes, as determined by sequence analysis of exon 2 and 3 of the HLA-G gene. Mean sHLA-G levels in individuals with the most common HLA-G alleles G*01011 (27.0+/-2.1 SEM ng/ml, n=66), G*01012 (28.4+/-3.2 SEM ng/ml, n=34) were very similar. In contrast, individuals carrying the HLA-G*01013 (8.1+/-1.7 SEM ng/ml, n=17) or the "null" allele HLA-G*0105N (8.2+/-3.2 SEM ng/ml, n=7) presented significantly (P(c)=0.001 and P(c)<0.01, resp.) reduced sHLA-G levels. Furthermore, individuals with the HLA-G*01041 allele had significantly (P(c)=0.004) increased sHLA-G levels (42.5+/-4.6 SEM ng/ml, n=14). These results demonstrate that the generation of sHLA-G molecules is associated to certain HLA-G alleles and imply that sHLA-G levels are under genetic control. As low and high sHLA-G plasma levels did not segregate with HLA haplotypes including the HLA-G*01013 or *01041 allele, additional mechanisms may be involved in the regulation of the individual sHLA-G levels. Nevertheless, the existence of "low" and "high secretor" HLA-G alleles further suggests different levels of functionality in immune regulation.

Alleles↗

PubMed2000

Influence of salmeterol and benzalkonium chloride on G-protein-mediated exocytotic responses of rat peritoneal mast cells.

The long-acting beta(2)-adrenoceptor agonist salmeterol and the invert soap benzalkonium chloride share physicochemically important structures, namely a polar head group and a long aliphatic chain. Low concentrations of benzalkonium chloride have been shown to inhibit exocytotic responses in rat peritoneal mast cells by selectively interacting with heterotrimeric G-proteins of the G(i)-type. The present study investigates whether salmeterol inhibits, independently of beta-adrenoceptors, exocytotic responses of rat peritoneal mast cells induced by the direct agonists at G-proteins mastoparan or guanosine 5'-O-(3-thiotriphosphate) (++GTP gamma S++). Exocytosis was studied by secretion assays ([3H]5-hydroxytryptamine ([3H]5-HT)-release) using intact, streptolysin O-permeabilised or metabolically inhibited (antimycin, deoxyglucose) rat peritoneal mast cells. Both amphiphilics, salmeterol, and benzalkonium chloride, dose-dependently exerted biphasic effects on mastoparan-induced [3H]5-HT release in intact mast cells. In contrast to benzalkonium chloride, the dose-response curves for secretostatic and celltoxic effects of salmeterol markedly overlapped. Similar to benzalkonium chloride, salmeterol in non-cytotoxic concentrations (1-25 microg/ml) dose-dependently inhibited exocytosis induced by mastoparan (intact cells) or ++GTP gamma S (permeabilised cells). These findings indicate a direct, adrenoceptor-independent affection of G proteins by salmeterol in mast cells.

Adrenergic beta-Agonists↗

PubMed2000

Predicting the outcome of infertility surgery.

We studied a group of 187 infertile patients who had electromicrosurgery after laparoscopy, and falloposcopy. Follow-up information about 154 of these patients (82.4%) was available. Success rates in terms of intrauteine pregnancy was inversely related to the severity of peritubal and intratubal adhesions. Electromicrosurgery should only be preformed when clinical benefit is reasonably likely.

Adult↗

PubMed2000

Soluble HLA levels in early pregnancy after in vitro fertilization.

Intact pregnancy can be interpreted as a state of maternal immunotolerance toward an haploidentical fetus. Soluble HLA (sHLA) molecules increase during episodes of allograft rejection and are discussed as candidates to modulate immune responses. We questioned whether after in vitro fertilization (IVF) the subsequent intact pregnancy, early abortion, or tubal pregnancy influence the courses sHLA serum levels. Therefore, serum samples of 65 IVF patients were assayed by ELISA for sHLA-I, sHLA-G, and sHLA-DR concentrations preovulatorily and after a positive HCG test weekly until the 9th gestational week (GW). In 20 patients experiencing an early abortion the preovulatory sHLA-G mean level of 25.9 +/- 3.9 SEM ng/ml and the share of 4.2 +/- 0.8 SEM % on total sHLA-I were significantly (p < 0.05) reduced compared to women with intact pregnancy. The same differences (p < 0.0001) were seen during the monitoring of sHLA-G and sHLA-I levels in intact pregnancy versus early abortion until 9th GW. Twin pregnancy revealed a drastically increase of sHLA-G levels from the 8th GW compared to singleton pregnancies. Further, individual sHLA-DR levels increased during intact pregnancy but decreased in the group of early abortion. With regard to sensitivity and specificity for pregnancy outcome sHLA quantitation reached similar weight as routine HCG determinations at GW 5. Especially women with preovulatory low sHLA-G levels appear to be on risk for early abortion after IVF.

Abortion, Spontaneous↗

PubMed2000

[Prognostic value of Fallopian tube endoscopy].

OBJECTIVE: Progress in sterility-therapy demands diagnostic methods, that ensure the right indication for certain therapies in order to avoid unsuccessful attempts. In this retrospective study we tried to find out, whether we could predict and influence the success rate of therapies with the use of falloposcopy prior to further treatment. MATERIAL AND METHODS: In 62 sterile patients the endotubal state was evaluated with a falloposcope (Imagyn, Irvine, USA), a system of linear everting catheter (LEC) and microendoscope, prior to further treatment. RESULTS: Twenty of the 62 patients had endotubal pathology of both tubes, 15 patients showed pathology of one tube and 25 patients were diagnosed to have a normal endotubal morphology. In 2 patients falloposcopy could not be performed because of blockage due to intramural myomas. Pregnancy rate of those patients with a normal endosalpinx was 52% altogether, following microsurgery 80% and following insemination even 100%. No extrauterine pregnancies were described in this group. Pregnancy rate of those patients with endotubal pathology of both tubes was 35%. In this group no pregnancy followed microsurgery and following insemination one extrauterine pregnancy could be noticed. Furthermore, IVF results were improved if morphologic evaluation showed normal mucosa. CONCLUSIONS: The falloposcopy is a helpful diagnostic procedure to evaluate endotubal pathology prior to microsurgery and insemination.

Adult↗

PubMed2000

Evidence for major histocompatibility complex-mediated effects on spermatogenesis in humans.

Studies in fertile and infertile populations suggest an influence of genes of the major histocompatibility complex (MHC) on reproduction, although it remains unresolved if MHC-mediated effects on fertility are based on direct immunological or non-immunological effects of human leukocyte antigen (HLA) genes or rather on defects in MHC-associated non-HLA genes which affect gamete quality and embryonic development. We analysed allele frequencies for HLA class II loci DQA1, DQB1 and DRB1 and HLA class II haplotype frequencies in couples with tubal and andrological infertility who were treated with assisted reproductive techniques. Males with severe andrological infertility had significantly different allele frequencies for all three HLA loci when compared to males with normozoospermia, whereas no difference was found in the females. Differences in allele frequencies were stronger when only males whose partners achieved pregnancies after assisted reproduction treatment were compared. In those subgroups, we could also observe significant differences in three locus HLA class II haplotype frequencies. In summary, patients with male factor infertility differ in their HLA class II allele constitution from males with normozoospermia, which suggests that genes identical to or located in close vicinity to HLA class II genes may influence spermatogenesis and male gamete function.

Alleles↗

PubMed2000

Extracorporeal perfusion of the human uterus as an experimental model in gynaecology and reproductive medicine.

Experimental perfusion of various organs has primarily been used in transplantation medicine to study the physiology, pathophysiology and metabolism of tissues and cells. The purpose of this study was to establish an experimental model for the extracorporeal perfusion of the human uterus with recirculation of a modified, oxygenated Krebs-Henselait solution, in comparison with a non-recirculating perfusion system. With consent of the patients we obtained 25 uteri after standard hysterectomy. We performed an isovolumetric exchange of the perfusion medium at different intervals from 1 to 6 h and examined pH, pO(2), pCO(2), lactate, lactate dehydrogenase and creatine kinase by taking arterial and venous samples every hour for 24 h. We found the perfusions to be adequate when maintaining flow rates at 15-35 ml/min and at pressures ranging from 70 to 130 mmHg. Isovolumetric exchange of the perfusate every 3-4 h was the maximum interval to keep pH, the arterio-venous gradients of pO(2) and pCO(2), and the other biochemical parameters in physiological ranges. Examination by light and electron microscopy showed well-preserved features of myometrial and endometrial tissue. However, a 6 h exchanging interval led to increasing hypoxic and cytolytic parameters during the whole perfusion period. X-ray studies using digital subtraction angiography and perfusion studies with methylene blue demonstrated the homogeneous distribution of the perfusion fluid throughout the entire organ.

Adult↗

PubMed2000

Breast MR imaging screening in 192 women proved or suspected to be carriers of a breast cancer susceptibility gene: preliminary results.

PURPOSE: To compare magnetic resonance (MR) imaging with conventional imaging in screening high-risk women. MATERIALS AND METHODS: This prospective trial included 192 asymptomatic and six symptomatic women who, on the basis of personal or family history or genetic analysis, were suspected or proved to carry a breast cancer susceptibility gene. RESULTS: Fifteen breast cancers were identified: nine in the 192 asymptomatic women (six in the first and three in the second screening round) and six in the symptomatic patients. Concerning the asymptomatic women, four of the nine breast cancers were detected and correctly classified with mammography and ultrasonography (US) combined; another two cancers were visible as well-circumscribed masses and were diagnosed as fibroadenomas. MR imaging allowed the correct classification and local staging of all nine cancers. In 105 asymptomatic women with validation of the 1st-year screening results, the sensitivities of mammography, US, and MR imaging were 33%, 33% (mammography and US combined, 44%), and 100%, respectively; the positive predictive values were 30%, 12%, and 64%, respectively. CONCLUSION: The accuracy of MR imaging is significantly higher than that of conventional imaging in screening high-risk women. Difficulties can be caused by an atypical manifestation of hereditary breast cancers at both conventional and MR imaging and by contrast material enhancement associated with hormonal stimulation.

Adult↗

PubMed1999

Self-association and domains of interactions of an amphipathic helix peptide inhibitor of HIV-1 integrase assessed by analytical ultracentrifugation and NMR experiments in trifluoroethanol/H(2)O mixtures.

EAA26 (VESMNEELKKIIAQVRAQAEHLKTAY) is a better inhibitor of human immunodeficiency virus, type 1, integrase than its parent Lys-159, reproducing the enzyme segment 147-175 with a nonpolar-polar/charged residue periodicity defined by four helical heptads (abcdefg) prone to collapse into a coiled-coil. Circular dichroism, nuclear magnetic resonance, sedimentation equilibrium, and chemical cross-linking were used to analyze EAA26 in various trifluoroethanol/H(2)O mixtures. In pure water the helix content is weak but increases regularly up to 50-60% trifluoroethanol. In contrast the multimerization follows a bell-shaped curve with monomers in pure water, tetramers at 10% trifluoroethanol, and dimers at 40% trifluoroethanol. All suggest that interhelical interactions between apolar side chains are required for the coiled-coil formation of EAA26 and subsist at medium trifluoroethanol concentration. The N(H) temperature coefficients measured by nuclear magnetic resonance show that at low trifluoroethanol concentration the amide groups buried in the hydrophobic interior of four alpha-helix bundles are weakly accessible to trifluoroethanol and are only weakly subject to its hydrogen bond strengthening effect. The increased accessibility of trifluoroethanol to buried amide groups at higher trifluoroethanol concentration entails the reduction of the hydrophobic interactions and the conversion of helix tetramers into helix dimers, the latter displaying a smaller hydrophobic interface. The better inhibitory activity of EAA26 compared with Lys-159 could arise from its better propensity to form a helix bundle structure with the biologically important helical part of the 147-175 segment in integrase.

Amino Acid Sequence↗

PubMed1999

Precision contact of the fingertip reduces postural sway of individuals with bilateral vestibular loss.

Contact of the hand with a stationary surface attenuates postural sway in normal individuals even when the level of force applied is mechanically inadequate to dampen body motion. We studied whether subjects without vestibular function would be able to substitute contact cues from the hand for their lost labyrinthine function and be able to balance as well as normal subjects in the dark without finger contact. We also studied the relative contribution of sight of the test chamber to the two groups. Subjects attempted to maintain a tandem Romberg stance for 25 s under three levels of fingertip contact: no contact; light-touch contact, up to 1 N (approximately 100 g) force; and unrestricted contact force. Both eyes open and eyes closed conditions were evaluated. Without contact, none of the vestibular loss subjects could stand for more than a few seconds in the dark without falling; all the normals could. The vestibular loss subjects were significantly more stable in the dark with light touch of the index finger than the normal subjects in the dark without touch. They also swayed less in the dark with light touch than when permitted sight of the test chamber without touch, and less with sight and touch than just sight. The normal subjects swayed less in the dark with touch than without, and less with sight and touch than sight alone. These findings show that during quiet stance light touch of the index finger with a stationary surface can be as effective or even more so than vestibular function for minimizing postural sway.

Aged↗

PubMed1999

Serum oxytocin concentration during embryo transfer procedure.

OBJECTIVE: To determine the effect of the embryo transfer (ET) procedure on serum concentration of oxytocin. STUDY DESIGN: Prospective clinical study of 10 women undergoing in vitro fertilization (IVF) treatment with ET in the Section of Reproductive Medicine and Endocrinology at the Department of Obstetrics and Gynecology, University of Bonn, Germany. Serial blood samples were collected in time intervals of 20 s during embryo transfer procedure and serum oxytocin concentration was measured. RESULTS: In the absence of tenaculum placement, none of the procedures associated with ET led to an increase in serum oxytocin concentration. When tenaculum placement was used, it was temporally (four out of five patients) associated with an elevation in oxytocin level, which remained elevated until of the end of ET procedure. CONCLUSION: Application of a cervical tenaculum during ET or possibly also during intra uterine insemination (IUI) procedure can stimulate the release of oxytocin in some patients.

Adult↗

PubMed1999

Exclusion of a major role for the PTEN tumour-suppressor gene in breast carcinomas.

PTEN is a novel tumour-suppressor gene located on chromosomal band 10q23.3. This region displays frequent loss of heterozygosity (LOH) in a variety of human neoplasms including breast carcinomas. The detection of PTEN mutations in Cowden disease and in breast carcinoma cell lines suggests that PTEN may be involved in mammary carcinogenesis. We here report a mutational analysis of tumour specimens from 103 primary breast carcinomas and constitutive DNA from 25 breast cancer families. The entire coding region of PTEN was screened by single-strand conformation polymorphism (SSCP) analysis and direct sequencing using intron-based primers. No germline mutations could be identified in the breast cancer families and only one sporadic carcinoma carried a PTEN mutation at one allele. In addition, all sporadic tumours were analysed for homozygous deletions by differential polymerase chain reaction (PCR) and for allelic loss using the microsatellite markers D10S215, D10S564 and D10S573. No homozygous deletions were detected and only 10 out of 94 informative tumours showed allelic loss in the PTEN region. These results suggest that PTEN does not play a major role in breast cancer formation.

BRCA2 Protein↗

PubMed1999

Conformational aspects of HIV-1 integrase inhibition by a peptide derived from the enzyme central domain and by antibodies raised against this peptide.

Monospecific antibodies were raised against a synthetic peptide K159 (SQGVVESMNKELKKIIGQVRDQAEHLKTA) reproducing the segment 147-175 of HIV-1 integrase (IN). Synthesis of substituted and truncated analogs of K159 led us to identify the functional epitope reacting with antibodies within the C-terminal portion 163-175 of K159. Conformational studies combining secondary structure predictions, CD and NMR spectroscopy together with ELISA assays, showed that the greater is the propensity of the epitope for helix formation the higher is the recognition by anti-K159. Both the antibodies and the antigenic peptide K159 exhibited inhibitory activities against IN. In contrast, neither P159, a Pro-containing analog of K159 that presents a kink around proline but with intact epitope conformation, nor the truncated analogs encompassing the epitope, were inhibitors of IN. While the activity of antibodies is restricted to recognition of the sole epitope portion, that of the antigenic K159 likely requires interactions of the peptide with the whole 147-175 segment in the protein [Sourgen F., Maroun, R.G., Frère, V., Bouziane, A., Auclair, C., Troalen, F. & Fermandjian, S. (1996) Eur. J. Biochem. 240, 765-773]. Actually, of all tested peptides only K159 was found to fulfill condition of minimal number of helical heptads to achieve the formation of a stable coiled-coil structure with the IN 147-175 segment. The binding of antibodies and of the antigenic peptide to this segment of IN hampers the binding of IN to its DNA substrates in filter-binding assays. This appears to be the main effect leading to inhibition of integration. Quantitative analysis of filter-binding assay curves indicates that two antibody molecules react with IN implying that the enzyme is dimeric within these experimental conditions. Together, present data provide an insight into the structure-function relationship for the 147-175 peptide domain of the enzyme. They also strongly suggest that the functional enzyme is dimeric. Results could help to assess models for binding of peptide fragments to IN and to develop stronger inhibitors. Moreover, K159 antibodies when expressed in vivo might exhibit useful inhibitory properties.

Amino Acid Sequence↗

PubMed1999

Pain palliation with rhenium-186 HEDP in breast cancer patients with disseminated bone metastases.

PURPOSE: For patients with metastatic prostate cancer, first results have shown that rhenium-186 (Re-186) hydroxyethylidene diphosphonate (HEDP) is efficient in pain palliation of disseminated bone metastases. The aim of this study was to determine whether significant pain reduction can also be achieved in breast cancer patients with Re-186 HEDP. METHODS: Thirty patients with breast cancer who had multifocal painful bone metastases received a total of 38 intravenous Re-186 HEDP injections. Pain relief was assessed through daily documentation of the visual analog scale and analgesic consumption. A significant response to treatment was determined if the visual analog scale or analgesic consumption decreased significantly for at least 2 weeks. Blood counts were controlled at baseline and at weeks 4 and 8. RESULTS: A response to pain therapy was observed in 60% (18 of 30) of the patients. A reversible thrombocytopenia and leukopenia of grade 2 (according to World Health Organization criteria) was found in 4 and 2 patients, respectively. CONCLUSION: Patients with disseminated osseous metastases resulting from breast cancer can benefit from therapy with Re-186 HEDP.

Adult↗

PubMed1999

Exercise--it's never too late: the strong-for-life program.

OBJECTIVES: This investigation determined whether an in-home resistance training program achieved health benefits in older adults with disabilities. METHODS: A randomized controlled trial compared the effects of assigning 215 older persons to either a home-based resistance exercise training group or a waiting list control group. Assessments were conducted at baseline and at 3 and 6 months following randomization. The program consisted of videotaped exercise routines performed with elastic bands of varying thickness. RESULTS: High rates of exercise adherence were achieved, with 89% of the recommended exercise sessions performed over 6 months. Relative to controls, subjects who participated in the program achieved statistically significant lower extremity strength improvements of 6% to 12%, a 20% improvement in tandem gait, and a 15% to 18% reduction in physical and overall disability at the 6-month follow-up. No adverse health effects were encountered. CONCLUSIONS: These findings provide important evidence that home-based resistance exercise programs designed for older persons with disabilities hold promise as an effective public health strategy.

Aged↗

PubMed1999

[Monoclonal anti-idiotype antibodies in immunotherapy of ovarian carcinoma (MAb ACA125) and breast carcinoma (MAb ACA14C5)].

Anti-idiotypic antibodies, which imitate a tumor-associated antigen by their variable region, offer an elegant method for the induction of a specific immune response, when used as a surrogate antigen for immunization. We generated anti-idiotypic antibodies imitating 2 different tumor-associated antigens. I. CA125 for ovarian carcinomas and II. 14C5, a tumor-associated cell substrate adhesion molecule on breast cancer cells, whereas the first approach could be introduced in a first clinical trial and the second was evaluated in an immunocompetent animal model. For the induction of an immune response against CA125, 18 patients with advanced ovarian cancer (n = 6) or heavily pretreated recurrences (n = 12) were immunized with the anti-idiotypic antibody MAb ACA125. Patients were treated with 2 mg anti-idiotype antibody every two weeks for 4 injections i.m. and then monthly. 12 of 18 patients demonstrated an anti-anti-idiotypic (Ab3) response, which was to a lower extent also directed against CA125 and 9 of 18 patients developed a CA125 specific cellular immune response by their peripheral blood lymphocytes. Based on this data a follow-up clinical trial in advanced ovarian cancer patients with minimal residual disease in an adjuvant approach after primary therapy was started to evaluate the effect of the immune response on the progression free survival. For immunotherapy of breast cancer, we generated a murine monoclonal anti-idiotypic antibody (MAb ACA14C5), which imitates a cell substrate adhesion molecule on breast cancer cells. The anti-idiotype was introduced in an immunocompetent animal to prove his capability on induction of an immune and tumor response. The results showed a highly significant difference in the tumor growth of the ACA14C5 treated group in contrast to the controls starting the immunization on day 6 after tumor cell application with 10 of 12 animals being cured from their tumor burden. Prophylactic immunization against the invasion antigen of breast cancer by anti-idiotypic antibodies showed protection against increasing tumor burden. However, in the situation of established tumors only minor responses could be detected. Vaccination with anti-idiotypic antibodies comprises an effective method for induction of a specific immune response against non-immunogenic tumor-associated antigens and should be therefore considered in immunological approaches to tumor therapy, where the primary structure and sequence of the antigen, e.g. CA125, is up to now not available.

Antibodies, Monoclonal↗

PubMed1999

Clinical application of maternal serum cytokine determination in premature rupture of membranes--interleukin-6, an early predictor of neonatal infection?

BACKGROUND: In cases of premature rupture of membranes (PROM), an early detection of fetal infection is necessary in order to weigh infectious complications against prematurity. As routine parameters (leukocytes, C-reactive protein (CRP), fever, and fetal tachycardia) lack satisfactory sensitivity and specificity, this study evaluates whether the determination of interleukin-6 (IL-6), interleukin-8 (IL-8) or soluble interleukin-2 receptor (IL-2R) in maternal serum could supplement or replace routine inflammation parameters. METHODS: In this prospective study results of clinical and laboratory parameters were investigated with respect to neonatal infection in 71 patients with PROM. IL-6, IL-8 and IL-2R were determined by enzyme immunoassays. RESULTS: Best specificity and sensitivity could be demonstrated for CRP and IL-6. Both elevation of CRP and IL-6 correlated significantly (p<0.01 and p<0.001, respectively) with the onset of neonatal infection. At a cutoff of 11 pg/ml, IL-6 reaches a sensitivity of 81% and a specificity of 76%; CRP a specificity of 76% (cutoff 1.2 mg/dl) and a sensitivity of 56%. In 4/16 (25%) cases developing neonatal infection, IL-6 increased earlier than CRP. IL-8 and IL-2R results showed a less significant correlation with fetal outcome. CONCLUSIONS: Determination of IL-6 in maternal serum can significantly contribute to an earlier detection of fetal infection in patients with PROM.

Area Under Curve↗

PubMed1998

Expression of the ATM gene is significantly reduced in sporadic breast carcinomas.

The gene mutated in ataxia telangiectasia (A-T) patients (ATM) is located on chromosome 11q22-23, a region frequently altered in mammary tumors. Patients homozygous for ATM mutations are prone to develop a variety of different neoplasms. Female heterozygotes have been reported to carry a 5- to 8-fold increased risk of breast cancer. However, germline mutations in the ATM gene are rare in women with sporadic breast carcinomas. Most of the alterations described in A-T patients result in a functionally inactive ATM protein. Moreover, it has been suggested that mutations of the ATM gene in A-T patients influence the amount of ATM mRNA and that this may affect the severity of the disease. In the present study, we have analyzed ATM transcripts in a series of 39 breast carcinomas, 14 benign breast lesions and 12 normal breast tissue samples. ATM mRNA levels were determined by semiquantitative competitive RT-PCR. Competitor RNA molecules for the ATM gene and the housekeeping gene beta-2-microglobulin (B2M) were generated by PCR mutagenesis. Low concentrations of ATM transcripts were detected in breast carcinomas, intermediate levels in benign lesions and highest levels in normal breast tissue specimens (F-test, p = 0.0013). Our results indicate that reduced expression of the ATM gene may contribute to the development and/or malignant progression of breast carcinomas.

Ataxia Telangiectasia Mutated Proteins↗