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The eukaryotic translation initiation factor 4E controls lettuce susceptibility to the Potyvirus Lettuce mosaic virus.

The eIF4E and eIF(iso)4E cDNAs from several genotypes of lettuce (Lactuca sativa) that are susceptible, tolerant, or resistant to infection by Lettuce mosaic virus (LMV; genus Potyvirus) were cloned and sequenced. Although Ls-eIF(iso)4E was monomorphic in sequence, three types of Ls-eIF4E differed by point sequence variations, and a short in-frame deletion in one of them. The amino acid variations specific to Ls-eIF4E(1) and Ls-eIF4E(2) were predicted to be located near the cap recognition pocket in a homology-based tridimensional protein model. In 19 lettuce genotypes, including two near-isogenic pairs, there was a strict correlation between these three allelic types and the presence or absence of the recessive LMV resistance genes mo1(1) and mo1(2). Ls-eIF4E(1) and mo1(1) cosegregated in the progeny of two separate crosses between susceptible genotypes and an mo1(1) genotype. Finally, transient ectopic expression of Ls-eIF4E restored systemic accumulation of a green fluorescent protein-tagged LMV in LMV-resistant mo1(2) plants and a recombinant LMV expressing Ls-eIF4E degrees from its genome, but not Ls-eIF4E(1) or Ls-eIF(iso)4E, accumulated and produced symptoms in mo1(1) or mo1(2) genotypes. Therefore, sequence correlation, tight genetic linkage, and functional complementation strongly suggest that eIF4E plays a role in the LMV cycle in lettuce and that mo1(1) and mo1(2) are alleles coding for forms of eIF4E unable or less effective to fulfill this role. More generally, the isoforms of eIF4E appear to be host factors involved in the cycle of potyviruses in plants, probably through a general mechanism yet to be clarified.

Amino Acid Sequence↗

Induction of transplantation immunity to rat colon carcinoma isografts by implantation of intact fetal colon tissue.

The objective of this study was to demonstrate whether intact fetal colon tissue can induce immunity to large-bowel cancer isografts. Adult rats were immunized against fetal antigens by implantation of mitomycin-C-treated intact fetal colon tissue beneath the renal capsule. Control animals received implants of pieces of intact adult colon tissue. Kidneys containing the implants were removed after 3 or 5 weeks and the rats were challenged with isografts of a 1,2-dimethylhydrazin (DMH)-induced colon carcinoma. Significant inhibition of tumor growth was observed in immunized groups compared to controls. The present results indicate that some embro-fetal antigens expressed in intact colon tissue of 13 to 15-day-old fetuses are also expressed in colon carcinoma cells and can act as transplantation antigens.

Animals↗

Regional scleral changes in form-deprivation myopia in chicks.

Similar neurochemical events appear to be involved in the development of myopia in chicks and mammals. The rapid post-hatching development of the chick is ideal for studying experimental myopia. In this investigation, one eye of 2-day-old chicks was deprived of form vision for 2 weeks and then compared to the fellow, non-deprived eye by immunohistochemistry and light and electron microscopy. All deprived eyes showed a high refractive error and ocular enlargement. In deprived eyes, the posterior cartilaginous sclera was thicker and the fibrous sclera of the same section was thinner than the control. Scleral morphological changes were restricted to a central button 6-7 mm in diameter (the posterior pole) within the posterior hemisphere, further divided into posterotemporal and posteronasal parts. The most enlarged, posterior cartilaginous structure of deprived sclera could be divided into an inner and an outer zone. The inner zone had many unevenly-arranged chondrocytes, each having a well-developed granular endoplasmic reticulum and Golgi complex and a very irregular cell surface. Numerous S-phase cells and isogenous groups were detected in the outer zone. Hypertrophic chondrocytes were often observed in the innermost region of the outer zone and the outermost region of the inner zone. The boundary between the outer fibrous sclera and the cartilaginous sclera was irregular and obscured in myopic eyes. Spindle-shaped chondrocytes were seen to be in contact with each other. Thick collagen fibrils, usually seen only in the outer fibrous sclera, were present among the chondrocytes. Results of this morphological study suggest an increased proliferation of chondrocytes and active synthesis of extracellular matrix in visually deprived eyes. The elongation of the ocular axis that accompanies myopia is caused primarily by an active remodeling and differentiation in a restricted section of the posterior scleral cartilage. These facts indicate the posterior scleral cartilage may be more immature than cartilage in anterior and lateral segments.

Animals↗

Recovery of the immune system in diabetic mice after transplantation of isolated islets of Langerhans.

Adult CBA/HZgb mice islets harvested by collagenase digestion were injected intraperitoneally in 52 singeneic diabetic recipients (CBA/Hgb leads to CBA/HZgb, 450-600 islets per mouse). Normal serum glucose levels, 24-h urine volume, insulin levels and body weight were completely restored to normal in all recipients during the next 2-5 months. Immunological function was assessed in control, diabetic and diabetic-transplanted mice by following their responses to sheep erythrocytes (expressed as the number plaque-forming cells in the spleen). In transplanted mice, the plaque-forming cell responses were as follows: 1 month after transplantation--43% of the plaque-forming cell counts in control (normal, non-diabetic) mice; 2 months after transplantation--56% of the control value; and 94% of the control value after 5 months. Ten months after the transplantation, the plaque-forming cell counts were slightly above the control value (148%). It appears, therefore that transplantated islet tissue positively affects the immunological as well as the diabetic state of the recipients.

Animals↗

The long-term metabolic function of intraportal and renal subcapsular islet isografts and the effect on glomerular basement membrane thickness in rats.

Previous studies of intraportal islet autotransplantation in large animals have reported graft failure after months or years. In the rat it has been reported that intraportal islet isografts eventually failed whilst islets transplanted to the renal subcapsule functioned up to a year. We made Dark Agouti (DA) rats severely diabetic with streptozotocin, then 1000 or 3000 DA islets were transplanted beneath the renal capsule or into the liver. One set of transplanted rats and untreated diabetic and normal non-diabetic littermates were monitored lifelong by measurement of plasma glucose, others were killed at 6, 12 and 18 months for measurement of haemoglobin A1c, intravenous glucose tolerance test, pancreas insulin content and histology of the kidney. Renal glomerular basement membrane thickness was measured by the orthogonal intercept method. The results showed that intraportal isografts reversed hyperglycaemia significantly faster than renal subcapsular isografts. In the renal subcapsular site, consistent reversal of diabetes was achieved with 3000 islets but not with 1000 islets. Furthermore, intraportal islet grafts with 3000 islets led to lower, normal random glucose level than renal subcapsular grafts for the first 13 months. Normoglycaemia was maintained lifelong in all rats that achieved early normoglycaemia after transplantation of 3000 islets, irrespective of the site of islet transplantation. The fasting glucose, haemoglobin A1c levels, K value and glomerular basement membrane thickness of the recipients of 3000 islets to either the intraportal and subcapsular site were not significantly different from each other and the normal controls up to 18 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of sequential multiple grafting of syngeneic newborn thymus on the immune functions and life expectancy of aging mice.

Enhancement of the immune functions and extension of the mean life expectancy were successfully performed in aging mice by sequential multiple grafting of syngeneic newborn thymus. In the first experiment, 2-month-old female C57BL/6 mice were grafted with either syngeneic newborn thymus or newborn spleen every 2 months, 5 or 6 times. A significant enhancement of T cell dependent immune functions were observed in the group sequentially grafted with newborn thymus, in comparison to that grafted with multiple sequential newborn spleen or with a single newborn thymus and that without a graft. In the second experiment, the same sequential grafting protocol was performed in middle aged mice at monthly interval for 4-5 consecutive months and the immune functions and survival rate were compared between the experimental and control groups. The immune functions were only partially rejuvenated, but an extension of the mean remaining life expectancy was observed in the experimental group (312 +/- 38 days) as compared with control (214 +/- 42 days), although maximal life-span was the same in both groups (1100 days).

Aging↗

Host-parasite interactions in Staphylococcus aureus keratitis.

Ulcerative keratitis is among the leading ocular bacterial infections, and Streptococcus aureus accounts for approximately 25% of cases in some surveys. Although S. aureus expresses numerous virulence factors, many of which are under the control of staphylococcal global regulatory genes, their pathophysiologic roles in keratitis are largely unknown. Similarly, the nature of the host response during S. aureus keratitis is unclear. Following a review of previously published research on the pathophysiology of S. aureus ocular infection, we present the results of a study designed to assess the host-parasite relationship between S. aureus and human corneal epithelial cells (HCECs) in vitro. In this model system, a wild-type S. aureus strain and its isogenic mutants harboring mutations in agr and sar global regulatory genes or fibronectin-binding proteins A and B (fnbAB) were tested for their ability to bind and invade confluent HCEC monolayers. The contribution of host cell factors was assessed by preincubating HCECs with various inhibitory agents. These studies demonstrated that S. aureus not only adhered to the surface of HCECs but was also internalized, as has been previously observed in other nonocular cell lines. Adherence and invasion of HCECs was saturable at 1 h of incubation in the presence of approximately 10(7) CFU per HCEC monolayer (multiplicity of infection approximately 10). A mutant defective in both agr and sar global regulators was not significantly different in invasive capacity compared to its isogenic wild-type parent strain. In contrast, mutations in fibronectin-binding proteins A and B (fnbAB) reduced the invasiveness of S. aureus by 99% compared to the wild-type strain. Pretreatment of HCECs with colchicine had little effect on S. aureus invasion. In sharp contrast, cytochalasin D and genistein were each capable of inhibiting invasion by >99%. In summary, the results of this study point to fibronectin-binding protein as a key S. aureus surface adhesin facilitating invasion of HCECs in vitro. Furthermore, these results suggest an active mechanism for S. aureus internalization by HCECs, likely involving actin polymerization and tyrosine kinase activity. Additional studies are warranted to determine the applicability of these findings in vivo, and to facilitate the rational design of therapeutic agents aimed at blocking the establishment and progression of S. aureus keratitis.

Adhesins, Bacterial↗

Characterization and growth in human macrophages of Mycobacterium avium complex strains isolated from the blood of patients with acquired immunodeficiency syndrome.

Strains of the Mycobacterium avium complex (MAC) yield opaque and transparent colonial variants when cultivated in vitro. The transparent variants are more virulent than the opaque for animals, but little is known about the respective roles of these colonial variants in humans. To assess which variant infects humans, various blood fractions from eight patients with MAC bacteremia were plated directly onto 7H10 agar. In cell fractionation studies, all the M. avium complex CFU were associated with leukocytes and none were found free in plasma. All colonies on the primary culture plate exhibited the transparent phenotype. However, during subculture in 7H9 broth or on Lowenstein-Jensen agar, opaque variants appeared in seven of eight strains. Isogenic pairs of transparent and opaque variants were prepared and used to infect in vitro human monocyte-derived macrophages from healthy seronegative individuals. Transparent variants invariably grew inside macrophages, but only one of seven opaque variants did so. These observations indicate that the bacteremia of M. avium complex in acquired immunodeficiency syndrome patients consists exclusively of the transparent variants, perhaps because these variants are able to multiply inside macrophages. In contrast, opaque variants appear after in vitro subculture and are controlled by human macrophages, consistent with their reduced virulence in animals.

Acquired Immunodeficiency Syndrome↗

Gene expression profiling in AGS cells stimulated with Helicobacter pylori isogenic strains (cagA positive or cagA negative).

To study host response to CagA, human gastric cancer cell line AGS was infected with a Helicobacter pylori type I wild-type or isogenic cagA-negative mutant. Differentially expressed genes were identified using cDNA array technology. By Northern blotting, downregulation of focal adhesion kinase and upregulation of LIM kinase mRNA in the presence of CagA were clearly verified. Furthermore, upregulation of LIM kinase, macrophage inflammatory protein-2, c-myc, and bone morphogenetic protein-1 and downregulation of transcription factor Y-box binding protein-1 and focal adhesion kinase mRNA in response to H. pylori type I infection compared to the uninfected control could be shown by Northern blotting. Hence, these findings identified new targets for further functional studies on H. pylori-associated pathogenesis.

Antigens, Bacterial↗

The role of autosensibilization of lymphoid cells in genesis of antitumor resistance.

C3HA mice were inoculated with a mixture of syngeneic transplantable hepatoma 22a cells and syngeneic splenocytes of mice immunized with normal syngeneic tissues or subjected to partial hepatectomy. Control mice were inoculated with a mixture of tumor cells and splenocytes of intact or sham operated mice. A considerable inhibition of tumor growth was observed in the experimental series. Immunization of mice with normal syngeneic tissues as well as partial hepatectomy results in sensibilization of splenocytes not only to normal tissue antigens, but to the antigens of hepatoma 22a cells too. This was shown by the reaction of indirect inhibition of glass adherence of peritoneal cells. The data obtained are considered to be one more possible prove of a special form immune surveillance which controls the state of cytodifferentiation being active in the organism. The involvement of this form of immune surveillance in the genesis of antitumor resistance is discussed.

Animals↗

The segregation of mitochondrial genes in yeast. II. Analysis of zygote pedigrees of drug-resistant X drug-sensitive crosses.

Cytoplasmically inherited chloramphenicol- and erythromycin-resistant mutants were obtained in three unrelated and two isogenic haploid strains of yeast. The bias favoring the transmission of these resistance alleles in crosses to the isogenic strains was compared on two levels: on the population level by means of observing random diploid progeny from mass matings, and on the zygote level by zygotic pedigree analyses. The genetic basis of this bias was determined by tetrad analysis. Our results suggest that 1. an intracellular selection mechanism operates within zygotes to determine the degree of bias; 2. the selection mechanism operates differently with respect to the two loci, C and E, under consideration; and 3. the selection mechanism is controlled by a set of nuclear genes. Other models which have been suggested to explain bias are critically examined in light of our results.

Chloramphenicol↗

DeltaMGMT-transduced bone marrow infusion increases tolerance to O6-benzylguanine and 1,3-bis(2-chloroethyl)-1-nitrosourea and allows intensive therapy of 1,3-bis(2-chloroethyl)-1-nitrosourea-resistant human colon cancer xenografts.

O6-Benzylguanine (BG) is a potent inhibitor of the DNA repair protein 06-alkylguanine DNA alkyltransferase (AGT), and sensitizes tumors to BCNU in vitro and in xenografts. The combination of BG and BCNU is now undergoing phase I clinical testing. The maximally tolerated dose of BCNU given after BG is expected to be lower then the doses tolerated as a single agent owing to BG sensitization of hematopoietic progenitors. We have previously shown that retroviral expression of G156A mutant MGMT (deltaMGMT) in mouse and human marrow cells results in significant BG and BCNU resistance. In this study we evaluated the effect of deltaMGMT-transduced marrow infusion on the therapeutic index of multiple BG and BCNU treatments in tumor-bearing nude (nu/nu athymic) mice. Prior to subcutaneous implantation of BCNU-resistant SW480 human colon cancer cells, cohorts of mice were given intraperitoneal injections of nonablative doses of BG (30 mg/kg) and BCNU (10 mg/kg, one-half of the LD10) and then infused with 1-2 x 10(6) isogeneic deltaMGMT (n = 29 mice) or lacZ-transduced (n = 20 mice) marrow cells. The xenograft-bearing mice were treated with multiple cycles of BG (30 mg/kg) and BCNU (10-25 mg/kg). After three cycles, deltaMGMT mouse bone marrow was repopulated with CFU containing the provirus, and demonstrated a 2.7-fold increase in AGT activity and a 5.5-fold increase in BCNU IC90 compared with LacZ mice. After five cycles, the BCNU IC90 of CFU cells increased nine-fold over control cells, indicating selective enrichment of CFU precursor cells expressing high levels of deltaMGMT. Starting with the third cycle of therapy, tolerance to BG and BCNU was significantly improved in deltaMGMT mice compared with LacZ mice, as evidenced by preserved peripheral blood counts, bone marrow cellularity, and CFU content 1 and 2 weeks posttreatment and a significantly higher survival rate. Xenograft growth was significantly delayed in mice tolerating multiple cycles and higher dose intensity of BG and BCNU as compared with mice receiving less intensive therapy. We conclude that deltaMGMT-transduced marrow cells can improve the therapeutic index of BG and BCNU by selectively repopulating the marrow and providing significant marrow tolerance to this combination, allowing intensive therapy of a BCNU-resistant tumor.

Animals↗

Reimplantation injury after lung transplantation in a rat model.

Distant procurement of organs for transplantation requires satisfactory preservation to reduce injury during ischemia and the initial phase of reperfusion. We have studied the mechanism of reimplantation injury after unilateral lung transplantation in isogeneic Fisher rats. The heart and lungs were removed en bloc from donor rats and preserved at 4 degrees C. After 5 hours the left lung was transplanted into a recipient. Radiographic and histologic evidence of pulmonary edema in the transplanted lung at 24 hours confirmed the presence of lung vascular injury. In five rats we performed bronchoalveolar lavage (BAL) of both nontransplanted and transplanted lungs at 24 hours posttransplant, immediately after the animal was killed. Results were compared with five normal control lungs. The results showed not only significantly greater number of cells from transplanted lungs compared with nontransplanted and control lungs but cell profiles showed much greater percentages of neutrophils (mean +/- SD) from transplanted (76.8% +/- 13%) compared with nontransplanted (2.8% +/- 3.1%) or control (0.8% +/- 0.8%) lungs. In seven other rats we measured BAL neutrophil activity with stimulated luminol chemoluminescence from transplanted left and nontransplanted right lungs 24 hours after unilateral left lung transplantation. Results (expressed as millivolt X 10(3) neutrophil +/- SD) showed significantly greater activity from transplanted (2.8 +/- 1.7) compared with nontransplanted (0.72 +/- 0.6) lungs. Reimplantation injury of the lung is characterized by pulmonary sequestration of neutrophils, and these cells may play a primary role in mediating vascular damage.

Animals↗

Short-term impact of carbon dioxide, helium, and gasless laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient.

BACKGROUND: Laparoscopic donor nephrectomy has the potential to increase the number of living kidney donations by reducing donor morbidity. However, studies have shown that raised intraabdominal pressure can result in transient renal dysfunction. Therefore, laparoscopically procured kidneys might be at higher risk for suffering a period of ischemia during pneumoperitoneum. The objective of this study was to investigate the short-term impact of pneumoperitoneum used for laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient. METHODS: EXPERIMENT 1: KIDNEY DONOR: Initially, 36 brown Norway (BN) rats were randomized for three procedures: 2 h of carbon dioxide (CO2) insufflation (8 mmHg), 2 h of helium insufflation (8 mmHg), and 2 h of gasless technique (0 mmHg). After this, a unilateral nephrectomy was performed in all the animals. EXPERIMENT 2: RECIPIENT: Subsequently, 36 donor BN rats were subjected to a similar insufflation protocol, but after nephrectomy, a syngeneic kidney transplantation (BN-BN) was performed. Urine and blood samples were collected on postoperative days 1, 3, 7, and 14 for determination of renal function. Subsequently, donor and recipient kidneys were removed for histomorphologic and immunohistochemical analysis. RESULTS: In both donors and recipients, no significant changes in serum creatinine, proteinuria, or glomular filtration were detected between the CO2, the helium, and the gasless control groups. In both experiments, histologic analysis of Kidney specimens did not show any deleterious effects from abdominal gas insufflation. Although kidney grafts exposed to CO2 showed significantly higher numbers of CD45+ leukocytes 3 days after transplantation, immunohistochemical analysis did not show significant differences in number of infiltrating cells (CD4, CD8, ED1, OX6, OX62) between the two insufflation groups and the gasless control subjects. CONCLUSIONS: Abdominal gas insufflation does not have an adverse effect on the renal function of the kidney donor 1 week after laparoscopic donor nephrectomy. No differences in renal function or histomorphology were detected between syngeneic kidney grafts exposed to pneumoperitoneum and gasless control subjects.

Animals↗

A new model for the study of mild diabetes during pregnancy. Syngeneic islet-transplanted STZ-induced diabetic rats.

Diabetes during pregnancy carries short- and long-term consequences for the offspring. Improved obstetrical and diabetic care has resulted in decreased morbidity and mortality in the neonate of the diabetic mother. Mild hyperglycemia is still found in both IDDM pregnant women and women with GDM. The long-term consequences of exposure to mild hyperglycemia in utero remain to be determined. In an effort to develop an appropriate animal model of mild diabetes during pregnancy, we mated female STZ-induced diabetic rats previously transplanted with specific numbers of islets of Langerhans (2500, 1000, 700, or 500 islets). Diabetic and nondiabetic sham-transplanted control groups also were studied. During pregnancy, the plasma glucose levels in the diabetic rats and the group receiving 500 islets (26.5 +/- 1.1 and 10.0 +/- 0.8 mM, respectively) were significantly greater than in control animals (5.4 +/- 0.5 mM, P < 0.0001). The mean glucose levels in rats receiving 700 or 1000 transplanted islets (6.8 +/- 0.2 and 6.5 +/- 0.2 mM) also were significantly greater than in control animals (P < 0.001). No difference was evident between control rats and the group receiving 2500 islets (5.8 +/- 0.2 mM). No gross congenital abnormalities were apparent in the offspring. The pup plasma glucose was significantly greater in the offspring of dams receiving either none (diabetic) or 500 islets (10.6 +/- 0.7 and 11.1 +/- 1.1 mM, respectively) compared with the offspring of nondiabetic control dams (4.4 +/- 0.3 mM, P < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Factors that control the differentiation of stem cells. I. The change in direction of hematopoietic stem cell differentiation under the effect of differentiating T-lymphocytes].

A mixed transplantation of bone marrow cells, and lymph nodes or thymic cells of mice CBA strain into lethally irradiated hybrid recipients (CBAXC57B1)F1 is accompanied with changes in the differentiation pattern from a mainly erythroid to a mainly granuloid way. Thymectomy of either donor of bone marrow cells or recipients, or both, destroys the stem cell differentiation in the direction of granulopoieseis. Intact syngeneic lymphocytes normalize differentiation of the stem cells, but in the presence of tissue antigens these provide for the stem cell differentiation mainly in the direction of granulopoiesis. The differentiation of stem haemopoietic cells is accomplished under the thymic and lymphocyte control. T-differentiating lymphocytes (Td) are the lymphocytes controlling the stem cell differentiation.

Animals↗

Bundle-forming pili and EspA are involved in biofilm formation by enteropathogenic Escherichia coli.

Microcolony formation is one of the initial steps in biofilm development, and in enteropathogenic Escherichia coli (EPEC) it is mediated by several adhesins, including the bundle-forming pilus (BFP) and the EspA filament. Here we report that EPEC forms biofilms on plastic under static conditions and a flowthrough continuous culture system. The abilities of several EPEC isogenic mutants to form biofilms were assessed. Adhesins such as BFP and EspA, important in microcolony formation on epithelial cells, are also involved in bacterial aggregation during biofilm formation on abiotic surfaces. Mutants that do not express BFP or EspA form more-diffuse biofilms than does the wild type. We also determined, using gfp transcriptional fusions, that, consistent with the role of these adhesins in biofilms, the genes encoding BFP and EspA are expressed during biofilm formation. Finally, expression of espA is controlled by a quorum-sensing (QS) regulatory mechanism, and the EPEC qseA QS mutant also forms altered biofilms, suggesting that this signaling mechanism plays an important role in EPEC biofilm development. Taken together, these studies allowed us to propose a model of EPEC biofilm formation.

Biofilms↗

In vitro bactericidal activity of peak serum concentrations of co-amoxiclav, amoxicillin and vancomycin against methicillin-resistant Staphylococcus aureus.

In order to explore the bactericidal activity of concentrations similar to the peak serum concentrations obtained after a single i.v. dose of 2,000/200 mg co-amoxiclav and 500 mg vancomycin, killing curves with co-amoxiclav (69/10 microg/ml), amoxicillin (69 microg/ml), clavulanic acid (10 microg/ml), and vancomycin (15 microg/ml) were performed against two isogenic (ss-lactamase positive and negative) methicillin-resistant Staphylococcus aureus strains in cation-supplemented Mueller-Hinton broth with 2% NaCl incubated at 35 degrees C. Colony counts were performed at 0, 1, 2, 3 and 4 hours in Mueller- Hinton plates supplemented with 4% NaCl and 25 microg/ml oxacillin to measure the resistant population. Similar initial inocula reductions were obtained for amoxicillin-clavulanic acid and vancomycin for both strains, and significant differences were found in comparison to the control. Clavulanic acid decreased the growth rate of the ss-lactamase negative strain when compared to control curves. The penicillin-binding protein 2a affinity of old ss-lactams in conjunction with clavulanic acid overcoming ss-lactamase resistance may explain these results.

Amoxicillin↗